Dept of Pathology

DEPT OF PATHOLOGY

Researcher : Au WH

List of Research Outputs

Liao X., Siu K.Y., Au W.H., Wong E.S.Y., Ngan H.Y.S. and Cheung A.N.Y., Aberrant Activation of Hedgehog Signaling Pathway in Ovarian Cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Researcher : Beh SL

Project Title:	The attitudes of doctors toward rape victims
Investigator(s):	Beh SL
Department:	Pathology
Source(s) of Funding:	Other Funding Scheme
Start Date:	06/1993

Abstract:

To have an objective assessment of what the prevalent attitudes are; to ascertain if there is a gender bias; to compare local data with data from other countries; to compare data with other professional groups.

Project Title:	A STUDY OF ANTECEDENT CLINICAL ENCOUNTERS OF VICTIMS OF DOMESTIC HOMICIDES IN HONG KONG
Investigator(s):	Beh SL
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	01/2006

Abstract:

Prevention of domestic violence is one of the central tasks of family protection. In Hong Kong various tools and guidelines such as the Procedural Guidelines for Handling Battered Spouse Cases (Working group on combating violence, 2004), are researched and published to help social workers and other helping professionals to detect signs of abuse in family. Central monitoring systems, such as the Child Protection Registry and the Central Information System on Battered Spouses Cases and Sexual Violence Cases are established to record and determine the nature of suspected abuse cases. However, few efforts are paid to the understanding of domestic homicides, despite its extreme and grievous nature. The fact that fatal abuse cases were not included in the Child Protection Registry highlights well the lack of understanding and study of lethal cases (Lee& So,2005). This proposal argues that understanding domestic homicides is crucial to family protection, considering the loss of lives, the psychological damage to the surviving family and the psychological and economic impact on the whole society (Brand, Sam, Price,and Richard, 2000). Hong Kong also has a comparatively high proportion of domestic homicides amongst all homicides compared with other jurisdictions, such as the USA and Taiwan (see Appendix 1). The proportion of domestic homicides (DH) in all homicides in Hong Kong has also shown a continuous increase since the 1990s, rising from 24% of all homicides in 1990 to 41% in 2001 (see Appendix 2).One possible reason for the lack of attention to domestic homicides is that domestic homicide is widely regarded as the result of chronic abuse, or as progressing from non-lethal abuse (Browne, 1987; Goetting, 1988; Jurik and Winn, 1990; Stout, 1991; Walker, 1984). However, preliminary research findings of domestic homicides in Hong Kong between 1989 and 2001 suggests that domestic homicides cannot be adequately explained by lethal escalation of chronic abuse. In Hong Kong only 37% of all domestic homicide offenders had a prior history of using violence. This is especially evident in filicides (killing of children by parents) where only a quarter of filicide offenders had prior history of using violence and 42% were driven by non-hostile motives against the victims. Although the history of violence may be the result of under-reporting; its prevalence is unclear and these findings justify further investigations. In particular, it points to the fact that existing risk assessment tools which rely heavily on abusive history and violent records might not be as effective as they are presumed to be in predicting domestic homicides. Theoretically, the differences between lethal and non-lethal cases and the risk of victimization is intriguing. It has been widely reported that the risk of a child being murdered by his or her parents declined as the child grew older in western jurisdictions (Daly and Wilson, 1998). This pattern is also found in Hong Kong. Yet, about 68% to 83% of abused children were older than 6 years old and the highest rate of severe violence occurred around age 8 for boys and age 3 for girls (HK Council of Social Service, 1976; Tang, 1998). Most study on domestic violence in HK determined the prevalence of domestic violence and the demographics of the offenders and victims (Tang, 1998, 1999). This proposal argues that family violence, lethal or non-lethal, is the product of dynamic interactions within the family systems and it should be regarded as a process rather than a static event (Felson & Tedeschi, 1995), usually escalating from minor abuse to more severe attacks (Mouzos, 2000; Dutton, 1998). Most of the domestic homicide victims (85% in one study) had called for police intervention at least once in the 2 years before the homicides (Breedlove, 1977). It is, therefore, crucial to trace the trajectory of domestic violence and homicides for early intervention, and factors which set these two apart.Domestic violence and homicides as a public health issue has gained increasing attention since the last decade and the role of health-care and medical professionals in the prevention of domestic violence gaining more prominence. Domestic violence victims were found to be more vulnerable to a wide array of health problems, especially chronic stress-related problems, such as functional gastrointestinal disorders, appetite loss, viral infections and sexually transmitted diseases (Campbell et al. 2002). They were also more likely to suffer from frequent headaches, migraines, chronic pain syndromes, heart and blood pressures issues symptoms, arthritis, drug and alcohol intoxication, other harm and self-harm behaviors and eating disorders (Wingood, DiClemente and Raj, 2000; Ackard, Neumark-Sztainer, 2002). Research on medical records showed that a considerable proportion of domestic homicide victims had visited Emergency Departments within 2 years prior to their demise and almost all victims who had used the Emergency Department had presented with injuries on at least one encounter. They were also more likely to have injuries on head, neck, chest, abdomen, and perineum (Wadman, et al 1999). Although not necessarily killed by family members, a study on all homicide victims also reported that homicide victims paid more visits to the Emergency Department than the controls, and the frequency increased as the day of the homicide event approached (Cameron S. Crandall, et al. 2004). In view of the limited understanding of domestic homicides, in particular the killings of elderly and other family members, the research team proposes the first victim-based study of domestic homicides in Hong Kong.The objectives are:1) to determine the epidemiology of domestic homicides, offender-victim relationship, demographics, antecedents, situational factors and post-event behaviors;2) to trace the trajectory of domestic homicides as manifested in health problems, such as drug and alcohol intoxication, physical injuries, self-harm behaviors and other health hazards;3) to understand the demographics, nature and frequency of visits of A & E users who died from domestic homicides and determine differences as compared with controls; 4) to increase the alertness of public health professional and facilitate early identification of potential victims who had sought help from the public health systems.

Project Title:	The role of LMO2 in prostate cancer progression
Investigator(s):	Beh SL, Chan KW
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2006

Abstract:

LMO2 belongs to the family of LIM-only proteins (LMO). The family consists of 4 members, designated LMO1-LMO4, known to be important regulators in determining cell fate and controlling cell growth and differentiation. To date, LMO2 has been extensively implicated to play an important role in T-cell leukemia (T-ALL), yet the gene has not been reported to be involved in any solid tumor.Our preliminary data by semiquantitative RT-PCR analysis revealed that mRNA of LMO2 is highly expressed in the more aggressive, androgen-independent PC3 and DU145 prostate cancer cell lines but not in the less aggressive, androgen-dependent LNCaP cell line. The aim of this proposed project is to further study the role LMO2 plays in driving prostate cancer progression.We aim to (1) determine the protein expression of LMO2 in various prostate cancer cell lines (PC3, DU145, LNCaP and immortalized normal prostate HPr-1) by western blotting, as a means to confirm our RT-PCR data; (2) stably transfect the LMO2 gene to LNCaP cell line, which was found to have low expression of LMO2; (3) silence the LMO2 gene by siRNA technique in DU145 cell line, which was found to have high expression of LMO2; (4) study the effects of varying LMO2 expressions on invasiveness of prostate cancer cell lines by in vitro assays such as wound healing, and matrigel invasion; (5) determine the expression of LMO2 in a prostate tissue microarray, incorporating many prostate cancer samples, recently constructed in our laboratory, by immunohistochemistry; (6) correlate LMO2 expression in prostate cancer with the patients' clinico-pathological parameters; and (7) determine the mechanistic involvement of LMO2 in the molecular network of prostate cancer progression.

List of Research Outputs

Beh S.L., Member of International Editorial Board, In: Ryoji Sakai, Journal Of Medical Safety. Union of Risk Management for Preventive Medicine, 2008.

Beh S.L., Sudden Cardiac Death, 24th World Congress Of Pathology And Laboratory Medicine. 2007.

Beh S.L., Why Are We Investigating Death, 9th Indo-pacific Congress On Legal Medicine And Forensic Sciences Of The Indo-pacific Association Of Law, Medicine And Science. 2007.

Wong P.W.C., Chan W.S.C. and Beh S.L., What can we do to help and understand survivors of suicide in Hong Kong? , Crisis. Hogrefe, 2007, 28: 183-189.

Researcher : Cao X

List of Research Outputs

Lam Q.L.K., Zheng B., Jin D., Cao X. and Lu L., Leptin induces CD40 expression through the activation of AKT in murine dendritic cells, J Biol Chem. 2007, 38: 27587-27597.

Researcher : Chan ASY

List of Research Outputs

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Kosinski C., Li V.S.W., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors, Proc Natl Acad Sci U S A. 2007, 104(39): 15418-23.

Li V.S.W., Kosinski C., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression profiling of human colon top versus basal crypts and identification of BMP antagonists as candidates for intestinal stem cell niche, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Suehiro Y., Wong C.W., Chirieac L.R., Kondo Y., Shen L., Webb C.R., Chan Y.W., Chan A.S.Y., Chan T.L., Wu T.T., Rashid A., Hamanaka Y., Hinoda Y., Shannon R.L., Morris J., Issa J.P., Yuen S.T., Leung S.Y. and Hamilton S.R., Epigenetic-Genetic Interactions in the APC/WNT, RAS/RAF, and P53 Pathways in Colorectal Carcinoma, Clin Cancer Res. 2008, 14(9): 2560-9.

Researcher : Chan DW

Project Title:	Characterization of the roles of Dual specificity MAPK phosphatase 3 (MKP-3) in ovarian cancer
Investigator(s):	Chan DW, Ngan HYS
Department:	Obstetrics & Gynaecology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2006

Abstract:

Ovarian cancer is one of the leading causes of death in women. Although there have been advances in the treatment of ovarian cancer, the associated mortality rate of this cancer has not improved significantly over the past decade. Therefore, understanding the molecular mechanisms in the development of ovarian cancer through identification and characterization of oncogenes and tumor suppressor genes will help discovery of novel targets for therapies. To achieve this, we have previously done cDNA microarray analysis on ovarian cancer cell lines and immortalized human normal surface epithelial cell lines (HOSEs). We found that a gene called Dual specificity MAPK phophatase 3 (MKP-3/DUSP6) was downregulated in ovarian cancer cells. Many studies have documented that MKP-3 possesses anti-tumorigenic effect on pancreatic cancer through inactivation of ERK activity (Furukawa et al., 1998; Furukawa et al., 2003). The finding of underexpression of MKP-3 in ovarian cancer cells indicates that this gene may play a role in the development of ovarian cancer. The objectives of this research proposal are: 1. To evaluate the expression status of MKP-3 in ovarian cancer cell lines, HOSEs, and clinical samples. Clinicopathological correlation will be analyzed with the expression status of MKP-3 in patients’ samples. 2. To investigate the relationship between MKP-3 expression status and ERK1/2 activity in ovarian cancer cell lines and clinical samples. 3. To evaluate the change of tumorigenicity of ovarian cancer cells by stably transfected with MKP-3 expressing constructs and/or knock-down MKP-3 by RNAi technique. 4. To delineate the signalling pathways in MKP-3/ERK/downstream targets regulation in ovarian cancer cells.

Researcher : Chan EYT

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Ho M.H.K., Chan E.Y.T., Lee T.L. and Lau Y.L., The application of allergy tests in childhood allergy, Hong Kong Journal of Dermatology Venerology. 2008, 16: 77-91.

Researcher : Chan GSW

List of Research Outputs

Chan G.S.W., Lam M.F., Au W.Y., Chim S., Tse K.C., Lo S.H., Fung S.H., Lai K.N. and Chan K.W., Clinicopathologic analysis of renal biopsies after haematopoietic stem cell transplantation, Nephrology. 2008, 13(4): 322-330.

Chan G.S.W., Lam M.F., Au W.Y., Tse K.C., Chim S., Fung S.H., Lo S.H.K., Lai K.N. and Chan K.W., Renal Pathology after Haematopoietic Stem Cell Transplantation (HSCT): A study of 13 patients, 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Kuala Lumpur, Malaysia. 2007.

Chan K.W., Chan G.S.W., Tse K.C. and Chan C.P., Genotypic and phenotypic divergence: lessons from a family of Fabry’s disease, 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Kuala Lumpur, Malaysia. 2007.

Chan K.W. and Chan G.S.W., Immunohistochemical markers, are we having too many choices?, 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Kuala Lumpur, Malaysia, 2007 . 2007.

Chan K.W. and Chan G.S.W., The Occurrence of Polyomavirus (BK Virus and its Variants) Transplant Nephropathy in Hong Kong and Macau, 11th Asian Pacific Congress of Nephrology, 5 - 8 May 2008, Kuala Lumpur Convention Centre, Malaysia. 2008.

Researcher : Chan HY

List of Research Outputs

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Siu K.Y., Chan H.Y., Kong S.H., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Differential expression of folate receptor alpha and reduced folate carrier and effect of folate in ovarian cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Siu K.Y., Woo N.W., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th Annual Growth Factor and Signal Transduction Symposium titled "Senescence, Aging and Cancer",Iowa, 26-29 July . 2007.

Woo N.W.S., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th annual Growth Factor and Signal Transduction Symposium, Iowa, 26-29 July. 2007.

Researcher : Chan KK

List of Research Outputs

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Yuen H.F., Chan Y.P., Chan K.K., Chu Y.Y., Wong M.L.Y., Law S.Y.K., Srivastava G., Wong Y.C., Wang X. and Chan K.W., Id-1 and Id-2 are markers for metastasis and prognosis in oesophageal squamous cell carcinoma, Br J Cancer. 2007, 97(10): 1409-15.

Researcher : Chan KW

Project Title:	Prognostic significance of Id proteins in esophageal squamous cell carcinoma (ESCC)
Investigator(s):	Chan KW
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2006

Abstract:

Purpose of the proposed project: The proposed project aims to clarify the roles of Id-proteins in the oncogenesis and progression of esophageal squamous cell carcinoma (ESCC) and to explore the potentials for using them as prognostic markers and targets for therapy of ESCC. To achieve the goals, the following two approaches are to be adopted. (i) To study how Id proteins expressions differ in normal human esophageal epithelial and esophageal cancer cell lines. (ii) To establish the prognostic significance of the expression of each Id proteins in ESCC by determining and correlating their expression levels in primary ESCC with clinical staging, histological grading, disease progression and clinical outcome. Background: Esophageal cancer is the eighth most common cancer and the sixth most common cancer-related death worldwide. A 20-fold higher risk was observed in China when compared to low risk areas (Parkin et al., 2005). Therefore, a good understanding of the molecular biology and the identification of prognostic markers are of particular importance to the local community. ESCC, the most common histological subtype of esophageal cancer, is associated with high morbidity and mortality (Lam, 2000). Recent studies have demonstrated that the expression of several proteins in ESCC, such as mannose-binding lectin-associated serine protease-2 (Verma et al., 2006), cyclooxygenase-2 (Zhi et la., 2006) and Fas (Chan et al., 2006), may have useful prognostic value. Carcinogenesis and progression of ESCC are multifactorial. Identification of new prognostic markers in ESCC can improve the clinical management on ESCC patients by selecting groups of patients with higher risks of progression for more aggressive therapy. By understanding the detailed molecular pathogenesis of these potential new prognostic markers, the information can be exploited for development of effective, targeted therapeutic intervention (Fong et al., 2004) . There are four known members of the "inhibitors of differentiation" or "inhibitors of DNA-binding" (Id-1, Id-2, Id-3 and Id-4) subfamily of helix-loop-helix genes. Id proteins have been shown to be involved in many important cellular processes as well as carcinogenesis, including cell cycle control (Zebedee and Hara, 2001), cancer development (Lasorella et al., 2001), angiogenesis (Benezra et al., 2001) and cell survival (Wong et al., 2004). Although they are in the same family, their roles in carcinogenesis in different types of cancer are different. The clinical significance of these Id proteins has recently been studied in hepatocellular carcinoma (Damdinsuren et al., 2005), cervical cancer (Schindl et al., 2001), colorectal cancer (Umetani et al., 2004), gastric cancer (Jang et al., 2005) and breast cancer (Stighall et al., 2005). By immunohistochemistry, we have shown that the expression of Id proteins is dysregulated in prostate cancer while Id proteins are differentially expressed during cancer progression (Yuen et al., 2006). Id-1 expression is significantly increased in primary ESCC compared to normal esophageal epithelium (Hu et al., 2001). Id proteins are likely to play important but variable roles in the transformation and progression of different types of cancer and Id proteins are potential prognostic markers in those cancers. The proposed project aims to further clarify the roles of Id-proteins in the oncogenesis and progression of ESCC and to explore their potentials for being prognostic markers and therapeutic targets in this cancer. References: Benezra R, Rafii S, Lyden D. The Id proteins and angiogenesis. Oncogene. 2001 20:8334-8341. Chan KW, Lee PY, Lam AK, Law S, Wong J, Srivastava G. Clinical relevance of Fas expression in oesophageal squamous cell carcinoma. J Clin Pathol. 2006 59:101-104. Damdinsuren B, Nagano H, Kondo M, Yamamoto H, Hiraoka N, Yamamoto T, Marubashi S, Miyamoto A, Umeshita K, Dono K, Nakamori S, Wakasa K, Sakon M, Monden M. Expression of Id proteins in human hepatocellular carcinoma: relevance to tumour dedifferentiation. Int J Oncol. 2005 26:319-327. Fong S, Debs RJ, Desprez PY. Id genes and proteins as promising targets in cancer therapy. Trends Mol Med. 2004 10:387-392. Hu YC, Lam KY, Law S, Wong J, Srivastava G. Identification of differentially expressed genes in esophageal squamous cell carcinoma (ESCC) by cDNA expression array: overexpression of Fra-1, Neogenin, Id-1, and CDC25B genes in ESCC. Clin Cancer Res. 2001 7:2213-2221. Jang TJ, Jung KH, Choi EA. Id-1 gene downregulation by sulindac sulfide and its upregulation during tumour development in gastric cancer. Int J Cancer. 2006 118:1356-1363. Lam AK. Molecular biology of esophageal squamous cell carcinoma. Crit Rev Oncol Hematol. 2000 33:71-90. Lasorella A, Uo T, Iavarone A. Id proteins at the cross-road of development and cancer. Oncogene. 2001 20:8326-8333. Parkin DM, Bray F, Ferlay J, Pisani P. Global cancer statistics, 2002. CA Cancer J Clin. 2005 55:74-108. Schindl M, Oberhuber G, Obermair A, Schoppmann SF, Karner B, Birner P. Overexpression of Id-1 protein is a marker for unfavorable prognosis in early-stage cervical cancer. Cancer Res. 2001 61:5703-5736. Stighall M, Manetopoulos C, Axelson H, Landberg G. High ID2 protein expression correlates with a favourable prognosis in patients with primary breast cancer and reduces cellular invasiveness of breast cancer cells. Int J Cancer. 2005 115:403-411. Wong YC, Wang X, Ling MT. Id-1 expression and cell survival. Apoptosis. 2004 9:279-289. Umetani N, Takeuchi H, Fujimoto A, Shinozaki M, Bilchik AJ, Hoon DS. Epigenetic inactivation of ID4 in colorectal carcinomas correlates with poor differentiation and unfavorable prognosis. Clin Cancer Res. 2004 10:7475-7483. Verma A, Matta A, Shukla NK, Deo SV, Gupta SD, Ralhan R. Clinical significance of mannose-binding lectin-associated serine protease-2 expression in esophageal squamous cell carcinoma. Int J Cancer. 2006 118:2930-2935. Yuen HF, Chua CW, Chan YP, Wong YC, Wang X, Chan KW. Id proteins expression in prostate cancer: high-level expression of Id-4 in primary prostate cancer is associated with development of metastases. Mod Pathol. 2006 19:931-941. Zebedee Z, Hara E. Id proteins in cell cycle control and cellular senescence. Oncogene. 2001 20:8317-8325. Zhi H, Wang L, Zhang J, Zhou C, Ding F, Luo A, Wu M, Zhan Q, Liu Z. Significance of COX-2 expression in human esophageal squamous cell carcinoma. Carcinogenesis. 2006 27:1214-1221.

List of Research Outputs

Chan D.T.M., Lin A.W., Tang S.C.W., Qian J.Q., Lam M.F., Ho Y.W., Tse K.C., Chan K.W., Lai K.N. and Tang C.S.O., Prospective controlled study on mycophenolate mofetil and prednisolone in the treatment of membranous nephropathy with nephrotic syndrome, Nephrology. Australia, Blackwell Publishing, 2007, 12: 576-581.

Chan G.S.W., Lam M.F., Au W.Y., Chim S., Tse K.C., Lo S.H., Fung S.H., Lai K.N. and Chan K.W., Clinicopathologic analysis of renal biopsies after haematopoietic stem cell transplantation, Nephrology. 2008, 13(4): 322-330.

Chan G.S.W., Lam M.F., Au W.Y., Tse K.C., Chim S., Fung S.H., Lo S.H.K., Lai K.N. and Chan K.W., Renal Pathology after Haematopoietic Stem Cell Transplantation (HSCT): A study of 13 patients, 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Kuala Lumpur, Malaysia. 2007.

Chan K.W., Chan G.S.W., Tse K.C. and Chan C.P., Genotypic and phenotypic divergence: lessons from a family of Fabry’s disease, 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Kuala Lumpur, Malaysia. 2007.

Chan K.W. and Chan G.S.W., Immunohistochemical markers, are we having too many choices?, 24th World Congress of Pathology and Laboratory Medicine, 20-24 August 2007, Kuala Lumpur, Malaysia, 2007 . 2007.

Chan K.W. and Chan G.S.W., The Occurrence of Polyomavirus (BK Virus and its Variants) Transplant Nephropathy in Hong Kong and Macau, 11th Asian Pacific Congress of Nephrology, 5 - 8 May 2008, Kuala Lumpur Convention Centre, Malaysia. 2008.

Chung Y.M.F., Lam A.K.Y., Luk J.M.C., Law S.Y.K., Chan K.W., Lee P.Y. and Wong J., Altered E-cadherin expression and p120 catenin localization in esophageal squamous cell carcinoma, Annals of Surgical Oncology. 2007, 14(11): 3260-3267.

Dai Y., Qiao L., Chan K.W., Zou B., Ma J., Lan H.Y., Gu Q., Li Z., Wang Y., Wong B.L.W. and Wong B.C.Y., Loss of XIAP sensitizes Rosiglitazone-induced growth inhibition of colon cancer in vivo, Int J Cancer. 2008, 122: 2858-63.

Howard E.W., Leung C.L., Yuen H.F., Chua C.W., Lee D.T.W., Chan K.W., Wang X. and Wong Y.C., Decreased adhesiveness, resistance to anoikis and suppression of GRP94 are integral to the survival of circulating tumor cells in prostate cancer, Clinical and Experimental Metastasis. 2008, 25: 497-508.

Hui K.C., Chan K.W., Luk J.M.C. and Law S.Y.K., The roles of survivin in esophageal squamous cell carcinoma, The 12th Research Postgraduate Symposium, The University of Hong Kong, Li Ka Shing Faculty of Medicine, Hong Kong, 12 - 14 December 2008. 2007.

Lui S.L., Tsang R.C.W., Zhang Q., Chan K.W., Yung S.S.Y. and Chan D.T.M., Rapamycin attenuates the severity of established nephritis in NZB/W F1 mice, Journal of the American Society of Nephrology. 2007, 18: 195A.

Lui S.L., Tsang R.C.W., Chan K.W., Zhang Q., Tam S., Yung S.S.Y. and Chan D.T.M., Rapamycin attenuates the severity of established nephritis in lupus-prone NZB/W F1 mice, Nephrology Dialysis Transplantation. 2008, 23: 2768-2776.

Lui S.L., Yung S.S.Y., Tsang R.C.W., Zhang Q., Chan K.W., Tam S. and Chan D.T.M., Rapamycin prevents the development of nephritis in lupus-prone NZB/W F1 mice, Lupus. 2008, 17: 305-313.

Ma S.K.Y., Lee K.Y., Zheng B. and Chan K.W., CD133(+) HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway., Oncogene. 2008, 27: 1749-1758.

Ma S.K.Y., Lee K.W., Zheng B., Chan K.W. and Guan X.Y., CD133+ HCC cancer stem cells promote chemoresistance by preferential expression of the Akt/PKB survival pathway, Oncogene. 2008, 27: 1749-1758.

Ma S.K.Y., Guan X.Y., Lee K.W. and Chan K.W., Clinicopathological significance of missing in metastasis B expression in hepatocellular carcinoma, Hum Pathol. 2007, 38(8): 1201-6.

Ma S.K.Y., Hu L., Huang X.H., Cao L.Q., Chan K.W., Wang Q. and Guan X.Y., Establishment and characterization of a human cholangiocarcinoma cell line., Oncology Report. 2007, 18: 1195-1200.

Qiao L., Dai Y., Gu Q., Chan K.W., Zou B., Ma J., Wang J., Pang R.W.C., Lan H.Y. and Wong B.C.Y., Down-regulation of X-linked inhibitor of apoptosis synergistically enhanced peroxisome proliferator-activated receptor {gamma} ligand-induced growth inhibition in colon cancer, Mol Cancer Ther. 2008, 7: 2203-2211.

Qiao L., Dai Y., Gu Q., Chan K.W., Ma J., Lan H.Y., Zou B., Rocken C , Ebert MP and Wong B.C.Y., Loss of XIAP sensitizes colon cancer cells to PPARgamma independent antitumor effects of troglitazone and 15-PGJ2. , Cancer Lett. 2008, 268: 260-71.

Yuen H.F., Chan Y.P., Chan K.K., Chu Y.Y., Wong M.L.Y., Law S.Y.K., Srivastava G., Wong Y.C., Wang X. and Chan K.W., Id-1 and Id-2 are markers for metastasis and prognosis in oesophageal squamous cell carcinoma, Br J Cancer. 2007, 97(10): 1409-15.

Zheng B., Chan K.W., Lin Y., Zhao G., Chan C.S., Zhang H., Chen H., Wong S.S.Y., Lau S.K.P., Woo P.C.Y., Chan K.H., Jin D. and Yuen K.Y., Delayed antiviral plus immunomodulator treatment still reduces mortality in mice infected by high inoculum of influenza A/H5N1 virus., Proc Natl Acad Sci USA.. 2008, 105(23): 8091-8096.

Researcher : Chan KYQ

List of Research Outputs

Ip P.P.C., Lam K.W., Cheung C.L., Yeung C.W., Pun T.C., Chan K.Y.Q. and Cheung A.N.Y., Tranexamic Acid-associated Necrosis and Intralesional Thrombosis of Uterine Leiomyomas: A Clinicopathologic Study of 147 Cases Emphasizing the Importance of Drug-induced Necrosis and Early Infarcts in Leiomyomas , American Journal of Surgical Pathology. 2007, 31(8): 1215-1224.

Siu K.Y., Chan H.Y., Kong S.H., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Differential expression of folate receptor alpha and reduced folate carrier and effect of folate in ovarian cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Siu K.Y., Woo N.W., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th Annual Growth Factor and Signal Transduction Symposium titled "Senescence, Aging and Cancer",Iowa, 26-29 July . 2007.

Woo N.W.S., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th annual Growth Factor and Signal Transduction Symposium, Iowa, 26-29 July. 2007.

Zhang H., Siu K.Y., Li A.S.M., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Researcher : Chan LC

Project Title:	Ras signaling in human leukaemia
Investigator(s):	Chan LC, Kong CT
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2005

Abstract:

To study: (1) Identification of the possible interaction between other MLL fusion partners and components of Ras-mediated pathways. i) design of MLL fusion partner constructs. ii) analysis of transactivation of E1K-1, a target of Ras pathway signaling. iii)soft agar transformation assay. (2) Molecular dissection of the potential functional link between Ras-signaling and MLL-AF6 fusion mediated leukaemogenesis using the ML-1 cell line. i) molecular analysis of Ras signaling pathway following suppression of MLL-AF6 expression in ML-1 cell line . ii) effects of inhibition of Ras signaling pathway on cellular growth and apotosis of ML-1 cell line . (3) Validation of the pathogenic significance of Ras-signaling pathway in MLL fusion mediated leukaemogenesis using an Mll-Een knock-in model. i) effects on suppression of Ras signaling pathway on growth and differentiation of embryonic bodies and haemopoiesis colonies derived from Mll-Een targeted ES cells.

Project Title:	ATM mutations in childhood leukaemia
Investigator(s):	Chan LC, Ha SY
Department:	Pathology
Source(s) of Funding:	Michael Kadoorie Cancer Genetics Research Fund
Start Date:	01/2005

Abstract:

To screen for ATM gene mutations in childhood ALL samples from Hong Kong, Shanghai, Tokyo and London, UK; to determine the germline status of the ATM gene in patients whose diagnostic samples showed mutations; to determine the presence of ATM mutations or allelic variations in the normal population of samples.

Project Title:	Expression and cellular localization studies of TCF12, a new leukaemia fusion partner gene, in normal and malignant haemopoiesis and the generation of mouse embryonic stem cells bearing MLL-TCF12 through homologous recombination
Investigator(s):	Chan LC
Department:	Pathology
Source(s) of Funding:	Germany/Hong Kong Joint Research Scheme
Start Date:	01/2007

Abstract:

1. To determine the expression profile of TCF12 in normal and malignant haemopoietic cells 2. To determine the cellular localization of TCF12 3. To generate through homologous recombination targeted mouse embryonic stem cells with MLL-TCF12

Project Title:	Application of the FICTION (fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms) technique on blood and bone marrow aspirate specimens to study leukaemias
Investigator(s):	Chan LC
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	01/2007

Abstract:

Fluorescence in-situ hybridization (FISH) is an established method to detect numerical and structural chromosomal abnormalities in leukaemia. Using fluorescent dye-labelled nucleic acid probes, genetic defects including specific chromosome loss/gain, deletion and translocation can be detected in interphase leukaemic cells, thus obviating the need to perform cell culture to analyse cells in mitosis as in conventional cytogenetics. One distinct disadvantage of FISH is inability to correlate cell phenotype with genetic defects studied. In solid tumours this is less of a problem because of their cohesive and relative monotonous nature, thus rendering identification of tumour cells easier. However, the cell types in leukaemias are very heterogeneous and many non-neoplastic cells are often admixed with leukaemic cells in blood and bone marrow aspirate. Prior purification by cell sorting, using either flow or magnetic activated technique, is necessary if phenotype-genotype correlation is indicated. Sorting is also required to minimize false negativity due to the presence of contaminating normal cells. FICTION protocols for histological sections including trephine biopsy have been published. However, a corresponding technique for blood and marrow aspirate specimens has not been well studied. The objective of the research proposal is to develop a standard FICTION technique applicable to blood and bone marrow aspirate specimens. References: Weber-Matthiesen K, Winkemann M, Muller-Hermelink A, Schlegelberger B, Grote W. Simultaneous detection of the immunophenotypic markers and genetic aberrations on routinely processed paraffin sections of lymphoma samples by means of the FICTION technique. J Histochem Cytochem. 1992;40:171-5. Martinez-Ramirez A, Cigudosa JC, Maestre L, Rodriguez-Perales S, Haralambieva E, Benitez J, Roncador G. Simultaneous detection of the immunophenotypic markers and genetic aberrations on routinely processed paraffin sections of lymphoma samples by means of the FICTION technique. Leukemia. 2004;18:348-53. Korac P, Jones M, Dominis M, Kusec R, Mason DY, Banham AH, Ventura RA. Application of the FICTION technique for the simultaneous detection of immunophenotype and chromosomal abnormalities in routinely fixed, paraffin wax embedded bone marrow trephines. J Clin Pathol. 2005;58:1336-8.

List of Research Outputs

Chan L.C., American Journal of Haematology. 2007.

Chan L.C., Chronic myeloproliferative disorders – perspectives from FISH and molecular diagnosis, Department of Pathololgy, Korean University Guro Hospital. Seoul, South Korea. 2008.

Chan L.C., Plenary speaker on the topic “An introduction to undergraduate medial education and PBL”, Annual Spring Meeting, Korean Society of Pathologists. Seoul, South Korea. 14-16 May. 2008.

Chan L.C., Speaker on the Session “A Decade of Curriculum Reform” - Topics: PBL: Behind the Buzz-word”, Frontiers in Medical Education HKU 2007, Faculty of Medicine, The University of Hong Kong. 2007.

Chan L.C., Teaching and Learning of Haematology – integrating basic and clinical science through PBL, Korea University College of Medicine. 2008.

Chan L.C., The role of a PBL tutor: A personal perspective, 2007 International Problem-Based Learning Workshop, Kaohsiung Medical University, Taiwan. The Kaohsiung Journal of Medical Sciences. 2007, Vol 24, No. 3: Suppl, S34-S38.

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Chen Y.X., Man K., Ling G.S., Chen Y., Sun B., Cheng Q., Wong O.H., Lo C.K., Ng I.O.L., Chan L.C., Lau G., Lin S.C.L., Huang F. and Huang F.P., A crucial role for dendritic cell (DC) IL-10 in inhibiting successful DC-based immunotherapy: superior antitumor immunity against hepatocellular carcinoma evoked by DC devoid of IL-10, Journal of Immunology. 2007, 179(9): 6009-6015.

Cheung N., Chan L.C., Thompson A., Cleary M.L. and So C.W., Protein arginine-methyltransferase-dependent oncogenesis, Nat Cell Biol. 2007, 9(10): 1208-15.

Gibney G.T., Panhuysen C.I.M., So J.C.C., Ma E.S.K., Ha S.Y., Li C.K., Lee A.C.W., Li C.K., Yuen H.L., Lau Y.L., Johnson D.M., Farrell J.J., Bisbee A.B., Farrer L.A., Steinberg M.H., Chan L.C. and Chui D.H.K., Variation and heritability of Hb F and F-cells among beta-thalassemia heterozygotes in Hong Kong., Am J Hematol. 2008, 458-464.

Lee A.C., Ma E.S.K., Chan A.Y., Szeto S.C. and Chan L.C., Double heterozygosity for Hb New York [beta 113 GTG-->GAG; VAL-->GLU] and beta degrees-thalassemia mutations manifests as a thalassemia trait, Pediatr Hematol Oncol. 2008, 25(3): 227-31.

So J.C.C., Wan T.S.K., Yip S.F. and Chan L.C., A dual colour dual fusion fluorescence in situ hybridisation study on the genesis of complex variant translocations in chronic myelogenous leukaemia, Oncol Rep. 2008, 19(5): 1181-1184.

So J.C.C., Ma E.S.K., Wan T.S.K., Yip S.F. and Chan L.C., Clinicopathological features of unbalanced translocation Der(1;7)(q10;p10) in myeloid neoplasms, Leuk Res. 2008, 32(6): 1000-1001.

Sun Q., Kong C.T., Huang F. and Chan L.C., Aberrant Dendritic Cell Differentiation Initiated by Mll-Een Fusion Gene Does Not Requre Leukaemic Transformation., Journal of Leukocyte Biology. 2008, 83 (1): 173 - 180.

Researcher : Chan LK

List of Research Outputs

Yam J.W.P., Tse Y.T., Ko C.F., Chan L.K., Sze M.F. and Ng I.O.L., Tensin2 with growth suppression and apoptosis induction activities is underexpressed in hepatocellular carcinoma, 2008 Proceedings of the American Association for Cancer Research, San Diego, CA, USA, April. 2008.

Researcher : Chan SK

List of Research Outputs

Yuan X.J., Chan G.C.F., Chan S.K., Shek T.W.H., Kwong D.L.W., Wei W.I., Ha S.Y. and Chiang A.K.S., Treatment outcome of rhabdomyosarcoma in Hong Kong Chinese children, Hong Kong Medical Journal. 2008, 14(2): 116-123.

Researcher : Chan TL

Project Title:	Molecular characterisation of the serrated neoplasia pathway and its role in the development of colorectal cancer with mismatch repair deficiency
Investigator(s):	Chan TL, Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	01/2005

Abstract:

Colorectal cancer (CRC) is one of the commonest cancers world-wide with 850,000 new cases each year. Majority of CRC are known to develop through an adenoma-carcinoma sequence, with molecular genetic changes that characterize each transition step. Adenoma is considered pre-malignant and early prevention and treatment of CRC is possible through regular endoscopic surveillance and removal of tumour at the adenoma stage. However, despite regular surveillance, some patients still develop CRC. One of the possibilities is that there exists another pathway of tumour development. Recently, there are data to suggest the existence of an alternative route, the serrated neoplasia pathway, for CRC development. This latter pathway includes those serrated polyps (SP) that span a morphological spectrum from hyperplastic polyp (HP) to serrated adenoma (SA). The earliest member in this pathway, HP, is a very common lesion in aged individuals and it is a long held belief that HPs are innocent with little propensity for malignant progression. However, a small subset of HPs may progress but the criteria to distinguish the high risk versus the low risk ones are unclear. Concurrently, it is known that a subset (15%) of sporadic CRC manifest a form of genetic instability called microsatellite instability (MSI). This is due to promoter methylation leading to loss of expression of the DNA mismatch repair protein MLH1. The evolution of this subset of CRC is unclear as a preceding adenoma phase is very rarely seen. Recently, studies by us and others have provided molecular evidences to suggest that there may be a strong link between the serrated neoplasia pathway and the development of sporadic CRC with MSI. These include the identification of a high incidence of BRAF mutation in HPs and SAs, the occurrence of MSI in some SPs and the selective association of BRAF mutation with sporadic late-onset CRC with MSI. Our recent pilot study has shown that MLH1 inactivation can be detected in SPs at its very early phase and these can just involve several crypts within the lesion. With these data, we propose to perform a large scale phenotypic and molecular characterization of serrated polyps at their early phase of evolution to define the link between SPs and MSI CRC, and the temporal sequence of genetic changes in this pathway. Aims: 1. To look for evidence of MLH1 inactivation in various stages of evolution of serrated polyps, and to document its incidence and phenotypic characteristics. 2. To document the occurrence of microsatellite instability and frameshift mutation in growth regulatory genes containing microsatellite encoding region as consequences of MLH1 inactivation in various stages of serrated polyps. 3. To analyse the inter-relationship between MLH1 inactivation, BRAF/KRAS mutations and presence of the CpG Island Methylator Phenotype in SPs and their temporal sequence of occurrence. 4. To look for alteration in major molecular genetic pathways in SPs with MLH1 inactivation. 5. To look for evidence of a field effect of MLH1 inactivation in the colon in patients with sporadic late-onset MSI colorectal cancer with MLH1 promoter methylation. 6. To look for clinical, morphological or molecular markers that can distinguish SPs with high risk of progression to MSI CRC.

Project Title:	Allele-specific imbalance in gene expression as a cause for hereditary colorectal cancer
Investigator(s):	Chan TL, Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Michael Kadoorie Cancer Genetics Research Fund
Start Date:	01/2005

Abstract:

To look for allele-specific imbalance in gene expression of the Adenomatous Polyposis Coli (APC), MSH2 and MLH1 genes in FAP or HNPCC patients with undetectable germline mutation; to confirm the pathological significance of the allele with a lower gene expression level by examination for co-segregation with disease and loss of the wild-type allele in cancer tissue; to look for mechanisms that can account for the reduced gene expression level in the disease allele.

Project Title:	Molecular characterisation of the serrated neoplasia pathway and its role in the development of colorectal cancer with mismatch repair deficiency
Investigator(s):	Chan TL, Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	12/2005

Abstract:

To look for evidence of MLH1 inactivation in various stages of evolution of serrated polyps, and to document its incidence and phenotypic characteristics; to document the occurrence of microsatellite instability and frameshift mutation in growth regulatory genes containing microsatellite encoding region as consequences of MLH1 inactivation in various stages of serrated polyps; to analyse the inter-relationship between MLH1 inactivation, BRAFIKRAS mutations and presence of the CpG Island Methylator Phenotype in SPs and their temporal sequence of occurrence; to look for alteration in major molecular genetic pathways in SPs with MLH1 inactivation; to look for evidence of a field effect of MLH1 inactivation in the colon in patients with sporadic late-onset MSI colorectal cancer with MLH1 promoter methylation; to look for clinical, morphological or molecular markers that can distinguish SPs with high risk of progression to MSI CRC.

Project Title:	Expression of microRNAs in various stages and pathways of colorectal carcinogenesis
Investigator(s):	Chan TL, Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	02/2006

Abstract:

MicroRNAs (miRNAs) are a newly discovered class of non-protein coding RNAs that play important roles in embryonic development and many biological processes in diverse species. They function to regulate expression of other protein-coding genes by promoting their mRNA degradation or inhibiting their protein translation. Though the precise functions of individual miRNA and the repertoire of target genes that they each regulates are mostly unknown, emerging data suggest their involvement in regulating some major oncogenic pathways including those mediated by the RAS and the c-myc genes. miRNAs reside frequently in genomics regions involved in cancers. Deranged expression of miRNAs has been reported in several cancer types. The miRNA expression profile can distinguish histological cell lineage of cancers and can even distinguish molecular pathways of transformation for cancers derived from the same cell lineage. Colorectal cancer (CRC) is one of the most common cancers worldwide. Several major molecular pathways of CRC development is currently known, including the APC/KRAS/p53/chromosome instability pathway and the microsatellite instability/TGF pathway. Currently, little is known regarding the expression profiles of miRNAs in colon, and whether there is deregulated expression of specific miRNAs in various stages or pathways of CRC development. In this study we aim to systematically profile the expression of miRNAs, using a high-throughput quantitative RT-PCR analysis method, in various stages and pathways of CRC development, with an aim to gain a better understanding of their possible involvement in colon carcinogenesis. Aims 1) To profile the expression of miRNAs in normal mucosae, adenomas and carcinomas of the colon, and to compare the differences in miRNA expression profiles of the progenitor cells residing at the base of the crypt and the mature cells residing at the top. 2) To compare the variation in expression of miRNAs in different molecular subtypes of CRC including those with microsatellite instability, microsatellite stable or microsatellite and chromosome stable phenotype. 3) To identify specific miRNAs of interest that show altered expression in the course of CRC development, or in specific molecular subgroups of CRC 4) To investigate the function of these miRNAs by over-expression or gene silencing approach in cell culture system. 5) To delineate the putative protein-coding target(s) regulated by these miRNA(s) using bioinformatics approach and their validation by western blotting.

Project Title:	Expression of microRNAs in various stages and pathways of colorectal
Investigator(s):	Chan TL, Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2007

Abstract:

To profile the expression of miRNAs in normal mucosae, adenomas of the colon, and to compare the differences in miRNA expression profiles of the progenitor cells residing at the base of the crypt and the mature cells residing at the top; to compare the variation in expression of miRNAs in different molecular subtypes of CRC including those with microsatellite instability, microsatellite stabel or microsatellite and chromosome stable phenotype; to identify specific miRNAs if interest that show altered expression in the course of CRC development, or in specific molecular subgroups of CRC; to investigate the function of these miRNAs by over-expression or gene silencing appraoch in cell culture system; to delineated the putative protein-coding target(s) regulated by these miRNA(s) using bioinformatics approach and their validation by western blotting.

List of Research Outputs

Chan T.L., Frontiers in Medical Laboratory Sciences: Molecular Genetic Screening – New Insight of Medical Laboratories in Hong Kong, Annual Scientific Meeting 2007, Hong Kong Institute of Medical Laboratory Sciences Limited. 2007.

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Chan T.L., Mechanism of Carcinogenesis: Genetics and Epigenetics of Colorectal Cancer, 14th Hong Kong International Cancer Congress and 3rd Annual Meeting of Centre for Cancer Research. 2007.

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Kosinski C., Li V.S.W., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors, Proc Natl Acad Sci U S A. 2007, 104(39): 15418-23.

Li V.S.W., Kosinski C., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression profiling of human colon top versus basal crypts and identification of BMP antagonists as candidates for intestinal stem cell niche, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Schetter A.J., Leung S.Y., Sohn J.J., Zanetti K.A., Bowman E.D., Yanaihara N., Yuen S.T., Chan T.L., Kwong D.L., Au G.K., Liu C.G., Calin G.A., Croce C.M. and Harris C.C., MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma, JAMA. 2008, 299(4): 425-36.

Suehiro Y., Wong C.W., Chirieac L.R., Kondo Y., Shen L., Webb C.R., Chan Y.W., Chan A.S.Y., Chan T.L., Wu T.T., Rashid A., Hamanaka Y., Hinoda Y., Shannon R.L., Morris J., Issa J.P., Yuen S.T., Leung S.Y. and Hamilton S.R., Epigenetic-Genetic Interactions in the APC/WNT, RAS/RAF, and P53 Pathways in Colorectal Carcinoma, Clin Cancer Res. 2008, 14(9): 2560-9.

Researcher : Chan YK

Project Title:	The signaling pathways of L-SIGN in response to ligand binding
Investigator(s):	Chan YK, Khoo US, Peiris JSM
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	03/2007

Abstract:

L-SIGN (liver/lymph node-specific ICAM-3 grabbing non-integrin) serves as a receptor for many of viral pathogens, including HIV1;2, HCV3, Ebola virus4, and SARS coronavirus (SARS-CoV)5. Its homologue, DC-SIGN, also serves as a receptor for many of the same viruses and has been utilized by some of the pathogens to escape immuno-surveillance. The binding sites of L-SIGN and DC-SIGN appear evolved to serve different functions. DC-SIGN mediates trafficking as a recycling receptor, releasing ligand at endosomal pH. In contrast, L-SIGN does not release ligands at endosomal pH nor does it mediate endocytosis, which indicates that it only functions as a binding receptor6. L-SIGN on transfected cells has been shown to internalize HCV virus-like particles and traffic to either lysosomal or non-lysosomal compartment depending on the cell type7. We speculate that L-SIGN may have other un-explored functions and signaling events after L-SIGN engagement with its ligand. Mitogen-activated protein kinases (MAPKs) are signal transducers that respond to extracellular stimulations, such as cytokines and viral infection. They in turn regulate cell differentiation, proliferation, survival and apoptosis8-11. There are three distinct MAPK cascades, extracellular signal-regulated kinases (ERK1/2), c-JUN N-terminal kinases (JNK), and p38/MAPK. Phosphatidylinositol 3-kinase (PI3K) signaling pathway also plays an important role in various cellular processes including cell growth and survival, vesicular trafficking, etc12. One of the key signaling molecules in the pathway is AKT which phosphorylates targets including GSK-3, FKHR-L1 and BAD. SARS-CoV infected permissive Vero E6 cells (which express the SARS-CoV receptor, ACE213) has been demonstrated to activate the MAPK and PI3K/Atk signaling pathways14-16. p38/MAPK has been shown to be activated during SARS-CoV viral replication14 in the infected cells, decreasing anti-apoptotic activity15. JNK and PI3K/Akt have been found to be important for the establishment of persistence in Vero E6 cells17. All these suggest that signaling pathways of MAPK and PI3K play important roles in regulating cellular responses to viral infection. To date, studies on DC-SIGN signaling are very limited while L-SIGN-mediated signaling has not been documented. Within the cytoplasmic domain of L-SIGN, it shares with DC-SIGN the potential internalization motifs, such as the di-leucine motif, etc.18. In DC-SIGN, the di-leucine motif is essential for receptor internalization19 and it is believed that L-SIGN also shares the same characteristics. A recent study has shown that DC-SIGN engagement on monocyte derived dendritic cells leads to phosphorylation of AKT and ERK20. DC-SIGN ligation has been shown to result in the activation of PI3K and MAPK signaling pathways20. It is possible that L-SIGN may also be involved in similar pathways; hence we will investigate if the presence of L-SIGN may modify MAPK and PI3K signaling activated by viral infection. The extra-cellular neck domain of L-SIGN is encoded by tandem repeats (from 3 to 9 repeats), 7 being predominant in the population. This neck region repeat is important for oligomerization of L-SIGN on the cell surface, which brings the carbohydrate recognition domain which it supports into proximity for high-affinity binding. We have shown that L-SIGN is a binding receptor for SARS-CoV and that heterozygotes in which the tandem-neck repeat lengths differ, have reduced binding for SARS-CoV when compared with homozygotes. Carriers with homozygous repeats were associated with a reduced risk for SARS infection.5 This is supported by our in-vitro observations that homozygous, but not heterozygous L-SIGN, possesses a protective role in reducing the final total viral titer in cultures with permissive cells. In part this may be attributed to a higher binding capacity of homozygous L-SIGN with greater cell association of virons, increased proteasome-dependent viral degradation and consequently a lower capacity for trans infection. This leads us to ask whether hetero- or homo-dimerization of L-SIGN binding with the ligands affects MAPK and PI3K signaling pathways. Objective 1: To investigate whether L-SIGN SARS-CoV binding affects MAPK and PI3K signaling pathways Objective 2: To investigate whether differences in hetero- and homo-dimerization of L-SIGN SARS-CoV binding affect and MAPK and PI3K signaling pathways Reference List 1. Pohlmann S, Soilleux EJ, Baribaud F et al. DC-SIGNR, a DC-SIGN homologue expressed in endothelial cells, binds to human and simian immunodeficiency viruses and activates infection in trans. Proc.Natl.Acad.Sci.U.S.A 2001;98:2670-2675. 2. Bashirova AA, Geijtenbeek TB, van Duijnhoven GC et al. A dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN)-related protein is highly expressed on human liver sinusoidal endothelial cells and promotes HIV-1 infection. J.Exp.Med. 2001;193:671-678. 3. Pohlmann S, Zhang J, Baribaud F et al. Hepatitis C virus glycoproteins interact with DC-SIGN and DC-SIGNR. J.Virol. 2003;77:4070-4080. 4. Alvarez CP, Lasala F, Carrillo J et al. C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans. J.Virol. 2002;76:6841-6844. 5. Chan VS, Chan KY, Chen Y et al. Homozygous L-SIGN (CLEC4M) plays a protective role in SARS coronavirus infection. Nat.Genet. 2006;38:38-46. 6. Guo Y, Feinberg H, Conroy E et al. Structural basis for distinct ligand-binding and targeting properties of the receptors DC-SIGN and DC-SIGNR. Nat.Struct.Mol.Biol. 2004;11:591-598. 7. Ludwig IS, Lekkerkerker AN, Depla E et al. Hepatitis C virus targets DC-SIGN and L-SIGN to escape lysosomal degradation. J.Virol. 2004;78:8322-8332. 8. Chang L, Karin M. Mammalian MAP kinase signalling cascades. Nature 2001;410:37-40. 9. Kyriakis JM, Avruch J. Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev. 2001;81:807-869. 10. Garrington TP, Johnson GL. Organization and regulation of mitogen-activated protein kinase signaling pathways. Curr.Opin.Cell Biol. 1999;11:211-218. 11. Whitmarsh AJ, Davis RJ. A central control for cell growth. Nature 2000;403:255-256. 12. Cantley LC. The phosphoinositide 3-kinase pathway. Science 2002;296:1655-1657. 13. Li W, Moore MJ, Vasilieva N et al. Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 2003;426:450-454. 14. Mizutani T, Fukushi S, Saijo M, Kurane I, Morikawa S. Phosphorylation of p38 MAPK and its downstream targets in SARS coronavirs-infected cells. Biochem.Biophys.Res.Commun. 2004;319:1228-1234. 15. Mizutani T, Fukushi S, Murakami M et al. Tyrosine dephosphorylation of STAT3 in SARS coronavirus-infected Vero E6 cells. FEBS Lett. 2004;577:187-192. 16. Mizutani T, Fukushi S, Saijo M, Kurane I, Morikawa S. Importance of Akt signaling pathway for apoptosis in SARS-CoV-infected Vero E6 cells. Virology 2004;327:169-174. 17. Mizutani T, Fukushi S, Saijo M, Kurane I, Morikawa S. JNK and PI3k/Akt signaling pathways are required for establishing persistent SARS-CoV infection in Vero E6 cells. Biochim.Biophys.Acta 2005;1741:4-10. 18. Koppel EA, van Gisbergen KP, Geijtenbeek TB, van KY. Distinct functions of DC-SIGN and its homologues L-SIGN (DC-SIGNR) and mSIGNR1 in pathogen recognition and immune regulation. Cell Microbiol. 2005;7:157-165. 19. Sol-Foulon N, Moris A, Nobile C et al. HIV-1 Nef-induced upregulation of DC-SIGN in dendritic cells promotes lymphocyte clustering and viral spread. Immunity. 2002;16:145-155. 20. Caparros E, Munoz P, Sierra-Filardi E et al. DC-SIGN ligation on dendritic cells results in ERK and PI3K activation and modulates cytokine production. Blood 2006;107:3950-3958.

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Chan Y.K., Contribution of Genetic Variants to Disease Risk, Department of Pathology, The University of Hong Kong. 2008.

Chan Y.K., Genetic Association Studies: Applications in Cancer Research, 14th Hong Kong International Cancer Congress. 2007.

Chen Y., Chan V.S.F., Zheng B., Chan Y.K., Xu X., To Y.F., Huang F.P., Khoo U.S. and Lin C.L., A novel subset of putative stem/progenitor CD34+Oct-4+ cells is the major target for SARS coronavirus in human lung., J Exp Med.. 2007, 204: 2529-36.

Khoo U.S., Chan Y.K., Chan V.S. and Lin C.L., DC-SIGN and L-SIGN: the SIGNs for infection, J Mol Med. 2008.

Khoo U.S., Chan Y.K., Chan V.S.F., Ching C.Y.J., Yam L., Chu C.M., Lai S.T., Wong T.Y., Tam P.K.H., Yip S.P., Leung G.M., Lin C.L. and Peiris J.S.M., Role of polymorphisms of the inflammatory response genes and DC-SIGNR in genetic susceptibility to SARS and other infections, Hong Kong Med J. 2008, 14(4)(Suppl 4): S9-12.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Liu S., Tsang P.C.K., Chan Y.K., Cheung A.N.Y., Chan K.K.L., Leung C.Y. and Ngan H.Y.S., Distribution of Six Oncogenic Types of Human Papillomavirus and Type 16 Integration Analysis in Chinese Women with Cervical Precancerous Lesions and Carcinomas, Tumour Biol. 2008, 29(2): 105-113.

Ngan H.Y.S., Tsang P.C.K., Chan Y.K., Liu S., Cheung A.N.Y., Chan K.K.L. and Leung C.Y., Human papillomavirus genotyping and integration in cervical cancers and precursor lesions, 24th International Papillomavirus Conference and Clinical Workshop. Beijing, China, November. 2007.

Ngan H.Y.S., Liu S., Leung T.W., Cheung A.N.Y. and Chan Y.K., Study of human papillomavirus status in Southern Chinese women with normal cervix, precancerous cervical lesions and cervical cancers. , Health Research Symposium 2007: Building Bridges between research, practice and policy. September, Hong Kong. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Li A.S.M., Chan Y.K. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress 2007, 14-16 Nov. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Wong K.Y., Li A.S.M., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Oct4 is Epigenetically Regulated by Methylation in Normal Placenta and Gestational Trophoblastic Disease, Placenta. 2008, 29(6): 549-554.

Researcher : Chan YP

List of Research Outputs

Yuen H.F., Chan Y.P., Chan K.K., Chu Y.Y., Wong M.L.Y., Law S.Y.K., Srivastava G., Wong Y.C., Wang X. and Chan K.W., Id-1 and Id-2 are markers for metastasis and prognosis in oesophageal squamous cell carcinoma, Br J Cancer. 2007, 97(10): 1409-15.

Researcher : Chan YW

List of Research Outputs

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Suehiro Y., Wong C.W., Chirieac L.R., Kondo Y., Shen L., Webb C.R., Chan Y.W., Chan A.S.Y., Chan T.L., Wu T.T., Rashid A., Hamanaka Y., Hinoda Y., Shannon R.L., Morris J., Issa J.P., Yuen S.T., Leung S.Y. and Hamilton S.R., Epigenetic-Genetic Interactions in the APC/WNT, RAS/RAF, and P53 Pathways in Colorectal Carcinoma, Clin Cancer Res. 2008, 14(9): 2560-9.

Researcher : Chen WYW

List of Research Outputs

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Ying J., Li H., Chen W.Y.W., Srivastava G., Gao Z. and Tao Q., WNT5A is epigenetically silenced in hematologic malignancies and inhibits leukemia cell growth as a tumor suppressor, Blood. 2007, 110(12): 4130-1.

Researcher : Chen Y

List of Research Outputs

Chen Y.X., Man K., Ling G.S., Chen Y., Sun B., Cheng Q., Wong O.H., Lo C.K., Ng I.O.L., Chan L.C., Lau G., Lin S.C.L., Huang F. and Huang F.P., A crucial role for dendritic cell (DC) IL-10 in inhibiting successful DC-based immunotherapy: superior antitumor immunity against hepatocellular carcinoma evoked by DC devoid of IL-10, Journal of Immunology. 2007, 179(9): 6009-6015.

Researcher : Cheung ANY

Project Title:	Stem cell related genes in gestational trophoblastic diseases
Investigator(s):	Cheung ANY, Ngan HYS, Chan YK, Yeung WSB
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2005

Abstract:

To characterize the expression levels and epigenetic alterations of important stem-cell related genes in gestational trophoblastic disease (GTD) in an attempt to evaluate the roles of such genes in the pathogenesis of GTD; and to define useful genetic markers as predictors of clinical progression or targets for therapy.

Project Title:	Selecting most suitable students for production of best medical doctors-which criteria should we use?
Investigator(s):	Cheung ANY, Patil NG, Ip MSM, Chan LC
Department:	Pathology
Source(s) of Funding:	Leung Kau Kui Research and Teaching Endowment Fund - Teaching Grants
Start Date:	02/2005

Abstract:

(a) enhancing the processes for evaluating and improving admission criteria for MBBS curriculum; and potentially; (b) improving curriculum design and (c) improving learning opportunities of students on ethical values and communication skills.

Project Title:	RAS association domain family related genes in gynaecological cancers
Investigator(s):	Cheung ANY, Ngan HYS, Liu VWS, Siu KY, Ip PPC
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	01/2006

Abstract:

The purpose of the proposed investigation are to characterize the expression levels and epigenetic alterations of RAS association domain family related genes in ovarian and endometrial cancers in an attempt to evaluate the roles of such genes in the their carcinogenesis; and to define useful genetic markers as predictors of clinical progression or targets for therapy. Key Issues and Problems being addressed: Endometrial and ovarian cancers are two common cancers in the female genital tract and the latter is responsible for the highest mortality in this group of cancers. Early detection is essential since the prognosis is significantly affected by the staging of the cancers at first diagnosis. We have earlier reported the significant roles of a few tumour suppressor genes in gynaecological malignancies (1-3) as an attempt to explore their application as potential cancer markers and to better our understanding on endometrial and ovarian cancers carcinogenesis. RASSF1 (the RAS association domain family 1) and RASSF2 are members of a new group of RAS effectors which may function as tumor suppressors genes and interact with the K-Ras oncogene affecting apoptosis and cell cycle arrest (4). Whilst mutation of RASSF1A (isoform A) occurs rarely in human cancers, RASSF1A methylation has been reported in at least 37 tumor types and is postulated to have the potential to be used as a marker for tumor diagnosis, early detection and monitoring (5). RASSF2A (isoform A) was recently reported to be frequently inactivated in colorectal cancer by CpG island promoter hypermethylation (6). The human DOC-2/DAB2 interactive protein gene (hDAB2IP) is a novel member of the Ras GTPase-activating gene family (7, 8). hDAB2IP directly interacts with the N-terminal domain of disabled-2 protein (DAB2, also known as DOC-2, differentially expressed in ovarian carcinoma 2), which appears to be a tumor suppressor in various cancers (9). Both DAB2IP and DAB2 form a unique protein complex that has a negative regulatory effect on the Ras-mediated signal pathway (8). hDAB2IP methylation was found to be associated with cancers of the prostate, breast, lung and gastrointestinal tract (10, 11). BLU gene is a candidate tumor suppressor gene immediately upstream of RASSF1 (12). Heterogenous association between methylation and expression status of BLU gene in various cancers was observed. The roles of these RAS association domain family related genes in endometrial and ovarian cancers are still unknown. References (*Publications from our group): *1. Chan KY, Ozcelik H, Cheung AN, Ngan HY, Khoo US. Epigenetic factors controlling the BRCA1 and BRCA2 genes in sporadic ovarian cancer. Cancer Res. 2002 Jul 15;62(14):4151-6. *2. Chan QK, Khoo US, Chan KY, Ngan HY, Li SS, Chiu PM, Man LS, Ip PP, Xue WC, Cheung AN. Promoter methylation and differential expression of pi-class glutathione S-transferase in endometrial carcinoma. J Mol Diagn. 2005;7:8-16. *3. Shen DH, Chan KY, Khoo US, Ngan HY, Xue WC, Chiu PM, Ip P, Cheung AN. Epigenetic and genetic alterations of p33ING1b in ovarian cancer. Carcinogenesis. 2005;26:855-63. 4. Vos MD, et al. RAS uses the novel tumor suppressor RASSF1 as an effector to mediate apoptosis. J Biol Chem 2000; 275: 35669-35672. 5. Agathanggelou A, et al. Methylation associated inactive of RASSF1A from region 3p21.3 in lung, breast and ovarian tumors. Oncogene, 2001, 20:1509-1518. 6. Hesson LB, et al. CpG island promoter hypermethylation of a novel Ras-effector gene RASSF2A is an early event in colon carcinogenesis and correlates inversely with K-ras mutations. Oncogene, 2005, 24:3987-3994. 7. Chen H, et al. Differential regulation of human gene DAB2IP in normal and malignant prostatic epithelia: cloning and characterization. Genomics 2002; 79: 573-581. 8. Wang Z, et al. The mechanism of growth-inhibitory effect of DOC-2/DAB2 in prostate cancer. Characterization of a novel GTPase-activating protein associated with N-terminal domain of DOC-2/DAB2. J Biol Chem 2002; 277: 12622-12631. 9. Zhou J, Hsieh JT. The inhibitory role of DOC-2/DAB2 in growth factor receptor-mediated signal cascade. DOC-2/DAB2-mediated inhibition of ERK phosphorylation via binding to Grb2. J Biol Chem 2001; 276: 27793-27798. 10. Dote H, et al. Aberrant promoter methylation in human BAB2 interactive protein (hDAB2IP) gene in breast cancer. Clin Cancer Res, 2004, 10: 2082-2089. 11. Yano M, et al. Aberrant promoter methylation of human DAB2 interactive protein (hDAB2IP) gene in lung cancers. Int J Cancer, 2005, 113: 59-66. 12. Agathanggelou A, et al. Epigenetic inactivation of the candidate 3p21.3 suppressor gene BLU in human cancers. Oncogene. 2003, 22(10):1580-8.

Project Title:	Folate and folate receptor alpha in ovarian cancers
Investigator(s):	Cheung ANY, Siu KY
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	01/2007

Abstract:

The project aims at investigating the role of folate and folate receptor alpha in affecting the growth and invasive behaviors of ovarian cancers. World wide, ovarian cancer is a common cancer in women, contributing to the highest mortality among all gynecological cancers. More importantly, there is a recent significant increase in the incidence, rising from 9.8 per 100,000 (crude incidence rate) in 1999 to 12.5 in 2002 [1]. Recent studies in our laboratories have reported several tumor suppressor genes as potential cancer markers, which play a vital role in gynecological malignancies [2-6]. Several epidemiologic factors for this gynecological cancer have been documented. Age, a family history of ovarian or breast cancer, nulliparity and use of oral contraceptive are associated with the risk of ovarian cancer [7]. Recently, attention has focused on the association between high intakes of micronutrients such as folate, vitamin C, vitamin E and lower risk of various cancers [8]. For instance, a high dietary folate intake is related to lower risk of ovarian cancer, particularly among women who consume alcohol [9-11]. Folate, a water-soluble B vitamin found in most vegetable, is required for DNA synthesis, and the related methionine metabolic pathway is essential for DNA methylation. Folate deficiency has found to induce DNA hypomethylation, alter gene expression and misincorporate uracil into DNA leading to chromosome damage, all of which are key factors for carcinogenesis [12,13]. Pregnancy, lactation and alcohol abuse are some common causes for the increase of folate requirements in the body. It is particularly important to notice that alcohol consumption is considered as a major factor account for the inverse relation between folate intake and ovarian cancer risk [10,14,15]. It is interested to note that the folate receptor alpha (FR alpha), a single chain glycosyl-phosphatidylinositol–anchored membrane protein, was found to be overexpressed in nonmucinous ovarian tumors and its expression was directly associated with the progression of ovarian cancer, implicating that uptake of folate by tumor cells may confer tumor growth [16-18]. Such observation suggests a hypothesis that folate may involve in the growth of both normal and tumor cells. Furthermore, the FR alpha was shown to be up-regulated when cells are exposed to folate-deplete media and down-regulated in folate-replete media [19-21]. While the expression of the FR alpha in ovarian cancer among Oriental, such as in Hong Kong, is still unknown, it is worthy to include local data to investigate the hypothesis stated. REFERENCES 1. Hong Kong Department of Health. (2002) The Hong Kong Cancer Registry: Annual Report 2001. *2. Chan, K.Y.K., et al. (2002) Epigenetic factors controlling the BRCA1 and BRCA2 genes in sporadic ovarian cancer. Cancer Research, 63, 4151-4156. *3. Chan, Q.K.Y., et al. (2005) Promoter Methylation and Differential Expression of pi-class Glutathione-S-Transferase in Endometrial Carcinoma. Journal of Molecular Diagnostics, 7, 8-16. *4. Chan, Q.K.Y., et al. (2005) Single Nucleotide Polymorphism of Pi-class Glutathione-S-Transferase and Susceptibility to Endometrial Carcinoma. Clin. Cancer Res., 11, 2981-2985. *5. Shen, D.H., et al. (2005) Epigenetic and genetic alterations of p33ING1b in ovarian cancer. Carcinogenesis, 26, 855-863. *6. Yang, C.Q., et al. (2006) Single nucleotide polymorphisms of follicle-stimulating hormone receptor are associated with ovarian cancer susceptibility. Carcinogenesis, 27, 1502-6. 7. Lukanova, A. and Kaaks, R. (2005) Endogenous hormones and ovarian cancer: epidemiology and current hypotheses. Cancer Epidemiol. Biomarkers Prev., 14, 98-107. 8. Woodside, J.V., et al. (2005) Micronutrients: dietary intake v. supplement use. Proc. Nutr. Soc., 64, 543-553. 9. Larsson, S.C., et al. (2004) Dietary Folate Intake and Incidence of Ovarian Cancer: The Swedish Mammography Cohort. J. Natl. Cancer Inst., 96, 396-402. 10. Kelemen, L.E., et al. (2004) Association of folate and alcohol with risk of ovarian cancer in a prospective study of postmenopausal women. Cancer Causes Control, 15, 1085–1093. 11. Larsson, S.C. and Wolk, A. (2004) Wine consumption and epithelial ovarian cancer. Cancer Epidemiol. Biomarkers Prev., 13, 1823–1824. 12. Blount, B.C., et al. (1997) Folate deficiency causes uracil misincorporation into human DNA and chromosome breakage: implications for cancer and neuronal damage. Proc. Natl. Acad. Sci., 94, 3290-3295. 13. Duthie, S.J. (1999) Folic acid deficiency and cancer: mechanisms of DNA instability. Br. Med. Bull., 55, 578-592. 14. Kelemen, L.E., et al. (2005) Multivitamin and alcohol intake and folate receptor alpha expression in ovarian cancer. Cancer Epidemiol. Biomarkers Prev., 14, 2168-2172. 15. Pelucchi, C., et al. (2005) Dietary folate, alcohol consumption, and risk of ovarian cancer in an Italian case-control study. Cancer Epidemiol. Biomarkers Prev., 14, 2056-2058. 16. Ross, J.F., et al. (1994) Differential regulation of folate receptor isoforms in normal and malignant tissues in vivo and in established cell lines. Physiologic and clinical implications. Cancer, 73, 2432–2443. 17. Campbell, I.G., et al. (1991) Folate-binding protein is a marker for ovarian cancer. Cancer Res., 51, 5329–5338. 18. Toffoli, G., et al. (1997) Overexpression of folate binding protein in ovarian cancers. Int. J. Cancer, 74, 193–198. 19. Luhrs, C.A., et al. (1992) Transfection of a glycosylated phosphatidylinositol-anchored folate-binding protein complementary DNA provides cells with the ability to survive in low folate medium. J. Clin. Invest., 90, 840-847. 20. Matsue, H., et al. (1992) Folate receptor allows cells to grow in low concentrations of 5-methyltetrahydrofolate. Proc. Natl. Acad. Sci. U. S. A., 89, 6006–6009. 21. Hsueh, C.T. and Dolnick, B.J. (1993) Altered folate binding protein mRNA stability in KB cells grown in folate-deficient medium. Biochem. Pharmacol., 45, 2537–2545.

Project Title:	Akt and p21-activated kinase signaling pathways in gestational trophoblastic disease
Investigator(s):	Cheung ANY, Ngan HYS, Siu KY
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2007

Abstract:

To elucidate the roles of Akt and p21-activated serine/threnonine kinases (Paks) and their related genes in the pathogenesis of gestational trophoblastic diseases; to evaluate the potential of these genes as markers for predicting clinical progression and molecular targets for therapy.

Project Title:	Cervical cancer screening by enhanced cervical cytology-application of novel markers
Investigator(s):	Cheung ANY, Ngan HYS, Guan XY
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	03/2007

Abstract:

To explore with application of novel markers, some generated by our local genetic studies, to be detected by immunohistochemistry and ISH as an adjunct to enhance sensitivity and specificity of liquid based cervical cytology.

List of Research Outputs

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Chan K.K.L., Wei N., Liu S., Liao X., Cheung A.N.Y. and Ngan H.Y.S., Estrogen receptor subtypes in ovarian cancer: a clinical correlation, Obstet Gynecol. 2008, 111: 144-151.

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Cheung A.N.Y., Tsun O.K.L., Szeto E.F., Wong G., Lo S. and Ngan H.Y.S., Application of Novel Markers to Cervial Cytology for enhancing Cervical Cancer Screening, Annual Scientific Meeting of Hong Kong Society of Cytology, Hong Kong, 8-9 December. 2007.

Cheung A.N.Y., Human embryonic stem cell genes in gestational trophoblastic diseases, XIX World Congress On Gestational Trophoblastic Diseases, Fukuoka, 11-14 November. 2007.

Cheung A.N.Y., Tsun O.K.L. and Szeto E.F., ThinPrep Imaging System in Cervical Cytology Screening, Annual Scientific Meeting of Hong Kong Society of Cytology, Hong Kong, 8-9 December. 2007.

Cheung A.N.Y., p21-activated kinase (PAK) in gestational trophoblastic neoplasia, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Over-expression of Prostatic Stem Cell Antigen (PSCA) is associated with Gestational Trophoblastic neoplasia. , Histopathology. 2007, 52: 167-174.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Overexpression of prostate stem cell antigen is associated with gestational trophoblastic neoplasia, Histopathology. 2008, 52(2): 167-174.

Ip P.P.C., Lam K.W., Cheung C.L., Yeung C.W., Pun T.C., Chan K.Y.Q. and Cheung A.N.Y., Tranexamic Acid-associated Necrosis and Intralesional Thrombosis of Uterine Leiomyomas: A Clinicopathologic Study of 147 Cases Emphasizing the Importance of Drug-induced Necrosis and Early Infarcts in Leiomyomas , American Journal of Surgical Pathology. 2007, 31(8): 1215-1224.

Kwan T.T.C., Chan K.K.L., Yip M.W., Tam K.F., Cheung A.N.Y., Young P.M.C., Lee P.W.H. and Ngan H.Y.S., Barriers and facilitators to human papillomavirus vaccination among Chinese adolescent girls in Hong Kong: a qualitative–quantitative study, Sex Transm Infect. 2008, 84(3): 227-232.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Liao X., Siu K.Y., Au W.H., Wong E.S.Y., Ngan H.Y.S. and Cheung A.N.Y., Aberrant Activation of Hedgehog Signaling Pathway in Ovarian Cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Liu S., Tsang P.C.K., Chan Y.K., Cheung A.N.Y., Chan K.K.L., Leung C.Y. and Ngan H.Y.S., Distribution of Six Oncogenic Types of Human Papillomavirus and Type 16 Integration Analysis in Chinese Women with Cervical Precancerous Lesions and Carcinomas, Tumour Biol. 2008, 29(2): 105-113.

Ngan H.Y.S., Tsang P.C.K., Chan Y.K., Liu S., Cheung A.N.Y., Chan K.K.L. and Leung C.Y., Human papillomavirus genotyping and integration in cervical cancers and precursor lesions, 24th International Papillomavirus Conference and Clinical Workshop. Beijing, China, November. 2007.

Ngan H.Y.S., Kwan T.T.C., Tam K.F., Chan K.K.L., Young P.M., Lo S.S., Cheung A.N.Y. and Lee P.W.H., Knowledge and attitute of Chinese women on cervical cancer and human papillomavirus, AOCOG 2007 Golden Jubilee of Federation, The XXth Asian and Oceanic Congress of Obstetrics and Gynaecology, September 21-25, 2007, Tokyo, Japan. 2007.

Ngan H.Y.S., Liu S., Leung T.W., Cheung A.N.Y. and Chan Y.K., Study of human papillomavirus status in Southern Chinese women with normal cervix, precancerous cervical lesions and cervical cancers. , Health Research Symposium 2007: Building Bridges between research, practice and policy. September, Hong Kong. 2007.

Pon Y.L., Zhou H., Cheung A.N.Y., Ngan H.Y.S. and Wong A.S.T., p70 S6 kinase promotes epithelial to mesenchymal transition through Snail induction in ovarian cancer cells, Cancer Research. 2008, 68(16): 6524-6532.

Siu K.Y., Chan H.Y., Kong S.H., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Differential expression of folate receptor alpha and reduced folate carrier and effect of folate in ovarian cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Siu K.Y., Woo N.W., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th Annual Growth Factor and Signal Transduction Symposium titled "Senescence, Aging and Cancer",Iowa, 26-29 July . 2007.

Woo N.W.S., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th annual Growth Factor and Signal Transduction Symposium, Iowa, 26-29 July. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Li A.S.M., Chan Y.K. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress 2007, 14-16 Nov. 2007.

Zhang H., Siu K.Y., Li A.S.M., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Wong K.Y., Li A.S.M., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Oct4 is Epigenetically Regulated by Methylation in Normal Placenta and Gestational Trophoblastic Disease, Placenta. 2008, 29(6): 549-554.

Researcher : Chiang AKS

Project Title:	Experimental immunotherapy of tumours
Investigator(s):	Chiang AKS
Department:	Paediatrics & Adolescent Med
Source(s) of Funding:	Research Initiation Programme
Start Date:	04/2001

Abstract:

To develop strategies in enhancing the host immune response against tumours.

Project Title:	Genetic studies of tubercubosis
Investigator(s):	Chiang AKS
Department:	Paediatrics & Adolescent Med
Source(s) of Funding:	Other Funding Scheme
Start Date:	03/2002

Abstract:

To host susceptibility genes in tubercubosis; to study pharmacogenomics in anti-tubercubosis drug metabolism.

Project Title:	Prospective study of virologic and immunologic parameters of primary Epstein-Barr virus infection in Chinese children
Investigator(s):	Chiang AKS, Chan KH
Department:	Paediatrics & Adolescent Med
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	10/2003

Abstract:

An unresolved issue in EBV immunology concerns the relationship between clinical IM-like symtoms, virus DNA load and CD8+ T cell lymphocytosis; Acute IM in adolescents is associated with large expansions of EBV lytic epitope reactivities which are followed by a later expansion of latent epitope responses. The magnitude of the EBV lytic and latent epitope reactivities would be studied and compared in both IM and non-IM patients; likewise, is there a difference in the rate of emergence of latent epitope responses bewteen the IM and non -IM patients? The kinetics and evolution of the EBV epitope-specific cytotoxic T-lymphocyte (CTL) responses from primary to persistent phases of EBV infection would be compared between the two groups of children.

Project Title:	Prospective study of Epstein-Barr virus (EBV) strains in primary EBV infection
Investigator(s):	Chiang AKS
Department:	Paediatrics & Adolescent Med
Source(s) of Funding:	Small Project Funding
Start Date:	11/2004

Abstract:

To study the occurrence of Epstein-Barr virus (EBV) diversity and coinfection in a prospective study of primary Epstein-Barr virus infection in Chinese children.

Project Title:	Longitudinal study of Epstein-Barr virus specific antibody responses in childhood infectious mononucleosis
Investigator(s):	Chiang AKS
Department:	Paediatrics & Adolescent Med
Source(s) of Funding:	Small Project Funding
Start Date:	12/2005

Abstract:

The purpose of the proposed study is to compare the evolution of Epstein-Barr virus (EBV) specific antibody responses in a longitudinal followup study of symptomatic and asymptomatic primary EBV infection. What is known about EBV antibody responses in the literature? Epstein-Barr virus (EBV) infects greater than 90% of all human populations and establishes a lifelong persistence within the host. Primary infections by EBV may remain silent or induce a self-limiting lymphoproliferative disorder, infectious mononucleosis (IM). EBV-encoded antigens are divided into those expressed in the viral replicative cycle, i.e., the viral capsid antigen (VCA) and the early amtigens (EA) of the diffuse (D) and the restricted (R) varieties and those expressed in the viral latent cycle, the EBV-determined nuclear antigen (EBNA). The different timing of emergence between the antibodies to VCA and EA and those to EBNA indicates that the two sets of antigens are likely expressed on separate types of cells which become available to stimulate antibody responses under different circumstances. The laboratory diagnosis of EBV infection is based on serological tests to detect both specific antibodies to EBV antigens (1, 2) and heterophile antibodies (3, 4). At the onset of symptoms of IM, the humoral response to EBV infection is characterized by the presence of VCA IgM antibodies and the concurrent rise of VCA IgG antibodies (5). Antibodies to diffuse early antigen (anti-EA-D) are transitory and are detected in about 80% of infected persons. Antibodies to EA-R may become detectable in protracted cases of the disease after the anti-D titres have subsided. Anti-EBNA IgG can usually be detected at a later stage at several weeks to months after the the onset of symptoms. The majority of adult IM patients also develop IgM heterophile antibodies of the Paul-Bunnell type but the reason for their transient emergence is unknown. However, these heterophile antibodies are absent in the majority of children under 4 years of age (6). The antibody responses in silent primary EBV infections (more frequently occurred in children than adults) largely conform to those seen in IM except that antibodies to the EA complex are directed against R instead of D (7, 8). The early EBV-specific IgG antibody response after primary infection is made up of low avidity antibodies. The avidity increases in the few weeks to several months after the infection (9, 10). This property of the humoral IgG response has been employed in serological assays to distinguish between primary and re-infection or reactivation of different viruses such as rubella, CMV and parvovirus (11, 12, 13). Key issues: The pattern of antibody responses and the antibody levels to EBV-encoded antigens are largely derived from cross sectional studies of large number of individuals. Longitudinal serology data of persons recovering from IM is seldom available and information on the humoral response in asymptomatic EBV infection is scanty. I have followed a large number of Chinese children presenting with infectious mononucleosis and some children who are found to have asymptomatic primary EBV infection from the time of onset of infection through convalescence to persistence. It offers an opportunity to compare the natural evolution of antibody responses against EBV in symptomatic and asymptomatic infections. Project objectives: 1. To define the evolution of the humoral response against a human persistent virus. 2. To study the maturation of the humoral response against a persistent virus in terms of antibody avidity. 3. To compare the magnitude and the quality of the humoral response against viral lytic and latent EBV antigens from acute phase through convalescent to persistent phase in symptomatic and asymptomatic infections. References: 1.Henle W and Henle G: Epstein-Barr virus specific serology in immunologically compromized individuals. Cancer Res 41: 4222-4225, 1981 2.Henle W and Henle G: Immunology of Epstein-Barr virus. In: B, Roizman (Ed.), The Herpsesviruses, Vol. 1. Plenum Publishing Corp., Ne York, pp 209-252, 1982 3.Paul JR and Bunnell WW: The presence of heterophile antibodies in infecetious mononucleosis. Am J Med Sci 183: 90-104, 1932 4.Davidsohn I and Lee CL: The clinical serology of infectious mononucleosis. In: RL Carter and HG Penman (Eds.), Infectious Mononucleosis. Blackwell Scientific Publications, Ltd., Oxford, pp 177-200, 1969 5.Okano M, Thiele GM, Davis JR, Grierson HL, Purtilo DT: Epstein-Barr virus and human diseases: recent advances in disgnosis. Clin Microbiol Rev 1: 300-312, 1988 6.Sumaya CV: Infectious mononucleosis and other EBV infections: diagnostic factors. Lab Manag 24: 37-45, 1986 7.Biggar RJ, Henle G, Bocker J, Lennette ET, Fleisher G, Henle W: Primary Epstein-Barr virus infections in African infacnts. II. Clinical and serological obsevations during seroconversion. Int J cancer 22: 244-250, 1978 8.Fleisher G, Henle W, Henle G, Lennette ET, Biggar RJ: Primary Epstein-Barr virus infection in American infants: clinical and serological observations. J Infect Dis 139: 553-558, 1979 9.Inouye S, Hasegawa A, MAtsuno S, Katwo S: Changes in antibody avidity after virus infections: detection by an immunosorbent assay in which a mild protein-denaturing agent is employed. J Clin Microbiol 20: 525-529, 1984 10.Kocks C and Rajewsky K: Stepwise intracloncal maturation of antibody affinity through somatic hypermutation. Proc Nat Acad Sci USA 85: 8206-8210, 1988 11.Thomas HIJ and Morgan-Capner P: Rubella-specific IgG subclass avidity and its role in differenetiation between primary rubella and rubella reinfection. Epidemiol Infect 101: 591-598, 1988 12.Lutz E, Ward KN, Gray JJ: Maturation of antibody avidity after primary human cytomegalovirus infection is delayed in immunosuppressed solid organ transplant patients. J Med Virol 44: 317-322, 1994 13.Gray JJ, Cohen BJ, Dresselberger U: Detection of human parvirus B19-specific IgM and IgG sntibodies using a recombinant viral VP1 antigen expressed in insect cells and estimation of time of infection by testing for antibody avidity. J Virol Methods 44: 11-24, 1993

Researcher : Ching CYJ

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Khoo U.S., Chan Y.K., Chan V.S.F., Ching C.Y.J., Yam L., Chu C.M., Lai S.T., Wong T.Y., Tam P.K.H., Yip S.P., Leung G.M., Lin C.L. and Peiris J.S.M., Role of polymorphisms of the inflammatory response genes and DC-SIGNR in genetic susceptibility to SARS and other infections, Hong Kong Med J. 2008, 14(4)(Suppl 4): S9-12.

Researcher : Ching YP

Project Title:	Roles and regulation of group II p21-activated protein kinases:-implications in cancer metastasis
Investigator(s):	Ching YP, Jin D, Ng IOL
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2005

Abstract:

To study: (1) Characterisation of the interaction between Pak5 and NM23 i) co-immunoprecipitation of Pak5 adn NM23 ii) defining the binding domain between Pak 5 and NM23 iii) xploring the interaction between Pak4 adn NM23. (2) Impact of Pak5-NM23 interaction in the biochemical properties of Pak5 and NM23 i) nucleotide diphosphate kinase activity ii) GTPase activating activity iii) in vitro kinase activity iv) subcellular localisation. 3) Roles of PakII in cancer metastasis using HCC as a model i) expression profile of Pak4 in HCC ii) clinicopathological analysis iii) cell invasion assay.

Project Title:	Roles of p21-activated protein kinase (Pak) 1 in the pathogenesis of liver cancer
Investigator(s):	Ching YP
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	07/2005

Abstract:

To characterize the overexpression of Pak1 protein and its activities in human HCC; to delineate the signaling pathways mediated by Pak1 in HCC; to investigate the roles of Pak1 in the metastasis of HCC.

Project Title:	Roles of p21-activated protein kinase (Pak) 1 in the pathogenesis of liver cancer
Investigator(s):	Ching YP, Ng IOL, Jin D, Yau TO
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2006

Abstract:

(1) To characterize the mechanisms leading to Pak1 overexpression in human HCC; (2) to delineate the roles of Pak1 in hepatocarcinogenesis: (i) Phosphorylation of possible downstream targets of Pak1 in human HCCs. (ii) characterization of the tumorigenic activity of Pak1 in HCC cells. (iii) characterization of the anti-apoptotic activity of Pak1 in HCC cells. (3) to investigate the role of Pak1 in cancer metastasis: >(i) regulation of cell motility and cell adhesion by Pak1 in HCC cells. (ii) HGF/Rac1/Cdc42/Pak1 signaling in HCC metastasis. (iii) HGF/Pak1 mediated angiogenic activity. (iii) HGF/Pak1 mediated angiogenic activity.</I

Project Title:	Functional characterization of a putative tumour suppressor, AMP-activated protein kinase, in liver cancer
Investigator(s):	Ching YP
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	02/2006

Abstract:

Purpose of study Liver cancer (hepatocellular carcinoma, HCC) is one of the most common cancers in the world, especially in Asia and Africa, and is the third most common fatal cancer in Hong Kong. While the risk factors are well defined, the underlying molecular mechanisms of HCC are still far from clear. Since the development of HCC is a multistep process, our long-standing interest is to identify tumour suppressor genes that play important roles in hepatocarcinogenesis. In our search, we have found that a hepatic kinase called AMP-activated protein kinase (AMPK), a key regulator for lipid and glucose metabolism in response to energy stress, is frequently downregulated in HCC cell lines and clinical samples (42%). Interestingly, recent discoveries have shown that tumour suppressors LKB1 and TSC2 lie upstream and downstream of AMPK, respectively, indicating that AMPK may be important in the regulation of cell growth, proliferation and apoptosis. Our pilot studies have also demonstrated that ectopic expression of the AMPK catalytic subunit in HepG2 cells significantly suppresses cell growth in colony formation assay. Conversely, HCC cell line with stable knockdown of AMPK catalystic subunit expression displays a much higher proliferation rate than that of the parental cell line (see research plan). These results suggest that AMPK possesses an activity to inhibit HCC cell growth. Here we propose to fully document the expression of AMPK and its effectors in HCC. We will explore the molecular basis of the underexpression of AMPK catalytic subunit in human HCC. We will also confirm the tumour suppressor activity of AMPK and delineate the molecular mechanisms by which loss of AMPK contributes to the formation of HCC. Findings derived from our work should shed important light on the pathogenesis of HCC and may provide novel targets for therapeutic intervention. Key issue and problem being address So far, the overall prognosis of HCC is unsatisfactory due to high incidence of recurrence and metastasis. It is therefore a high priority to unravel the molecular mechanisms underlying the pathogenesis of HCC so that better treatment modalities can be designed. In previous reports and our preliminary study, activation of AMPK has been demonstrated to play a role in suppressing the growth of cancer cells. These results prompt us to further address the pathogenic role of AMPK in HCC and to determine how dysregulation of AMPK leads to hepatocarcinogenesis. In this study, we will define the tumour suppressive function of AMPK in HCC and the regulatory roles of AMPK in cell signalling. Since AMPK is an important physiological regulator of cellular metabolism in response to nutrient stress and energy supply, our proposed study should advance our understanding on whether perturbation of energy metabolism can possibly linked to carcinogenesis. Results obtained in this study will derive novel insight on how the loss of AMPK function leads to the formation of cancer, aiming to provide opportunity for new molecular drug targets.

Project Title:	Molecular neurobiology: Regulation of p21-activated protein kinase 5 in neurodegenerative disease
Investigator(s):	Ching YP
Department:	Anatomy
Source(s) of Funding:	Matching Fund for NSFC Young Researcher Award
Start Date:	01/2007
Completion Date:	12/2009

Abstract:

To study molecular neurobiology: regulation of p21-activated protein kinase 5 in neurodegenerative disease.

Project Title:	Functional characterisation of a putative tumor suppressor gene, TAX1BP2, in liver cancer
Investigator(s):	Ching YP
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	04/2007

Abstract:

1. Key issues and problem being addressed: Liver cancer (hepatocellular carcinoma, HCC) is one of the most common cancers worldwide. The prognosis of HCC patients is often poor because of the delay in diagnosis and its high recurrence rate after surgery. Although the risk factors of HCC, such as hepatitis B and C virus infection, cirrhosis, and dietary aflatoxin are well established, the genetic mechanisms in the pathogenesis and tumour progression are poorly defined. Chromosome instability that leads to clonal expansion of genetically altered cells is a hallmark of cancer and is highly relevant to hepatocarcinogenesis. Thus characterization of tumor suppressor genes and elucidation of the mechanisms of chromosome instability are both major challenges in liver cancer research. 2. Purpose of proposed project: Recently, we have identified and characterized a novel cellular centrosomal protein, which we have named TAX1BP2 (Ching et al., 2006, Nature cell Biology 8: 717-24). We have demonstrated that TAX1BP2 plays an important role in the regulation of centrosome duplication, and dysregulation of TAX1BP2 may lead to aneuplody of cells. In our preliminary study, we observed that TAX1BP2 is frequently underexpressed in human HCC (40%) and the underexpression of TAX1BP2 transcript is significantly associated with a poorer prognosis in terms of shorter overall survival rates. In addition, TAX1BP2 is localized at the chromosome locus 1p36, which is also a frequently deleted region in HCC. As HBV infection is a major risk factor of HCC, particularly in this region and locally, we have found that the HBV viral oncoprotein HBx interacts with TAX1BP2 in co-immunofluorescence staining and co-immunoprecipitation. Taken together, these data have laid a solid foundation for TAX1BP2 to be a putative tumor suppressor gene in HCC and regulated by HBx. In this proposal, we will (1) further confirm the underexpression of TAX1BP2 in HCC with Western blotting and immunohistochemical staining and (2) also elucidate the molecular basis for the loss of TAX1BP2 expression. In addition, we will (3) determine how TAX1BP2 can suppress HCC formation and (4) the importance of its interaction with HBx. Thus findings from this study will significantly advance our understanding of TAX1BP2 in the pathogenesis of HCC and may eventually lead to the development of new drug targets and better therapeutic treatments.

Researcher : Chung LP

List of Research Outputs

Leung L.H., Tam Y.S.I., Tin P.C., Chung L.P. and Wong M.P., Src Kinase Promotes Survival and Invasion of Non-Small Cell Lung Cancer cells with Epidermal Growth Factor Receptor Abnormalities and is a Potential Candidate for Combination Targeted Therapy , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008, San Diego, California, the United State . 2008.

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Collins RJ

List of Research Outputs

Chung H.Y., Lau W.H., Chu S.S.M., Collins R.J. and Tam P.C., Carcinoid tumour of the kidney in a Chinese woman presenting with loin pain, Hong Kong Medical Journal. 2007, 13(5): 406-408.

Ng D.S.C., Chok K.S.H., Law W.L., Collins R.J. and Fan S.T., Long-term survival after resection of extrahepatic recurrence of hepatocellular carcinoma at the right colon (Letter to the Editor), International Journal of Colorectal Disease. 2007, 22(11): 1411-1412.

Researcher : Fan PYS

List of Research Outputs

Chim J.C.S., Wong M.A.T.T.H.E.W. and Fan P.Y.S., Pulmonary interstitial amyloidosis complicating multiple myeloma, Journal Clinical Oncology: official Journal of the American Society of Clinical Oncology. 2008, 26(3): 504-6.

Researcher : Guo T

List of Research Outputs

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Shen L., Au W.Y., Guo T., Wong K.Y., Wong M.L.Y., Tsuchiyama J., Yuen P.W., Kwong Y.L., Liang R.H.S. and Srivastava G., Proteasome inhibitor bortezomib-induced apoptosis in natural killer (NK)-cell leukemia and lymphoma: an in vitro and in vivo preclinical evaluation, Blood. 2007, 110(1): 469-70.

Researcher : Huang F

Project Title:	Functional conditioning of dendritic cells for DC-based tumor vaccine
Investigator(s):	Huang F, Tian L, Ng IOL
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2003

Abstract:

Dendritic cell (DC)-based tumor vaccine is a newly developed therapeutic approach for cancer treatment. It aims to promote specific immunity to cancer cells within tumor bearing individuals. Recent studies reveal that DC are not a homogenous population, and their ability to provide activation signals can vary significantly between different DC subtypes, lineages or maturity. The types and functional conditions, hence the immunogenic 'quality', of the DC employed are understandably essential. This project is to examine critically the roles of co-stimulatory molecules in DC-based tumor vaccines. The main objective is to develop immunologically active and programmable DC-tumor vaccines with high efficacy useful in cancer therapies.

Project Title:	Mechanisms for the induction and regulation of autoimmune responses by dendritic cells and T regulatory cells in systemic lupus erythematosus
Investigator(s):	Huang F, Wu AYY, Tian L
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	12/2003

Abstract:

To study the mechanisms for induction and regulation of autoimmune responses; to study effects of cell death on the induction of anti-nuclear antibody production, in normal and in autoimmune-prone mice; to test the hypothesis that uptake of necrotic or apoptotic dying cells by dendritic cells may differentially regulate autoimmune responses in vivo; to determine the roles, and mechanisms for generation, of the naturally occurring T regulatory cells in autoimmune and non-autoimmune mice.

Project Title:	Molecular characterization of dendritic cells functionally modulated by dying cells - mechanism for the induction and regulation of autoimmune responses by dendritic cells (II)
Investigator(s):	Huang F, Leung SY, Wu AYY, Tian L
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2005

Abstract:

To study the molecular mechanisms for the induction of autoimmune responses by DCs; to study the immune response profiles of DCs after uptake of dying cells; to determine how DC functional properties can be differentially modulated following uptake of apoptotic and necrotic dying cells; to determine the therapeutic potential of functionally conditioned DC for a possible long-term rectification of the autoimmune disorder.

Project Title:	Mechanistic assessment of autoimmune responses induced by dendritic cells in interleukin-10 deficient mice - implications in lupus pathogenesis
Investigator(s):	Huang F
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2007

Abstract:

To define the role of IL-10 in the control of DC-induced lupus disease development in normal and IL-10 gene-knockout mice (C57BL/6 background); to study the possible association of IL-10 gene polymorphism and lupus disease induction in the susceptible (MRL) versus resistant (C57BL/6) mouse strains; to understand the pathogenesis and genetic basis of the autoimmune disease.

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Sun Q., Kong C.T., Huang F. and Chan L.C., Aberrant Dendritic Cell Differentiation Initiated by Mll-Een Fusion Gene Does Not Requre Leukaemic Transformation., Journal of Leukocyte Biology. 2008, 83 (1): 173 - 180.

Zheng B., Lee S.S., Wong K.H., Chan K.C.W., Chan C.S., Ng F., Huang F. and Yuen K.Y., A feasibility study for detection of HIV specific cytotoxic T lymphocytes using rAAV delivery system. , 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention.. Sydney, Australia., 2007.

Researcher : Ip PPC

List of Research Outputs

Chan K.K.L., Ip P.P.C., Kwong P.W.K., Tam K.F. and Ngan H.Y.S., A combination of chemoirradiation and chemotherapy for treatment of advanced clear cell adenocarcinoma of the cervix (p 559-563) , In: John J. Kavanagh and Uziel Beller, International Journal of Gynecological Cancer. Wiley interscience, 2008, 18: 559-563.

Ip P.P.C., Lam K.W., Cheung C.L., Yeung C.W., Pun T.C., Chan K.Y.Q. and Cheung A.N.Y., Tranexamic Acid-associated Necrosis and Intralesional Thrombosis of Uterine Leiomyomas: A Clinicopathologic Study of 147 Cases Emphasizing the Importance of Drug-induced Necrosis and Early Infarcts in Leiomyomas , American Journal of Surgical Pathology. 2007, 31(8): 1215-1224.

Researcher : Khoo US

Project Title:	Association of the pro-inflammatory and anti-inflammatory cytokine gene polymorphism to breast cancer susceptibility
Investigator(s):	Khoo US, Cheung ANY, Chan YK
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2003

Abstract:

To determine whether the genetic variants of the pro-inflammatory and anti-inflammatory cytokines may (a) contribute towards breast cancer susceptibility or (b) influence prognosis; to investigate whether the joint effects of several of these alleles and/or in combination of specific environmental factors may contribute towards a stronger association.

Project Title:	Promoter polymorphisms of BRCA1: genetic association and functional study
Investigator(s):	Khoo US, Chan YK, Cheung ANY
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2004

Abstract:

To perform a case-control study to investigate the promoter polymorphisms BRCA1 gene for risk association to breast cancer; to test by Luciferase report assay whether these promoter SNPs result in functional alteration of the BRCA1 gene.

Project Title:	Role of Polymorphisms of the Inflammatory Response Genes and DC-SIGNR in Genetic Susceptibility to SARS and other infections.
Investigator(s):	Khoo US, Altmeyer RM, Chu CM, Lai ST, Lin CL, Peiris JSM, Tam PKH, Wong ATY, Yam L, Yip SP
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	12/2004

Abstract:

To determine whether (i) SNPs of the inflammatory response genes may contribute in determining susceptibilty, response to treatment and clinical outcome of SARS.(ii) Heterozygous genotypes of DC-SIGNR may influence binding affinity for viral glycopeptides, namely S1 of SARS, E2 of HCV and gp120 of HIV.

Project Title:	Infectious Diseases and Global Health: Genetic approach to the identification of host susceptibility factors and pathogen virulence determinants. Genetics of coronaviruses-associated acute respiratory disease
Investigator(s):	Khoo US
Department:	Pathology
Source(s) of Funding:	CGDN NCE Large Scale Collaborative Research Grant
Start Date:	02/2005

Abstract:

To study association of SARS-susceptibility with MHC and KIR; to carry out genetic and functional analysis of coronavirus-associated respiratory disease.

Project Title:	The role of L-SIGN (CD290L) in SARS coronavirus infection
Investigator(s):	Khoo US
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	07/2005

Abstract:

Previously we have been able to show that L_SIGN polymorphism determines the susceptibility for SARS infection. We are also to show that L-SIGN is expressed in alveolar epithelium of the healthy lung tissues, with or without co-expression of ACE2. Furthermore, L-SIGN is also expressed in the ACE2(+) alveolar epithelium of the lung of fatal SARS patients, strongly suggesting the L-SIGN may be involved in SARS pathogenesis. It has also been shown that ACE2 has been found to be expressed in the lung and small bowel enterocytes of normal individuals. In this study we intend to address specific question as below: (1) is LSIGN also expressed in the small bowel of normal individuals and SARS infected patients? (2) What is the specific role of L-SIGN in the binding and replication of SARS CoV in the setting of primary respiratory exposure/infection? - does it facilitate SARS CoV infection of permissive cells in trans or in cis, or both? (3) What is the difference in binding affinity between homozygous and heterozygous L-SIGN expression in vitro and in vivo? And how does this different account for the difference in SARS susceptibility? (4) From dbSNP (http://www.ncbi.nlm.nih.gov/SNP/), there is 12 ACE2 single nucleoride polymorphisms (SNPs) located in the coding exons, introns and promoter regions, of which four SNPs has been found with a minor allele frequency greater than 5% in our population and thus were suitable for further analysis. So we also would like to know if ACE2 SNPs is associated with different susceptibility for SARS infection.

Project Title:	The functional role of ICAM3 polymorphism in genetic susceptibility to SARS infection
Investigator(s):	Khoo US, Chan YK, Leung GM, Lin CL, Lim WWL, Peiris JSM, Sham PC, Tam PKH, Garcia-Barcelo MM, Yip SP
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	01/2006

Abstract:

To confirm our genetic association findings by a large case-control study and a family-based association study with family members as controls; to perform functional studies to investigate the role of ICAM3 polymorphisms in influencing the initiation of immune response to SARS-CoV infection.

Project Title:	Gene-based and haplotype analysis of the estrogen receptor genes for breast cancer susceptibility
Investigator(s):	Khoo US, Chan YK, Cheung ANY, Garcia-Barcelo MM, Leung GM, Sham PC, Wong MY, Zheng W
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2006

Abstract:

To identify all possible risk-conferring variations within these genes by direct sequencing of the promoter region, all exons and rrelevant exon-intron junctions on 90 unrelated individuals; to examine the pattern of linkage disequilibrium (LD) between SNPs on the genes, and hence haptotype tagging SNPs (htSNP) for risk association study; to assess the combined effect of these ER-α and ER-α htSNPs on risk of breast cancer in a large Chinese case-control population of at least 2,000 case-control pairs.

Project Title:	Promoter polymorphisms of L-SIGN in relation to host genetic susceptibility to SARS.
Investigator(s):	Khoo US, Chan YK
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	02/2006

Abstract:

Objective of study : 1. To identify the possible variants in the promoter region of L-SIGN by direct sequencing of 30 unrelated normal Chinese individuals. 2. To examine the pattern of linkage disequilibrium (LD) between all the variants identified, and hence select haptotype tagging single nucleotide polymorphism (htSNP) for risk association study. 3. To analyze these htSNPs for genetic association to SARS CoV using a large case-control study. 4. To examine for possible LD between the htSNPs and the tandem-neck repeats polymorphisms of L-SIGN previously genotyped. 5. To perform in-vitro functional studies to confirm the effect of L-SIGN promoter SNPs on the transcriptional activity of the promoter. Key issues and problems being addressed: L-SIGN or DC-SIGNR (CD209L) is a homologue of DC-SIGN (for dendritic cell-specific ICAM-3 grabbing non-integrin, CD209) and shares 77% amino acid identity with it (1). Located within 30 kb on chromosome 19p13.2-3 in a head-to-head orientation, they are thought to have arisen through a gene duplication event (2;3). The extra-cellular domain of both DC-SIGN and L-SIGN encoded by exon 4, contain tandem repeats of a highly conserved 23-amino acid sequence, followed by a C-terminal C-type carbohydrate recognition domain (CRD)(4-6). Both L-SIGN and DC-SIGN share the ability to bind to high-mannose oligosaccharides through their CRDs and serve as receptors for many viruses such as HIV, HCV and SARS Co-V. Unlike DC-SIGN, L-SIGN has considerable polymorphism in this tandem-repeat domain which encodes the extra-cellular neck region (2). This tandem-repeat segment of 3 to 9 repeats, 7 being predominant (>50%) in the general population. Homo-oligomerization through this tandem-repeat neck region is what determines its high-affinity interaction as well as ligand specificity (7). L-SIGN has been demonstrated to be a binding receptor for SARS Co-V (8). We have recently shown in a genetic association study, that individuals homozygous for L-SIGN tandem repeats are less susceptible to SARS infection, and in a series of in-vitro experiments, that L-SIGN with homozygous tandem-neck repeats had higher binding capacity for SARS-CoV, lower ability for trans infection and increased cell-association for SARS-CoV, thus playing a protective role in SARS infection (Chan et al, Nature Genetics, in press, 9). Promoter polymorphisms can cause altered binding affinity of transcription factors which may effect regulation of gene expression (10). This can give rise to differences in susceptibility to and severity of disease. A promoter polymorphism of DC-SIGN was shown to be associated with risk for parenteral acquisition of HIV-1 infection (11). This same variant was recently reported to be associated with severity of dengue disease and in-vitro study showed that it affects an Sp1-like binding site and transcriptional activity (12). We hypothesize that promoter polymorphisms may also effect the expression of L-SIGN or its association with disease. Having already shown the importance of L-SIGN as a binding receptor for SARS Co-V, and the effect of tandem-neck repeats genotype in susceptibility to SARS infection (9), it would be important to investigate whether promoter polymorphisms of L-SIGN may affect the expression of L-SIGN and thus also contribute towards disease susceptibility and/or severity for SARS. To exclude the possibility that the association with disease may be due to LD between the promoter polymorphism and the functional tandem-neck repeat homo-/heterozygosity, linkage disequilibrium (LD) between these promoter polymorphisms with tandem-neck repeat L-SIGN polymorphisms previously investigated will also be examined. Finally, in-vitro functional studies will be performed to confirm the effect of L-SIGN promoter SNPs on the promoter activity. References: 1. Soilleux EJ, Barten R, Trowsdale J. DC-SIGN; a related gene, DC-SIGNR; and CD23 form a cluster on 19p13. J Immunol 2000 September 15;165(6):2937-42. 2. Bashirova AA, et al. A dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN)-related protein is highly expressed on human liver sinusoidal endothelial cells and promotes HIV-1 infection. J Exp Med 2001 March 19;193(6):671-8. 3. Pohlmann S, et al. DC-SIGNR, a DC-SIGN homologue expressed in endothelial cells, binds to human and simian immunodeficiency viruses and activates infection in trans. Proc Natl Acad Sci U S A 2001 February 27;98(5):2670-5. 4. Feinberg H, Mitchell DA, Drickamer K, Weis WI. Structural basis for selective recognition of oligosaccharides by DC-SIGN and DC-SIGNR. Science 2001 December 7;294(5549):2163-6. 5. Guo Y, et al. Structural basis for distinct ligand-binding and targeting properties of the receptors DC-SIGN and DC-SIGNR. Nat Struct Mol Biol 2004 July;11(7):591-8. 6. Mitchell DA, Fadden AJ, Drickamer K. A novel mechanism of carbohydrate recognition by the C-type lectins DC-SIGN and DC-SIGNR. Subunit organization and binding to multivalent ligands. J Biol Chem 2001 August 3;276(31):28939-45. 7. Snyder GA, Colonna M, Sun PD. The structure of DC-SIGNR with a portion of its repeat domain lends insights to modeling of the receptor tetramer. J Mol Biol 2005 April 15;347(5):979-89. 8. Jeffers SA, Tusell SM, Gillim-Ross L, Hemmila EM, Achenbach JE, Babcock GJ et al. CD209L (L-SIGN) is a receptor for severe acute respiratory syndrome coronavirus. Proc Natl Acad Sci U S A 2004 November 2;101(44):15748-53. 9. VSF Chan, KYK Chan, YX Chen, ….US Khoo* & CL Lin* Homozygous L-SIGN (CD209L) plays a protective role in SARS coronavirus infection. Nature Genetics (in press) (*co-corresponding authors) 10. Theuns J, et al. (2003) Alzheimer-associated C allele of the promoter polymorphism -22C>T causes a critical neuron-specific decrease of presenilin 1 expression. Hum Mol Genet. 12(8), 869-877. 11. Martin MP, et al. Association of DC-SIGN promoter polymorphism with increased risk for parenteral, but not mucosal, acquisition of human immunodeficiency virus type 1 infection. J Virol. 2004; 78(24):14053-6. 12. Sakuntabhai A, et al. (2005) A variant in the CD209 promoter is associated with severity of dengue disease. Nat Genet. 37(5), 507-513. 13. Lewontin RC. The interaction of selection and linkage. II. Optimum Models. Genetics. 1964;50:757-82. 14. Wei Liu,….Khoo US, et al. The functional -969C/T promoter polymorphism in BRCA1 decreases breast cancer risk in Chinese. (Submitted to JNCI)

Project Title:	Promoter polymorphisms of DC-SIGN in relation to host genetic susceptibility to SARS infection.
Investigator(s):	Khoo US, Chan YK, Leung GM, Lin CL, Lim WWL, Peiris JSM, Sham PC, Tam PKH, Cheung ANY, Yip SP
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	06/2006

Abstract:

I. To analyze the promoter SNPs of DC-SIGN for genetic association to SARS CoV using a large case-control study and a family-based association study II. To perform in-vitro functional studies to confirm the effect of DC-SIGN promoter SNPs on promoter activity and its influence on DC-SIGN mediated transmission of SARS CoV to target cells.

Project Title:	Splice variant expression in relation to Estrogen Receptor gene expression in Chinese breast cancer
Investigator(s):	Khoo US, Chan YK, Kwong A
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	03/2007

Abstract:

Breast cancer is the commonest cause of cancer in women. Its incidence in Hong Kong Chinese has been steadily rising in the last few decades, with age-standardised rates now at 42.6 per 100,000 which although lower than Caucasian rates, is the highest rate reported in Asia. Differential gene expression profiling has the potential to substantially refine cancer prognosis beyond what is currently possible with traditional clinical and pathological indicators, thus assisting the selection of patients for the most appropriate adjuvant systemic therapies and in the classification of breast cancers based on a better understanding of cancer biology. Micro-array gene-expression profiling has been done by different research groups, with at least 2 models, namely the intrinsic-subtype model (1) and the 70-gene model (2), which have been recently validated using independent data sets (3,4). Recent data using a single data-set on 295 samples applied to five gene-expression-based models have also demonstrated high concordance rates of up to 81% agreement in their outcome predictions for individual samples (5). In spite of this however, there remains a striking lack of overlap in gene identity between gene lists provided by the different studies. The reasons for this are unknown and calls for further study. Alternative splicing (AS) is a key post-transcriptional mechanism for generating multiple protein products from a single gene, occurring in perhaps 40-60% of human genes. Alternatively spliced isoforms of a given protein can display different and even antagonistic biological functions. Mounting evidence suggests that AS is changed or becomes aberrant during the development, progression and metastasis of breast cancer (6). AS in breast cancer cells has been shown to be dependent on cell-type and culture-conditions (7). Traditional microarrays are designed to measure the total level of expression of a gene, without distinguishing between different isoforms, with probe designs being generally biased towards the 3’ end of the gene. Probes considered by standard expression-data analysis to be ‘noise’, may in fact be indicative of different patterns of regulation of multiple splice forms. Over or reduced expression of some gene isoforms may have different functions from their wild-type, and might account for differences in gene identity between the different gene-expression profile models. A genome-wide survey of human AS with exon junction microarrays, discovered AS in nearly 800 genes not previously known to be such and suggested that at least 74% of human multi-exon genes are alternatively spliced (8). Our collaborators, Merck Research Laboratories, Merck & Co., Inc., previously established a 70-gene-based prognostic classifier for breast cancers diagnosed before age 55 (9). This classifier outperformed clinical predictors in selecting out good outcome patients, thus minimizing over-treatment (2). Whilst a uniform gene expression pattern was observed for good outcome patients, patients predicted to have poor outcome had more heterogenous expression patterns. On further investigation, they were able to identify a subset of patients with high estrogen receptor expression for age which strongly predicted extremely poor outcome with distant metastases and homogenous gene expression pattern mainly of cell cycle genes (10). The reason why tumors in young patients with high ER have unique propensity to depend on proliferation associated genes is unclear which led us to hypothesize that there may be specific isoform expression pattern of genes associated with younger age onset of breast cancer. We plan to apply for RGC funding to investigate the contribution of splice-variants in on 140 Chinese breast cancer samples using a custom-made microarray which will focus on alternative splicing of the significant genes contained in their 70-gene profile previously found to be useful for prognostication of breast cancer. To obtain preliminary data in support of this study, we propose to investigate the splice-variants of some selected genes - namely the estrogen receptor alpha and beta, BRCA1 and BRCA2 and some cell cycle genes. Both quantitative expression analysis of both wild type and splice variants will be measured by Real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) using Low-density array TaqMan assay (Applied Biosystems). All breast cancer cases studied were primary invasive carcinomas treated by modified mastectomy or breast-conserving treatment, including axillary node dissection, with at least 5 to 10 year clinical follow-up and documented clinicopathological data. In a separate collaborative study with our co-investigator, we will eventually be performing microarray gene-expression profiling using the 70-gene model on these same breast cancer cases. The wild-type gene expression of these cases will thus be available for comparison with wild-type expression and splice variant data obtained in this pilot project. The objectives of this pilot study are thus as follows: Objectives: (1) To identify splice-variants of the estrogen receptor alpha and beta, BRCA1 and BRCA2 and some cell cycle genes differentially expressed in breast cancer, and investigate whether there is any correlation with age and/or clinical outcome. (2) Using high wild-type estrogen receptor levels for age as reference, to identify if there are splice variants of the same gene may be associated with opposing prognoses. Reference: 1. Perou CM, Sorlie T, Eisen MB, van de RM, Jeffrey SS, Rees CA et al. Molecular portraits of human breast tumours. Nature 2000 August 17;406(6797):747-52. 2. van d, V, He YD, van't Veer LJ, Dai H, Hart AA, Voskuil DW et al. A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med 2002 December 19;347(25):1999-2009. 3. Hu Z, Fan C, Oh DS, Marron JS, He X, Qaqish BF et al. The molecular portraits of breast tumors are conserved across microarray platforms. BMC Genomics 2006;7:96. 4. Buyse M, Loi S, van't VL, Viale G, Delorenzi M, Glas AM et al. Validation and clinical utility of a 70-gene prognostic signature for women with node-negative breast cancer. J Natl Cancer Inst 2006 September 6;98(17):1183-92. 5. Fan C, Oh DS, Wessels L, Weigelt B, Nuyten DS, Nobel AB et al. Concordance among gene-expression-based predictors for breast cancer. N Engl J Med 2006 August 10;355(6):560-9. 6. Venables JP. Aberrant and alternative splicing in cancer. Cancer Res 2004 November 1;64(21):7647-54. 7. Li C, Kato M, Shiue L, Shively JE, Ares M, Jr., Lin RJ. Cell type and culture condition-dependent alternative splicing in human breast cancer cells revealed by splicing-sensitive microarrays. Cancer Res 2006 February 15;66(4):1990-9. 8. Johnson JM, Castle J, Garrett-Engele P, Kan Z, Loerch PM, Armour CD et al. Genome-wide survey of human alternative pre-mRNA splicing with exon junction microarrays. Science 2003 December 19;302(5653):2141-4. 9. van ', V, Dai H, van d, V, He YD, Hart AA, Mao M et al. Gene expression profiling predicts clinical outcome of breast cancer. Nature 2002 January 31;415(6871):530-6. 10. Dai H, van't VL, Lamb J, He YD, Mao M, Fine BM et al. A cell proliferation signature is a marker of extremely poor outcome in a subpopulation of breast cancer patients. Cancer Res 2005 May 15;65(10):4059-66.

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Chen Y., Chan V.S.F., Zheng B., Chan Y.K., Xu X., To Y.F., Huang F.P., Khoo U.S. and Lin C.L., A novel subset of putative stem/progenitor CD34+Oct-4+ cells is the major target for SARS coronavirus in human lung., J Exp Med.. 2007, 204: 2529-36.

Cheung W.T., Lee Y.F., Ng T.W., Ching W.K., Khoo U.S., Ng M.K.P. and Wong A.S.T., CpG/CpNpG motifs in the coding region are preferred sites for mutagenesis in the breast cancer susceptibility genes, FEBS Lett. 2007, 581(24): 4668-74.

Chung P.H.Y., Wong K.K.Y., Tsoi N.S., Low L.C.K., Khoo U.S. and Tam P.K.H., A diagnostic challenge in the management of unlocalised persistent hyperinsulinaemic hypoglycaemia of infancy: a case report and review of literature, Hong Kong Journal of Paediatrics (new series). 2008, 13: 125-129.

Khoo U.S., Chan Y.K., Chan V.S. and Lin C.L., DC-SIGN and L-SIGN: the SIGNs for infection, J Mol Med. 2008.

Khoo U.S., Facutly Teaching Award, Li Ka Shing Faculty of Medicine, HKU. 2007.

Khoo U.S., Host Genetic Susceptibility to Emerging Infectious Diseases, Association of Chinese Geneticists in America and the Hong Kong Society of Medical Genetics, International Conference on Genetic and Genomic Medicine, Hong Kong. 2008.

Khoo U.S., Molecular Genetics and Pathology of Hereditary Breast and Ovarian Cancer, State of Art Lecture, 14th Hong Kong International Cancer congress and 4th International Think Tank forum, Hong Kong. 2007.

Khoo U.S., Chan Y.K., Chan V.S.F., Ching C.Y.J., Yam L., Chu C.M., Lai S.T., Wong T.Y., Tam P.K.H., Yip S.P., Leung G.M., Lin C.L. and Peiris J.S.M., Role of polymorphisms of the inflammatory response genes and DC-SIGNR in genetic susceptibility to SARS and other infections, Hong Kong Med J. 2008, 14(4)(Suppl 4): S9-12.

Kwong A., Wong L.P., Ma E.S.K., Khoo U.S. and Ford J.M., Characterization of the pathogenic mechanism of a novel BRCA2 variant in a Chinese family, Fam Cancer. 2007.

Yau T.K., Sze H., Soong I.S., Hioe F., Khoo U.S. and Lee A.W., HER2 overexpression of breast cancers in Hong Kong: prevalence and concordance between immunohistochemistry and in-situ hybridisation assays, Hong Kong Med Journal. 2008, 14(2): 130-135.

Researcher : Ko CF

List of Research Outputs

Yam J.W.P., Tse Y.T., Ko C.F., Chan L.K., Sze M.F. and Ng I.O.L., Tensin2 with growth suppression and apoptosis induction activities is underexpressed in hepatocellular carcinoma, 2008 Proceedings of the American Association for Cancer Research, San Diego, CA, USA, April. 2008.

Researcher : Ko KH

List of Research Outputs

Yeung C.M., Tan Y., Tam S., Lu L., Ko K.H., Yang P., Kung H.F. and Lin M.C., The Antidiabetic Effects of a Dry Powder of Dietary Vegetable and Fruit Mixtures in Diabetic db/db Mice, Biologics: Targets and Therapy. 2008, 2(3): 571-576.

Researcher : Kong SH

List of Research Outputs

Siu K.Y., Chan H.Y., Kong S.H., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Differential expression of folate receptor alpha and reduced folate carrier and effect of folate in ovarian cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Researcher : Lam AKY

List of Research Outputs

Chung Y.M.F., Lam A.K.Y., Luk J.M.C., Law S.Y.K., Chan K.W., Lee P.Y. and Wong J., Altered E-cadherin expression and p120 catenin localization in esophageal squamous cell carcinoma, Annals of Surgical Oncology. 2007, 14(11): 3260-3267.

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Researcher : Lam CCK

List of Research Outputs

Tse E.W.C., Cheung J.C.W., Pang A.W.K., Au W.Y., Leung A.Y.H., Lam C.C.K. and Kwong Y.L., Fludarabine, mitoxantrone and dexamethasone as first-line treatment for T-cell large granular lymphocyte leukemia, Leukemia. 2007, 21(10): 2225-2226.

Researcher : Lam QLK

List of Research Outputs

Iikuni N., Lam Q.L.K., Lu L., Matarese G. and La Cava A., Leptin and inflammation. , Current Immunology Reviews . 2008, 4: 70-79.

Lam Q.L.K., Best Abstract Award, Annual Scientific Meeting, The Hong Kong Society for Immunology. 2008.

Lam Q.L.K., Hong Kong Young Scientist Award, The Hong Kong Institute of Science. 2007.

Lam Q.L.K., International Summer Program and Internship Award, RIKEN Research Center for Allergy and Immunology, Japan. 2007.

Lam Q.L.K. and Lu L., Leptin Induces CD40 via Akt in Dendritic Cells, RIKEN Research Center for Allergy and Immunology (RCAI), Yokohama, Japan. 2007.

Lam Q.L.K. and Lu L., Leptin and LPS Signaling Pathways Converge on Akt to Upregulate CD40 Expression in Dendritic Cells, RIKEN Research Center for Allergy and Immunology, Yokohama, Japan. 2007.

Lam Q.L.K., Zheng B., Jin D., Cao X. and Lu L., Leptin induces CD40 expression through the activation of AKT in murine dendritic cells, J Biol Chem. 2007, 38: 27587-27597.

Lam Q.L.K. and Lu L., Novel Function of Leptin in Regulating Dendritic Cell Maturation and Function via the Activation of Akt Pathway, Annual Scientific Meeting, Hong Kong Society for Immunology. 2008.

Lam Q.L.K. and Lu L., Novel Function of Leptin in Regulating Dendritic Cell Maturation and Function, The 15th Annual Conference of the Hong Kong Institution of Science. 2007.

Researcher : Lau WH

List of Research Outputs

Au W.Y., Lo S.H., Law W.L., Khong P.L., Shek T.W.H., Lau W.H. and Chan E.M.C., Concomitant EBER+ve lymphomatoid papulosis and malignant transformation of colonic polyposis in a heart transplant recipient, Journal of Heart and Lung Transplantation. 2008, 27: 575-576.

Researcher : Law FBF

List of Research Outputs

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Researcher : Lee KW

List of Research Outputs

Lee K.W., Poon R.T.P., Man K., Guan X.Y., Ma S.K.Y., Liu X., Myers J.N. and Yuen P.W., Fascin over-expression is associated with aggressiveness of oral squamous cell carcinoma, Cancer Letters. 2007, 254(2): 308-315.

Ma S.K.Y., Lee K.W., Zheng B., Chan K.W. and Guan X.Y., CD133+ HCC cancer stem cells promote chemoresistance by preferential expression of the Akt/PKB survival pathway, Oncogene. 2008, 27: 1749-1758.

Researcher : Lee MY

Project Title:	Role of Cyclooxygenase-2 (COX-2) in Pro-inflammatory Response of Severe Acute Respiratory Syndrome (SARS) Coronavirus-Infected Human Primary Monocyte-derived Macrophages and the Potential Use of its Inhibitors for the Therapy of SARS
Investigator(s):	Lee MY, Peiris JSM, Cheung CY, Nicholls JM
Department:	Microbiology
Source(s) of Funding:	Small Project Funding
Start Date:	01/2007

Abstract:

Severe acute respiratory syndrome (SARS) emerged firstly in Guangdong Province, China in early 2003. The pathogen of SARS is a novel coronavirus named SARS-associated coronavirus (SARS-CoV) and is believed to be of zoonotic origin. Although the two well-known human coronaviruses 229E and OC43 cause only the common cold, several animal coronaviruses which may cause severe disease, can affect the respiratory or gastrointestinal tract or cause disseminated infections. Compared to common respiratory viral infections, SARS is unusually severe, with an overall case fatality rate of about 10%. The infection is not localized to the respiratory tract, and the causative virus is also detected in the gastrointestinal and urinary systems. Although the biological basis for the severity of SARS remained unclear, recent studies demonstrated that SARS coronavirus (SARS-CoV) can induce chemokines in macrophages in vitro and this has been suggested to contribute extensive inflammatory response and consequently to the pathogenicity of SARS-CoV (1, 2). Recent studies have demonstrated that the enzyme cyclooxygenase (COX) is involved in the host response to viral infections (3) and this enzyme is implicated as an important factor in inflammation modulation and immune response (4, 5). COX is a bifunctional enzyme, possessing the cyclooxygenase activity to convert arachidonic acid into prostaglandin G2 (PGG2), and the peroxidase activity to reduce PGG2 producing PGH2. PGH2 is then metabolized into the various bioactive prostanoids by their corresponding terminal synthases. COX exists as two different isoforms designated as COX-1 and COX-2. Although both isoforms are involved in the formation of PG endoperoxides, they are likely to have fundamentally different biological roles. COX-1 has the structural characteristic of a constitutively expressed gene and is found in most mammalian tissues. It is thought to be involved in maintaining physiological functions such as cytoprotection of the gastric mucosa, regulation of renal blood flow and control of platelet aggregation. In contrast, the expression of COX-2 is thought to be inducible and is upregulated by cytokines, bacterial lipopolysaccharide (LPS) and growth factors. Based on its distinctive expression profile, COX-2 is suggested to account for prostanoid formation in diseases such as inflammation and thus, it is the target for the design of the anti-inflammatory drug (6). Our microarray data (unpublished) showed that COX-2 gene was upregulated at 3 hours after SARS-CoV infection of human primary monocyte-derived macrophages. Importantly, SARS-CoV infected macrophages have 8.6 folds higher induction of COX-2 when compared to mock infected cells. This preliminary data suggest that COX-2 may play an important role in the cellular response to SARS-CoV infection. Based on the above issues, it would be of great interest to investigate the novel association of COX-2 and SARS-CoV and its role in the pathogenesis of SARS. Thus, the objectives of this proposed project are to investigate 1.) the involvement and role of COX-2 in pro-inflammatory response after SARS-CoV infection and 2.) the therapeutic potential of selective COX-2 inhibitors against SARS-CoV. References: 1) Cheung CY, Poon LL, Ng IH, Luk W, Sia SF, Wu MH, Chan KH, Yuen KY, Gordon S, Guan Y, Peiris JS. Cytokine responses in severe acute respiratory syndrome coronavirus-infected macrophages in vitro: possible relevance to pathogenesis. J Virol. 2005;79(12):7819-26. 2) Weiss SR, Navas-Martin S. Coronavirus pathogenesis and the emerging pathogen severe acute respiratory syndrome coronavirus. Microbiol Mol Biol Rev. 2005;69(4):635-64. 3) Carey MA, Bradbury JA, Seubert JM, Langenbach R, Zeldin DC, Germolec DR. Contrasting effects of cyclooxygenase-1 (COX-1) and COX-2 deficiency on the host response to influenza A viral infection. J Immunol. 2005;175(10):6878-84. 4) Martin JG, Suzuki M, Maghni K, Pantano R, Ramos-Barbon D, Ihaku D, Nantel F, Denis D, Hamid Q, Powell WS. The immunomodulatory actions of prostaglandin E2 on allergic airway responses in the rat. J Immunol. 2002;169(7):3963-9. 5) Carey MA, Germolec DR, Bradbury JA, Gooch RA, Moorman MP, Flake GP, Langenbach R, Zeldin DC. Accentuated T helper type 2 airway response after allergen challenge in cyclooxygenase-1-/- but not cyclooxygenase-2-/- mice. Am J Respir Crit Care Med. 2003;167(11):1509-15. 6) McAdam BF, Mardini IA, Habib A, Burke A, Lawson JA, Kapoor S, FitzGerald GA. Effect of regulated expression of human cyclooxygenase isoforms on eicosanoid and isoeicosanoid production in inflammation. J Clin Invest. 2000;105(10):1473-82.

Project Title:	Role of cyclooxygenase-2 (COX-2) influenza A (subtype H5N1) viral pathogenesis and the potential use of its inhibitors for the therapy of H5N1 disease
Investigator(s):	Lee MY, Peiris JSM, Cheung CY
Department:	Microbiology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	01/2007

Abstract:

To investigate the role of COX-2 in the pathogenesis of influenza A H5N1 disease

Researcher : Lee TYH

List of Research Outputs

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Researcher : Leung LH

List of Research Outputs

Leung L.H., Fraser M., Fiscus R.R. and Tsang B.K., Cisplatin alters nitric oxide synthase levels in human ovarian cancer cells: involvement of p53 regulation and cisplatin resistance, British Journal of Cancer. 2008, 98(11): 1803-9.

Leung L.H., Tam Y.S.I., Tin P.C., Chung L.P. and Wong M.P., Src Kinase Promotes Survival and Invasion of Non-Small Cell Lung Cancer cells with Epidermal Growth Factor Receptor Abnormalities and is a Potential Candidate for Combination Targeted Therapy , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008, San Diego, California, the United State . 2008.

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Leung RYY

List of Research Outputs

Au W.Y., Leung R.Y.Y., Mok T., Fung T.K. and Liang R.H.S., Familial occurrence of sequential B-cell lymphoma and myeloproliferative disease , Annals of Hematology. 2008, Epub.

Researcher : Leung SY

Project Title:	Identification of BRAF mutation in various stages of colorectal carcinogenesis and its relationship with KRAS mutation
Investigator(s):	Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2002

Abstract:

To examine the biological relationship of BRAF mutation with KRAS mutation. The association of mutations in these two genes with clinico-pathological features, molecular parameters and prognosis will be sought.

Project Title:	High resolution mapping of chromosomal aberrations by cDNA microarray-based comparative genomic hybridisation and their correlation with gene expression profile of gastric adenocarcinoma
Investigator(s):	Leung SY, Yuen ST, Chu KM, Lin MC
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	11/2002

Abstract:

The project attempts to achieve a high resolution mapping of the chromosomal gains and losses in primary gastric cancer samples by hybridisation of genomic DNA to a microarray containing 44,000 human cDNA clones. The changes of DNA copy number will then be correlated with gene expression profiles already available in the gastric cancer samples, generated using the same cDNA microarray. New oncogenes or tumour suppressor genes may be identified in these regions that may constitute new targets for diagnosis, prognostication and pathway specific therapeutic strategy.

Project Title:	Delineation of prognostic biomarkers in gastric cancer using cDNA microarray data, validation in independent dataset and their functional characterisation
Investigator(s):	Leung SY, Yuen ST, Chu KM, Kung H, Lin MC
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	11/2003

Abstract:

To identify a list of genes that significantly predict tumour behaviour and patient outcome in a cohort of 90 gastric adenocarcinoma patients studied by cDNA microarray; to validate the prognostic significance of the top 50 survival genes in a large independent cohort of gastric adenocarcinoma patients using a combination of real-time quantitative RT-PCR, in-situ hybridisation and immunohistochemistry performed in high-density tissue microarray; to carry out functional characterisation of the survival genes using various cell culture and animal models.

Project Title:	Detail study of relationship of BRAF and KRAS mutation with microstatellite instability in colorectal cancer
Investigator(s):	Leung SY, Yuen ST
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2003

Abstract:

To examine in detail the incidence of BRAF and KRAS mutation in a large series of MSI colorectal cancer with detail characterization of germine MMR gene mutation and MLH1 promoter methylation.

Project Title:	Genomic screening for potential tumour suppressors silenced by promoter hypermethylation in gastric adenocarcinomas using cDNA microarray
Investigator(s):	Leung SY, Yuen ST, Chu KM, Fung TWK, Lin MC, Chen X
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	01/2005

Abstract:

Gastric cancer, being the second most common cancer worldwide, continues to present with poor prognosis and obscure cause. In view of the relatively high regional incidence in Hong Kong and certain parts of China, our group has initiated an extensive study in gastric cancer using genomics approach in order to build a solid foundation for on-going research. In collaboration with Stanford University (California, U.S.A.), we have reported the gene expression profiles of 126 gastric adenocarcinomatous and non-neoplastic mucosal samples using a cDNA microarray that contains more than 30,000 unique genes. Noted from the expression profiles were around one thousand genes that were significantly down-regulated in gastric tumours compared with the non-neoplastic mucosae. Among these are many potential tumour suppressors which may contribute to the development of gastric cancer. Recent focus on epigenetics has revealed DNA methylation at the gene promoter region as a key mechanism in silencing tumour suppressor genes and hence plays an important role in carcinogenesis. The latest development of genomics approach has also allowed the systematic profiling for genes that are silenced by methylation in tumour cell lines. The data generated can then be analysed in comparison to the gene expression profiles of tumour tissue samples. This approach will lead to the identification of a comprehensive list of genes that are down-regulated in gastric tumour tissues and at the same time, re-expressed in tumour cell lines after demethylation treatment. Aims: 1. To identify genes that are induced after demethylation treatment in a large panel of gastric cancer cell lines by cDNA microarray analysis 2. To analyse the data against the expression profile database of gastric tumour tissue samples, so as to prioritise a list of genes that are down-regulated in the largest proportion of gastric tumour tissues and at the same time, induced after demethylation treatment in the largest number of gastric cancer cell lines 3. To verify the association of promoter demethylation with gene re-expression in selected gastric cancer cell lines by bisulphite genomic sequencing/Pyrosequencing and RT-PCR 4. To systematically determine the promoter methylation pattern of these target genes in gastric tumour tissues, non-neoplastic gastric mucosae and peripheral blood leucocytes, and to correlate the extent of methylation with gene expression levels in gastric tumour tissues. 5. To correlate the promoter methylation-induced silencing of specific target genes with clinico-pathological parameters, response to treatment and patient outcome

Project Title:	Systematic characterisation of genes participating in Wnt signaling pathway that regulate colon stem cell renewal and their functional significance in colon cancer pathogenesis
Investigator(s):	Leung SY, Chen X, Yuen ST, Chan TL
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	12/2006

Abstract:

To identify a set of putative Wnt target genes that are highly likely to contribute to colorectal carcinogenesis and stem cell phenotype using existing genomics databases; to validate their regulation by Wnt using quantitative RT-PCR on colon cancer cell lines with inducible blockage of Wnt signaling; to examine for variation in expression level of these validated Wnt targets in large numbers of CRCs, adenomas and normal colon mucosae using tissue microarray; to classify at a molecular level CRC using expression data of multiple validated Wnt target genes and to correlate expression levels of these validated Wnt targets with clinico-pathological parameters patient outcome and molecular genetic changes; to functionally characterize several validated Wnt targets with significant clinicopathological correlation using cell culture systems; to analyse promoters for putative Tcf binding sites in selected validated Wnt target and their confirmation using luciferase reporter assay.

List of Research Outputs

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Kosinski C., Li V.S.W., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors, Proc Natl Acad Sci U S A. 2007, 104(39): 15418-23.

Leung S.Y., Gene expression profiling of human colon tops and basal crypts - identification of BMP antagonists as intestinal stem cell niche factors, Seminar entitled “Colorectal cancer and genetics” organized by Tohoku Medical Association and Institute of Development, Aging and Cancer, Tohoku University, Japan, 9 October. 2007.

Leung S.Y., Gene expression profiling of human colon tops and basal crypts – Identification of BMP antagonists as intestinal stem cell niche factors , Laboratory of Human Carcinogenesis, Center for Cancer Research, National Cancer Institute, National Institutes of Health. Bethesda, U.S.A.. 2008.

Leung S.Y., Genetic and epigenetic mechanisms in the causation of Hereditary Nonpolyposis Colorectal Cancer Syndrome, 14th Hong Kong International Cancer Congress and 4th NFCR-AFCR-HKU International Think Tank Forum: Anti-Cancer Innovation and Global Collaboration, November 14-16, 2007, Li Ka Shing Faculty of Medicine, The University of Hong Kong. 2007.

Leung S.Y., Genetic and epigenetic mechanisms in the causation of Hereditary Nonpolyposis Colorectal Cancer Syndrome, The 14th Hong Kong International Cancer Congress and the 4th NFCR-AFCR-HKU International Think Tank Forum: Anti-Cancer Innovation and Global Collaboration, Li Ka Shing Faculty of Medicine, The University of Hong Kong. 2007.

Leung S.Y., Genetic and epigenetic mechanisms in the causation of Hereditary Nonpolyposis Colorectal Cancer Syndrome” and chairing of an “International Session entitled “Molecular Genetics and Epidemiology of Hereditary Cancer”, The 66th Annual Meeting of the Japanese Cancer Association (JCA), Yokohama, Japan, 3-5 October. 2007.

Leung S.Y., Genomics study of gastrointestinal cancers – molecular classification, prognostication and cancer biomarkers, HA COC SOM Commissioned Training Program 2007/08, Cancer: From laboratory to medicine, January 17-18. 2008.

Leung S.Y., Heritable germline methylation – implications for genetic diagnosis of Hereditary Nonpolyposis Colorectal Cancer, AACR Special Conference entitled “Cancer Epigenetics”, organized by The American Association for Cancer Research, Boston, USA, May 28 - 31. 2008.

Leung S.Y., Heritable germline methylation: implications for genetic diagnosis of Hereditary Nonpolyposis Colorectal Cancer, AACR Special Conference entitled “Cancer Epigenetics”, organized by The American Association for Cancer Research, Boston, USA. 2008.

Leung S.Y., Hyperplastic polyp – the serrated neoplasia pathway of colorectal cancer development, Asia Pacific Consensus Meeting on Colorectal Cancer Screening, Hong Kong, 15-16 September. 2007.

Leung S.Y., Microsatellite instability in Hereditary and Sporadic Colorectal Cancer – Genetic and Epigenetic Mechanisms, Association of Chinese Geneticists in America (ACGA) – Hong Kong Society of Medical Genetics (HKSMG) International Conference on Genetic and Genomic Medicine, Hong Kong, 8-11 June. 2008.

Leung S.Y., Microsatellite instability in Hereditary and Sporadic Colorectal Cancer – Genetic and Epigenetic Mechanisms, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Leung S.Y., Outstanding Researcher Award, The University of Hong Kong. 2008.

Li V.S.W., Kosinski C., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression profiling of human colon top versus basal crypts and identification of BMP antagonists as candidates for intestinal stem cell niche, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Schetter A.J., Leung S.Y., Sohn J.J., Zanetti K.A., Bowman E.D., Yanaihara N., Yuen S.T., Chan T.L., Kwong D.L., Au G.K., Liu C.G., Calin G.A., Croce C.M. and Harris C.C., MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma, JAMA. 2008, 299(4): 425-36.

Suehiro Y., Wong C.W., Chirieac L.R., Kondo Y., Shen L., Webb C.R., Chan Y.W., Chan A.S.Y., Chan T.L., Wu T.T., Rashid A., Hamanaka Y., Hinoda Y., Shannon R.L., Morris J., Issa J.P., Yuen S.T., Leung S.Y. and Hamilton S.R., Epigenetic-Genetic Interactions in the APC/WNT, RAS/RAF, and P53 Pathways in Colorectal Carcinoma, Clin Cancer Res. 2008, 14(9): 2560-9.

Sääf A.M., Halbleib J.M., Chen X., Yuen S.T., Leung S.Y., Nelson W.J. and Brown P.O., Parallels between Global Transcriptional Programs of Polarizing Caco-2 Intestinal Epithelial Cells In Vitro and Gene Expression Programs in Normal Colon and Colon Cancer, Mol Biol Cell. 2007, 18(11): 4245-60.

Researcher : Leung THY

Project Title:	Functional characterization of DLC2 in suppression of cancer cell growth and tumorigenicity in liver cancer
Investigator(s):	Leung THY, Ng IOL
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	10/2006

Abstract:

The purpose of this study is to characterize the putative tumor suppressor gene, deleted in liver cancer 2, DLC2, functionally in liver cancer (hepatocellular carcinoma, HCC), which is a prevalent cancer in Southeast Asia including Hong Kong. Although the risk factors of HCC are well established, the underlying molecular mechanisms in the development of HCC remain largely unclear. In our previous study based on sequence homology with DLC1, we have identified a homolog and a putative tumor suppressor gene, deleted in liver cancer 2, DLC2 (Ching et al., 2003). From the genetic studies on DLC2, we have demonstrated that DLC2 is frequently underexpressed in HCC and a high incidence of allelic losses using loss of heterozygosity (LOH) assay on the chromosomal locus at 13q12.3. DLC2 encodes a Rho GTPase activating protein (RhoGAP) with GAP activity specific for RhoA and Cdc42 in vitro. In addition, our recent study has demonstrated DLC2 is a RhoGAP with specific suppression of RhoA activity in vivo (Leung et al., 2005b). We have also shown the effects and roles of DLC2 in suppressing cell growth, cell migration and cellular transformation (Leung et al, 2005b). The findings were novel. However, the molecular pathways underlying the role of DLC2 remain to be elucidated. Objectives: 1. To elucidate the role of DLC2 in HCC by RNA interference approach using shRNA 2. To delineate the underlying mechanism of DLC2 in cell growth suppression 3. To determine the clinicopathological significance of DLC2 in HCC Key issues and problems being addressed: Liver cancer is a major malignancy in the world and is particularly prevalent in this region, being the second commonest fatal cancer in Southeast Asia including Hong Kong. Despite definite improvements in the outcome of patients with HCC, so far, the overall prognosis of this cancer is still unsatisfactory because of late presentation and frequent tumor recurrence after surgical resection. New adjuvant treatment modalities for HCC are much awaited. In this regard, knowledge of the molecular and cellular targets underlying the development and progression of HCC is of importance as this can provide novel opportunities for therapeutic interventions for this cancer. In studying the molecular pathways that DLC2 participates in the suppression of tumor cell growth, we found that DLC2 can suppress the activity of a ribosomal kinase p70S6K in DLC2 stably transfected clones. However, the detail mechanisms of how DLC2 inhibits the activity of p70S6K remain unclear. The upstream player of p70S6K is the mammalian target of rapamycin (mTor). mTor is one of the regulators of p70S6K and is inhibited by a drug called rapamycin. The mTor-p70S6K signaling pathway is involved in the regulation of cell growth. In addition, p70S6K also plays a role in the MAPK/ERK signaling pathway in controlling cell differentiation and growth. Delineating the linkage between DLC2, p70S6K and MAPK/ERK will be curial in studying the function of DLC2 in cancer cell development.

Researcher : Li VSW

List of Research Outputs

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Kosinski C., Li V.S.W., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors, Proc Natl Acad Sci U S A. 2007, 104(39): 15418-23.

Li V.S.W., Kosinski C., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression profiling of human colon top versus basal crypts and identification of BMP antagonists as candidates for intestinal stem cell niche, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Researcher : Liao X

List of Research Outputs

Chan K.K.L., Wei N., Liu S., Liao X., Cheung A.N.Y. and Ngan H.Y.S., Estrogen receptor subtypes in ovarian cancer: a clinical correlation, Obstet Gynecol. 2008, 111: 144-151.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Over-expression of Prostatic Stem Cell Antigen (PSCA) is associated with Gestational Trophoblastic neoplasia. , Histopathology. 2007, 52: 167-174.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Overexpression of prostate stem cell antigen is associated with gestational trophoblastic neoplasia, Histopathology. 2008, 52(2): 167-174.

Liao X., Siu K.Y., Au W.H., Wong E.S.Y., Ngan H.Y.S. and Cheung A.N.Y., Aberrant Activation of Hedgehog Signaling Pathway in Ovarian Cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Researcher : Ling GS

List of Research Outputs

Chen Y.X., Man K., Ling G.S., Chen Y., Sun B., Cheng Q., Wong O.H., Lo C.K., Ng I.O.L., Chan L.C., Lau G., Lin S.C.L., Huang F. and Huang F.P., A crucial role for dendritic cell (DC) IL-10 in inhibiting successful DC-based immunotherapy: superior antitumor immunity against hepatocellular carcinoma evoked by DC devoid of IL-10, Journal of Immunology. 2007, 179(9): 6009-6015.

Researcher : Liu W

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Researcher : Lo CK

List of Research Outputs

Chen Y.X., Man K., Ling G.S., Chen Y., Sun B., Cheng Q., Wong O.H., Lo C.K., Ng I.O.L., Chan L.C., Lau G., Lin S.C.L., Huang F. and Huang F.P., A crucial role for dendritic cell (DC) IL-10 in inhibiting successful DC-based immunotherapy: superior antitumor immunity against hepatocellular carcinoma evoked by DC devoid of IL-10, Journal of Immunology. 2007, 179(9): 6009-6015.

Researcher : Lo MWS

List of Research Outputs

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Researcher : Loong F

List of Research Outputs

Chim J.C.S., Wong K.Y., Loong F., Lam W.W. and Srivastava G., Frequent epigenetic inactivation of Rb1 in addition to p15 and p16 in mantle cell and follicular lymphoma, Hum Pathol. 2007, 38(12): 1849-57.

Chim J.C.S., Wong K.Y., Loong F., Lam W.W. and Srivastava G., Frequent epigenetic inactivation of Rb1 in addition to p15 and p16 in mantle cell and follicular lymphoma, Human Pathology. 2007, 38(12): 1849-57.

Hwang Y.Y., Chim J.C.S., Chan G., Loong F. and Yau T., Breast and pelvic masses in a myeloma patient, Annals of Hematology. Springer Berlin / Heidelberg, 2008.

Researcher : Lu L

Project Title:	B cell apoptosis and its regulation in autoimmunity
Investigator(s):	Lu L
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	09/2003

Abstract:

To study the regulation of B cell development and survival by BLyS and APRIL in mouse bone marrow; to examine B cell apoptosis and its regulation in the development of CIA as a model of RA; to determine if BLyS and/or APRIL participate(s) in the pathogenesis of CIA.

Project Title:	Natural killer cells and the pathogenesis of autoimmunity
Investigator(s):	Lu L
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	10/2004

Abstract:

To study : (1) NK cell development during the pathogenesis of autoimmune arthritis. (2) NK cell interaction with T and B cells under autoimmune conditions. (3) role of NK cells in the development of autoimmune arthritis.

Project Title:	Functional interactions of dendritic cells and B cells in autoimmunity
Investigator(s):	Lu L
Department:	Pathology
Source(s) of Funding:	NSFC/RGC Joint Research Scheme
Start Date:	03/2005

Abstract:

To study: (1) roles of BLyS in regulating DCs and B cell functions in autoimmunity, (2) roles of heat-shock proteins in regulating functions of DCs and B cells, (3) roles of HSPs in regulating autoimmune arthritis development.

Project Title:	Role of leptin receptor signaling in the maturation and function of dendritic cells
Investigator(s):	Lu L
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	08/2006

Abstract:

Leptin, originally discovered as a hormone produced primarily in adipose cells, plays a critical role in regulating nutrient intake and metabolism. There is increasing evidence that leptin acts as a growth factor by stimulating proliferation and inhibiting apoptosis in human and murine T lymphocytes (1-3). Several studies have revealed an important role for leptin in regulating lymphopoiesis and hematopoeisis. For example, leptin has been shown to increase the production of a variety of cytokines in rodent T cells and monocytes/macrophages (1,4-6). Moreover, leptin is involved in modulating immune responses towards the Th1 phenotype and suppressing the Th2 effect by stimulating CD4+ T lymphocyte proliferation (1,7). In relation to the important function of leptin in immune response, the leptin receptor expression has been found not only in the central nervous system but also in the peripheral lymphoid tissues and hematopoietic stem cells (1,8-10). Encoded by the diabete (db) gene, the leptin receptor (Ob-R) is a member of class I cytokine receptors, which has signaling capability of IL-6 type cytokine receptors. Ob-R mRNA gives rise to six different forms of the receptor by alternative splicing but only the long isoform (Ob-Rb) has been recognized to be of prime importance in leptin-mediated signaling. Early studies on db/db mice, in which the leptin receptor Ob-Rb that transduces signals through leptin binding is truncated, have revealed that B and T cell development and maturation are severely affected with reduced numbers of lymphocytes in the peripheral lymphoid organs (8). Dendritic cells (DCs) play a pivotal role as professional APCs due to its high potency and versatility in immune function. We have recently identified the microenvironmental factors that can affect DC function (11,12). Moreover, our pilot studies have demonstrated that DCs express surface leptin receptor as analyzed by quantitative PCR and immunofluorescence microscopy. Although the role of leptin in regulating immune functions of lymphocytes and macrophage has been widely studied, whether and to what extent leptin signaling modulates DC function remain largely unclear. To better understand the physiological function of leptin signaling in DC development, this project will focus on studying DC maturation and function in leptin-receptor deficient db/db mice. Objectives of this proposal:1. To examine the phenotype and costimulatory molecule expression on DCs derived from db/db BM cultures. 2. To evaluate the capacity of db/db DCs in stimulating allogeneic T cell proliferation.3. To determine the profile of cytokine production by db/db DCs.4. To elucidate the signaling transduction pathways underlying the maturation and function of db/db DCs.References1. Lord, G. M., Matarese, G., Howard, J. K., Baker, R. J., Bloom, S. R., and Lechler, R. I. 1998. Leptin modulates the T-cell immune response and reverses starvation-induced immunosuppression. Nature 394:897-901.2. Howard, J. K., Lord, G. M., Matarese, G., Vendetti, S., Ghatei, M. A., Ritter, M. A., Lechler, R. I., and Bloom, S. R. 1999. Leptin protects mice from starvation-induced lymphoid atrophy and increases thymic cellularity in ob/ob mice. J Clin Invest 104:1051-9.3. Fujita, Y., Murakami, M., Ogawa, Y., Masuzaki, H., Tanaka, M., Ozaki, S., Nakao, K., and Mimori, T. 2002. Leptin inhibits stress-induced apoptosis of T lymphocytes. Clin Exp Immunol 128:21-6.4. Faggioni, R., Feingold, K. R., and Grunfeld, C. 2001. Leptin regulation of the immune response and the immunodeficiency of malnutrition. Faseb J 15:2565-71.5. Loffreda, S., Yang, S. Q., Lin, H. Z., Karp, C. L., Brengman, M. L., Wang, D. J., Klein, A. S., Bulkley, G. B., Bao, C., and Diehl, A. M. 1998. Leptin regulates proinflammatory immune responses. Faseb J 12:57-65.6. Santos-Alvarez, J., Goberna, R., and Sanchez-Margalet, V. 1999. Human leptin stimulates proliferation and activation of human circulating monocytes. Cell Immunol 194:6-11.7. Shigemura, N., Ohta, R., Kusakabe, Y., Miura, H., Hino, A., Koyano, K., Nakashima, K., and Ninomiya, Y. 2004. Leptin modulates behavioral responses to sweet substances by influencing peripheral taste structures. Endocrinology 145:839-47.8. Bennett, B. D., Solar, G. P., Yuan, J. Q., Mathias, J., Thomas, G. R., and Matthews, W. 1996. A role for leptin and its cognate receptor in hematopoiesis. Curr Biol 6:1170-80.9. Nakata, M., Yada, T., Soejima, N., and Maruyama, I. 1999. Leptin promotes aggregation of human platelets via the long form of its receptor. Diabetes 48:426-9.10. Cioffi, J. A., Shafer, A. W., Zupancic, T. J., Smith-Gbur, J., Platika, D., and Snodgrass, H. R. 1996. Novel B219/OB receptor isoforms: possible role of leptin in hematopoiesis and reproduction. Nat Med 2:585-9.11. Qu X., Yang M.X., Kong B.H., Qi L., Lam Q.L.K., Yan S., Zhang M. and Lu L., Hypoxia inhibits the migratory capacity of human monocyte-derived dendritic cells. Immunology and Cell Biology, 2005, 83: 668-73. 12. Yang M.X., Qu X., Kong B.H., Lam Q.L.K., Shao Q., Deng B.P., Ko K.H. and Lu L., Membrane type 1-matrix metalloproteinase is involved in the migration of human monocyte-derived dendritic cells. Immunology and Cell Biology (in press)

Project Title:	Innovative development of a gene-targeted therapy for rheumatoid arthritis
Investigator(s):	Lu L, Ko KH
Department:	Pathology
Source(s) of Funding:	Innovation and Technology Support Programme
Start Date:	09/2006

Abstract:

1. To characterize the immune mechanisms underlying the pathogenic role of BAFF in the development of autoimmune arthritis. 2. To construct a recombinant lentiviral expression system to deliver small hairpin RNAs for BAFF-gene silencing. 3. To evaluate the efficacy and safety of in vivo application of the recombinant lentiviral vector expressing short-hairpin RNA for BAFF gene-silencing in the treatment of autoimmune arthritis.

Project Title:	Plasma cell development and its regulation in autoimmunity
Investigator(s):	Lu L
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2007

Abstract:

To study PC generation and functional characteristics upon immunization; to examine the regulatory mechanisms underlying PC migration and survival; to characterize the development of PCs during the progression of autoimmune arthritis; to study the molecular mechanism involved in regulating PC development in autoimmune arthritis.

List of Research Outputs

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Iikuni N., Lam Q.L.K., Lu L., Matarese G. and La Cava A., Leptin and inflammation. , Current Immunology Reviews . 2008, 4: 70-79.

Lam Q.L.K. and Lu L., Leptin Induces CD40 via Akt in Dendritic Cells, RIKEN Research Center for Allergy and Immunology (RCAI), Yokohama, Japan. 2007.

Lam Q.L.K. and Lu L., Leptin and LPS Signaling Pathways Converge on Akt to Upregulate CD40 Expression in Dendritic Cells, RIKEN Research Center for Allergy and Immunology, Yokohama, Japan. 2007.

Lam Q.L.K., Zheng B., Jin D., Cao X. and Lu L., Leptin induces CD40 expression through the activation of AKT in murine dendritic cells, J Biol Chem. 2007, 38: 27587-27597.

Lam Q.L.K. and Lu L., Novel Function of Leptin in Regulating Dendritic Cell Maturation and Function via the Activation of Akt Pathway, Annual Scientific Meeting, Hong Kong Society for Immunology. 2008.

Lam Q.L.K. and Lu L., Novel Function of Leptin in Regulating Dendritic Cell Maturation and Function, The 15th Annual Conference of the Hong Kong Institution of Science. 2007.

Lu L., B cell maturation and its dysregulation in autoimmunity, 95th Annual Conference of American Association of Immunologists, San Diego, USA. 2008.

Lu L., B cell maturation and its dysregulation in autoimmunity, Guest Society Symposium, 95th Annual Conference of American Association of Immunologists, San Diego, USA. 2008.

Lu L., Role of B cell activating factor in the pathogenesis of autoimmune arthritis, International Conference on Immunology 2007 & The First World Conference of Chinese Immunologists, Shanghai, China, July 12-15, 2007. 2007.

Sun L.Y., Zhou K.X., Feng X.B., Zhang H.Y., Ding X.Q., Jin O., Lu L., Lau W.C.S., Hou Y.Y. and Fan L.M., Abnormal Surface Markers Expression on Bone Marrow CD34+ cells and Correlation with Disease Activity in Patients with Systemic Lupus Erythematosus, Clinical Rheumatology. Belgium, Acta Medica Belgica, 2007, 26: 2073-2079.

Sun L.Y., Zhang H.Y., Feng X.B., Hou Y., Lu L. and Fan L.M., Abnormality of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus, Lupus. 2007, 16(2): 121-8.

Yeung C.M., Tan Y., Tam S., Lu L., Ko K.H., Yang P., Kung H.F. and Lin M.C., The Antidiabetic Effects of a Dry Powder of Dietary Vegetable and Fruit Mixtures in Diabetic db/db Mice, Biologics: Targets and Therapy. 2008, 2(3): 571-576.

Zhou J., Huang J., Poon K.M., Chen D., Chan C.S., Ng F., Guan X.Y., Watt R.M., Lu L., Yuen K.Y. and Zheng B., A mechanism underlying susceptibility to HBV infection: HBV down-regulate transferase-1 type I interferon receptor 1 via suppressing poly(ADP-ribose), 2007 International Meeting: The Molecular Biology of Hepatitis B Viruses.. 2007.

Researcher : Ma ESK

List of Research Outputs

Chung B.H.Y., Ma E.S.K., Khong P.L. and Chan G.C.F., Inherited Thrombophilic factors do not increase central venous catheter blockage in Chinese children with malignancy, Pediatr Blood & Cancer (Epub June 2008). 2008.

Gibney G.T., Panhuysen C.I.M., So J.C.C., Ma E.S.K., Ha S.Y., Li C.K., Lee A.C.W., Li C.K., Yuen H.L., Lau Y.L., Johnson D.M., Farrell J.J., Bisbee A.B., Farrer L.A., Steinberg M.H., Chan L.C. and Chui D.H.K., Variation and heritability of Hb F and F-cells among beta-thalassemia heterozygotes in Hong Kong., Am J Hematol. 2008, 458-464.

Kwong A., Wong L.P., Ma E.S.K., Khoo U.S. and Ford J.M., Characterization of the pathogenic mechanism of a novel BRCA2 variant in a Chinese family, Fam Cancer. 2007.

Lee A.C., Ma E.S.K., Chan A.Y., Szeto S.C. and Chan L.C., Double heterozygosity for Hb New York [beta 113 GTG-->GAG; VAL-->GLU] and beta degrees-thalassemia mutations manifests as a thalassemia trait, Pediatr Hematol Oncol. 2008, 25(3): 227-31.

So J.C.C., Ma E.S.K., Wan T.S.K., Yip S.F. and Chan L.C., Clinicopathological features of unbalanced translocation Der(1;7)(q10;p10) in myeloid neoplasms, Leuk Res. 2008, 32(6): 1000-1001.

Researcher : Ma SKY

List of Research Outputs

Lee K.W., Poon R.T.P., Man K., Guan X.Y., Ma S.K.Y., Liu X., Myers J.N. and Yuen P.W., Fascin over-expression is associated with aggressiveness of oral squamous cell carcinoma, Cancer Letters. 2007, 254(2): 308-315.

Lee K.W., Poon R.T.P., Wo Y.H., Ma S.K.Y., Guan X.Y., Myers J.N., Altevogt P. and Yuen P.W., Lupeol suppresses cisplatin induced NFkB activation in HNSCC cells and inhibits local invasion and nodal metastasis in orthotopic nude mouse model of tongue squamous cell carcinoma, Cancer Research. 2007, 67(18): 8800-8809.

Ma S.K.Y., Lee K.Y., Zheng B. and Chan K.W., CD133(+) HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway., Oncogene. 2008, 27: 1749-1758.

Ma S.K.Y., Lee K.W., Zheng B., Chan K.W. and Guan X.Y., CD133+ HCC cancer stem cells promote chemoresistance by preferential expression of the Akt/PKB survival pathway, Oncogene. 2008, 27: 1749-1758.

Ma S.K.Y., Guan X.Y., Lee K.W. and Chan K.W., Clinicopathological significance of missing in metastasis B expression in hepatocellular carcinoma, Hum Pathol. 2007, 38(8): 1201-6.

Ma S.K.Y., Hu L., Huang X.H., Cao L.Q., Chan K.W., Wang Q. and Guan X.Y., Establishment and characterization of a human cholangiocarcinoma cell line., Oncology Report. 2007, 18: 1195-1200.

Ma S.K.Y. and Guan X.Y., Identification and characterization of liver cancer stem cells, Hong Kong - Shanghai International Liver Congress. 2008.

Ma S.K.Y., Mary Sun Medical Scholarship, The University of Hong Kong. 2007.

Researcher : Ma SKY

List of Research Outputs

Ma S.K.Y., Lee K.Y., Zheng B. and Chan K.W., CD133(+) HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway., Oncogene. 2008, 27: 1749-1758.

Ma S.K.Y., Guan X.Y., Lee K.W. and Chan K.W., Clinicopathological significance of missing in metastasis B expression in hepatocellular carcinoma, Hum Pathol. 2007, 38(8): 1201-6.

Ma S.K.Y., Hu L., Huang X.H., Cao L.Q., Chan K.W., Wang Q. and Guan X.Y., Establishment and characterization of a human cholangiocarcinoma cell line., Oncology Report. 2007, 18: 1195-1200.

Ma S.K.Y. and Guan X.Y., Identification and characterization of liver cancer stem cells, Hong Kong - Shanghai International Liver Congress. 2008.

Ma S.K.Y., Mary Sun Medical Scholarship, The University of Hong Kong. 2007.

Researcher : Mak CM

List of Research Outputs

Fung C.W., Blau N., Siu S., Mak C.M., Cheung P.T., Tam S. and Wong V.C.N., A new presentation of 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency Dopa Responsive Dystonia (DRD), 12th National Child Neurology Congress (第十二届全國小兒神經學術會議), Wuyishan, China, 27-29 October, 2007.

Fung C.W., Siu S., Mak C.M., Poon G.W.K., Wong K.Y., Cheung P.T., Low L.C.K., Tam S. and Wong V.C.N., A rare cause of hepatosplenomegaly transaldolase deficiency, 12th National Child Neurology Congress (第十二届全國小兒神經學術會議), Wuyishan, China, 27-29 October, 2007.

Fung C.W., Siu S., Mak C.M., Poon G.W.K., Wong K.Y., Cheung P.T., Low L.C.K., Tam S. and Wong V.C.N., A rare cause of hepatosplenomegaly transaldolase deficiency, A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Cheung P.T., Tam S. and Wong V.C.N., A rare presentation of 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency - Dopa Responsive Dystronia (DRD), A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Cheung P.T., Mak C.M., Siu S., Tam S. and Wong V.C.N., A study of cerebrospinal fluid neurotransmitters assay in children with undiagnosed neurological diseases in Hong Kong, Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society and HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Tam S. and Wong V.C.N., An atypical presentation of cerebral folate deficiency, 12th National Child Neurology Congress (第十二届全國小兒神經學術會議), Wuyishan, China, 27-29 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Tam S. and Wong V.C.N., An atypical presentation of cerebral folate deficiency, A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Tam S. and Wong V.C.N., An atypical presentation of cerebral folate deficiency, Annual Symposium of Society for the Study of Inborn Errors of Metabolism, Hamburg, 4-7 September 2007.

Luk H.M., Fung C.W., Mak C.M., Lam C.W., Tam S. and Wong V.C.N., Severe myoclonic epilepsy in infancy (SMEI) or Dravet Syndrome - case report with SCN1A mutation in Hong Kong, A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Researcher : Ng IOL

Project Title:	Characterization of T-cadherin in hepatocellular carcinoma
Investigator(s):	Ng IOL, Chan DW
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2003

Abstract:

To evaluate the expression of T-cadherin at mRNA and protein levels in human HCCs and in HCC cell lines; to investigate the factors which induce expression of T-cadherin in HCC.

Project Title:	Functional characterization of DLC1 gene, a novel tumour suppressor gene frequently deleted in liver cancer
Investigator(s):	Ng IOL, Yau TO, Sham MH, Jin D, Ching YP
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2005

Abstract:

To develop and characterize DLC-null mice using gene targeting technology; to delineate the interaction between DLC1 and the downstream effector, ROCK in HCC cell lines and human HCCs.

Project Title:	Genome-wide methylation screening in search for novel tumour suppressor genes in liver cancer
Investigator(s):	Ng IOL, Ching YP, Wong CM
Department:	Pathology
Source(s) of Funding:	Michael Kadoorie Cancer Genetics Research Fund
Start Date:	01/2005

Abstract:

To characterize HCC-specific hypermethylation profile in HCC cells using microarray approach; to identify novel tumour suppressor genes in HCC cells; to delineate epigenetic and genetic alterations of selected putative tumour suppressor genes in human HCC; to characterize tumour suppressor activities of selected novel tumour suppressor genes.

Project Title:	Characterization of DLC1 gene, a novel tumour suppressor gene frequently deleted in liver cancer
Investigator(s):	Ng IOL, Jin D, Yam JWP, Ching YP
Department:	Pathology
Source(s) of Funding:	Michael Kadoorie Cancer Genetics Research Fund
Start Date:	01/2005

Abstract:

To search for binding partners of DLC1; to delineate the interaction between DLC1 and the downstream effector, ROCK, in HCC cell lines and human HCCs; to assess the clinicopathologic significance of ROCK and its relationship with DLC1 in HCCs.

Project Title:	Characterization of the dishevelled gene in Wnt/[beta]-catenin signaling in liver cancer
Investigator(s):	Ng IOL, Yam JWP, Chan DW
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	01/2005

Abstract:

The main objectives of this project are to: 1) characterize the expression of Dvl, HDPR1, and Pk1 in HCC and evaluate their clinicopathological significance, 2) delineate the genetic and epignetic alterations of Dvl, HDPR1 and Pk1 in HCC, 3) assess the biochemical properties of Dvl in HCC cells, and 4) investigate the functions of Dvl, HDPR1, and Pk1 in Wnt signaling in HCC development.

Project Title:	A study of molecular mechanisms and characterizing novel genes in liver cancer
Investigator(s):	Ng IOL
Department:	Pathology
Source(s) of Funding:	Senior Medical Research Fellowships
Start Date:	09/2005

Abstract:

To investigate the genetic basis of genome instability in liver cancer in relation to competence of mitotic checkpoints, and hepatitis B virus. Different approaches will be used to study the functional aspects as well as clinicopathological and prognostic significance of these genetic changes, in order to shed light for new treatment modalities.

Project Title:	LRP6 coreceptor upregulation in liver cancer
Investigator(s):	Ng IOL
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	04/2006

Abstract:

The purpose of this study is to characterize LRP6 (low-density lipoprotein receptor related protein) genetically and functionally in liver cancer, which is a major cancer in Southeast Asia and Hong Kong. Activation of Wnt/β-catenin pathways is involved in the formation and progression of various cancers. However, the role of Wnt signaling has not been extensively examined in liver cancer (hepatocellular carcinoma, HCC). Our previous studies have already documented the importance of players of this pathway, such as HDPR1 and β-catenin genes, in HCC development. Here we hypothesize that LRP6, a newly identified co-receptor of Wnt/β-catenin canonical signaling pathway, is a putative oncogene in HCC and its dysregulation plays a role in hepatocarcinogenesis. We will examine the mechanism of LRP6 upregulation in human HCC and the molecular pathways by which LRP6 exerts its effects. Our work should shed light on how LRP6 regulates the Wnt canonical signaling and how LRP6 overexpression leads to HCC formation. Objectives to be achieved are: 1. To evaluate the expression profile, genetic alterations, and clinical significance of LRP6 in HCC 2. To characterize the tumorigenic activity of LRP6 in HCC Key issues and problems being addressed: - Liver cancer is a major malignancy in the world and is particularly prevalent in this region, being the second commonest fatal cancer in Southeast Asia including Hong Kong. Despite definite improvements in the outcome of patients with HCC, so far, the overall prognosis of this cancer is still unsatisfactory because of late presentation and frequent tumor recurrence after surgical resection. New adjuvant treatment modalities for HCC are much awaited. In this regard, knowledge of the molecular and cellular targets underlying the development and progression of HCC is of importance as this can provide novel opportunities for therapeutic interventions for this cancer. - Activation of canonical Wnt signaling through β-catenin/T cell factor complexes is a key event in embryonic development and is closely linked to cancer formation. β-catenin activation is a pivotal event in this pathway. From others’ and our previous results, β-catenin overexpression in human HCC is frequent (50-68%) but somatic mutations of β-catenin occur in much lower frequencies. Evidence has shown that factors other than β-catenin mutation contribute to β-catenin activation. The Wnt signal requires two membrane receptors for the Wnt ligands, Frizzled and LRP, to form a Wnt-induced Frizzled-LRP complex. LRP5 and LRP6, identified in 1998, are essential co-receptors of Wnt and specifically function in the canonical pathway. The data derived in recent studies have suggested that LRP6 is a putative oncogene in cancers. However, the pathogenic roles of LRP5 and LRP6 have not been explored in HCC. - As part of our continual study on the role of Wnt/β-catenin in HCC, we have found in our preliminary study that LRP6 expression was frequently and significantly upregulated in human HCCs, whereas LRP5 expression was not. This significant upregulated expression in HCC suggests that dysregulation of LRP6 is implicated in hepatocarcinogenesis. However, there is minimal information available on the expression profile, tumorigenic potential and functional characteristics of LRP6. Further investigations are warranted.

Project Title:	Characterization of the roles of hepatitis B virus X protein in liver cancer
Investigator(s):	Ng IOL, Chan DW
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	09/2006

Abstract:

To investigate the alterations of HBx cell targets in HCC cells with knockdown of HBx by RNA interference (RNAi); to delineate the role of HBx in deregulation of mitotic checkpoint control; to characterize the cellular effects of natural HBx mutants in HCC cells.

Project Title:	Characterization of regulatory molecules of Wnt/beta-catenin signaling in liver cancer
Investigator(s):	Ng IOL
Department:	Pathology
Source(s) of Funding:	NSFC/RGC Joint Research Scheme
Start Date:	01/2007

Abstract:

To analyze the expression patterns and activities of Dishevelled (Dv1) and DICKKOPf-1 (DKK1) in the Wnt/beta;-catenin signaling pathway in HCC tissues and cells; to functionally characterize these proteins of Wnt/β-catenin signaling pathway in HCC cells; to evaluate the clinicopathological and prognostic significance of these genes in human HCC.

Project Title:	Significance of LRP6 coreceptor upregulation in the aberrant activation of Wnt signaling in liver cancer
Investigator(s):	Ng IOL, Yam JWP, Ching YP, Yau TO
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2007

Abstract:

To evaluate the expression profile, genetic alterations, and clinical significance of LRP6 in HCC; to characterize the tumorigenic activity of LRP6 in HCC; to generate and characterize mouse models with overexpression of LRP6 in HCC cells.

Project Title:	Functional characterization of the mitotic checkpoint protein MAD1 in liver cancer
Investigator(s):	Ng IOL
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	03/2007

Abstract:

Liver cancer (hepatocellular carcinoma, HCC) is a major malignancy worldwide and particularly prevalent in this region. In HCC, recurrent chromosomal abnormalities are common and these frequent gains and losses of chromosomes suggest that chromosomal instability significantly contributes to liver cancer formation. One major mechanism for chromosomal instability is through loss of mitotic checkpoint, which ensures orderly and faithful progression of cell division. MAD1 is a key protein in mitotic checkpoint control first identified by us, but its role in HCC is unclear. Recently, we have identified a novel, alternatively splicing isoform of MAD1 (MAD1-beta). Unlike MAD1, which localizes in nucleus, MAD1-beta localizes in cytoplasm. Our preliminary data suggest that MAD1-beta may bring MAD2 out of the nucleus, thus inhibiting mitotic checkpoint and promoting carcinogenesis. We hypothesize that MAD1 is dysregulated in HCC and both MAD1 and MAD1-beta bear pathogenetic and clinicopathological significance. We aim to systemically compare the characteristics of MAD1 and MAD1-beta and evaluate the expression and clinicopathological significance of MAD1/MAD1beta and their correlation with HBx in HCC. Our work will significantly advance our understanding of HCC development and may provide insight for establishment of new effective therapy targeting the mitotic checkpoint.

Project Title:	Molecular pathology of liver cancer - a multidisciplinary study
Investigator(s):	Ng IOL, Cheung ST, Poon RTP, Ching YP, Guan XY, Jin D, Fan ST, Huang J, Lai CL
Department:	Pathology
Source(s) of Funding:	Central Allocation Vote - Group Research Project
Start Date:	04/2007

Abstract:

1) To elucidate the genetic and molecular alterations in multistep hepatocarcinogenesis – We will use a comprehensive approach to perform genome-wide analysis with HCC tissues and cells as well as animal models. We will determine microRNA expression profile and also characterize the role of hepatitis B virus (HBV) in different disease stages of HCC. 2) To characterize Rho/ROCK/PTEN/AKT signaling pathway in this multistep hepatocarcinogenesis - We will define the critical events in this major oncogenic pathway that have significant impact on HCC development and progression. Particularly, we will delineate the role of this pathway in angiogenesis.

List of Research Outputs

Chen Y.X., Man K., Ling G.S., Chen Y., Sun B., Cheng Q., Wong O.H., Lo C.K., Ng I.O.L., Chan L.C., Lau G., Lin S.C.L., Huang F. and Huang F.P., A crucial role for dendritic cell (DC) IL-10 in inhibiting successful DC-based immunotherapy: superior antitumor immunity against hepatocellular carcinoma evoked by DC devoid of IL-10, Journal of Immunology. 2007, 179(9): 6009-6015.

Cheung S.T., Fan S.T., Lee Y.T., Chow J.P.H., Ng I.O.L., Fong D.Y.T. and Lo C.M., Albumin mRNA in plasma predicts post-transplant recurrence of patients with hepatocellular carcinoma, Transplantation. 2008, 85(1): 81-87.

Ho J.C.Y., Cheung S.T., Poon W.S., Lee Y.T., Ng I.O.L. and Fan S.T., Down-regulation of retinol binding protein 5 is associated with aggressive tumor features in hepatocellular carcinoma, Journal of Cancer Research and Clinical Oncology. 2007, 133(12): 929-936.

Leung T.H.Y., Yam J.W.P. and Ng I.O.L., STARD13 (StAR-related lipid transfer (START) domain containing 13). , Atlas Genet Cytogenet Oncol Haematol.. 2007.

Low S.C.S., Peh W.C.G., Muttarak M., Cheung H.S. and Ng I.O.L., Imaging features of hepatic angiomyolipomas, Journal of Medical Imaging and Radiation Oncology. 2008, 52(2): 118-123.

Mak G.W.Y., Leong V.Y.L., Yau T.O., Ng I.O.L. and Ching Y.P., Functional characterization of C53 and its possible roles in tumorigenesis, Oncogenes and Human Cancer- The next 25 years / Nature-Centro Nacional de Investigaciones Oncologicas (CNIO) Conference . 2007.

Man K., Lo C.M., Xiao J., Ng T.P., Sun B., Ng I.O.L., Cheng Q., Sun K.W. and Fan S.T., The significance of acute phase small-for-size graft injury on tumor growth and invasiveness after liver transplantation, Annals of Surgery. 2008, 247(6): 1049-1057.

Ng I.O.L., Asian Journal of Oral and Maxillofacial Surgery. 2007.

Ng I.O.L., Asian Journal of Surgery. 2007.

Ng I.O.L., Journal of Clinical Oncology (Chinese version). 2007.

Ng I.O.L., Journal of Gastroenterology and Hepatology. 2007.

Ng I.O.L., Liver International, Associate Editor. 2007.

Ng I.O.L., Loke Yew Endowed Professorship in Pathology, The University of Hong Kong. 2008.

Ng I.O.L., Molecular characterization of human hepatocellular carcinoma – Tumor suppressor genes, 2008.

Ng I.O.L., Molecular characterization of human hepatocellular carcinoma: Tumor suppressor genes, San Diego, USA, American Association for Cancer Research 2008 Annual Meeting, 2008.

Sun K.W., Ng T.P., Sun B.S., Ho J.W.Y., Lee K.W., Ng I.O.L., Poon R.T.P., Lo C.M., Liu C.L., Man K. and Fan S.T., The significance of proline-rich tyrosine kinase2 (Pyk2) on hepatocellular carcinoma progression and recurrence, British Journal of Cancer. 2007, 97(1): 50-57.

Sze M.F. and Ng I.O.L., Application of an inducible gene expression system to functionally study hepatitis B virus X protein, Health Research Symposium 2007: Building bridges between research, practice & policy, September 29. 2007.

Wong C.L. and Ng I.O.L., Rho-kinase 2 (ROCK2) is involved in the progression and invasion of hepatocellular carcinoma cells by regulating actomyosin contractility, The Hong Kong International Cancer Congress. 2007.

Wong C.M. and Ng I.O.L., Frequent overexpression of SUV39H1 in human hepatocellular carcinoma, American Association for Cancer Research Special Meeting: Cancer Epigenetics, Boston, MA, USA, May. 2008.

Wong C.M. and Ng I.O.L., Molecular pathogenesis of hepatocellular carcinoma, Liver International. 2008, 28 (2): 160-174.

Wong L.T., Yau T.O. and Ng I.O.L., The tumor suppressor phosphatase and tensin homolog (PTEN) in hepatocarcinogenesis, Proceedings of the Annual Meeting of American Association for Cancer Research, San Francisco, USA, April. 2008.

Wong Y.C., Yau T.O. and Ng I.O.L., Upregulation of the WNT co-receptor LRP6 in hepatocellular carcinoma, Proceedings of the Annual Meeting of American Association for Cancer Research, San Francisco, USA, April. 2008.

Yam J.W.P., Wong C.M. and Ng I.O.L., Deleted in liver cancer 1 (DLC1), In: Schwab M, Encyclopedia of Cancer. Springer, 2008, 2nd Ed..

Yam J.W.P., Tse Y.T., Ko C.F., Chan L.K., Sze M.F. and Ng I.O.L., Tensin2 with growth suppression and apoptosis induction activities is underexpressed in hepatocellular carcinoma, 2008 Proceedings of the American Association for Cancer Research, San Diego, CA, USA, April. 2008.

Researcher : Nicholls JM

Project Title:	ACE and ACE 2 expression in lungs of normal patients and patients with SARS
Investigator(s):	Nicholls JM, Peiris JSM, Cheung CY
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	09/2005

Abstract:

In December 2002 - June 2003 Southern China, including Hong Hong, was affected by a new coronavirus that resulted in a high mortality compared to previous coronaviral infections. The aetiological agent SARS-CoV was identified at Hong Kong University(1). In late 2003 two groups of researchers (2,3) identified a putative receptor for SARS-CoV,angiotensin converting enzyme 2 (ACE2 ) in cell lines and showed that the spike protein of SARS-CoV binds to the ACE2 receptor.Angiotensin converting enzyme (ACE) is a key enzyme in the well known rennin-angiotensin-aldosterone system (RAAS) - an endocrine cascade which is involved in blood pressure regulation. Angiotensionogen is cleaved by renin to produce a pro-hormone angiotensin I - a decapeptide. ACE removes the terminal 2 amino acids ( His-Leu) to produce an active octapeptide angiotensin 2 which is a potent vasoconstrictor and stimulator of cardiac growth (3). Whilst ACE was discovered over 50 years ago, it was only in 2000 that 2 independent groups (4,5). identified another enzyme ACE2, which though similar in many features to ACE functions by only removing a single amino acid ( Phe), converting angiotensin II to the less damaging Ang 1-7. ACE2 is therefore considered to have opposing functions to ACE(6)There is an increasing body of evidence supporting the role of an activated renin- angiotensin system (RAS) in acute lung injury/acute respiratory distress syndrome as increased ACE has been identified in the bronchoalveolar fluid of patients with ARDS together with a decrease in circulating ACE(7). Recent publications have shown that ACE2 does have an opposing function to ACE in lung injury, and that it does have a protective role - in one study mice that were given recombinant ACE2 were protected from acid-induced lung injury(8). The mechanism this appears to be a balance between the two angiotensin receptors in the lung - AT1aR and AT2R. Angiotensin II binds to both AT1aR and AT2R. Binding to AT1aR by Ang II will stimulate lung injury whilst binding toAT2R reduces lung injury. What ACE2 does is to reduce the levels of Ang II thereby reducing the binding of the AT1aR and thus reducing lung injury. The same researchers as publication (8) have also proposed that binding of SARS-CoV to ACE2 donwregulates this protective function of ACE2 by internalizing the receptor, thus allowing the angiotensin II to produce the lung damage(9).There is however some uncertainty about to what extent ACE and ACE2 are expressed in the respiratory tract. In the original reports detecting ACE2 in humans, Northern blots identified ACE2 in the heart, kidney and testis, but lungs were negative. Whilst Harmer and colleagues have recently used PCR to identify ACE2 in the gastrointestinal tract with a lower expression in the lungs(10) there seems to be a discrepancy between these findings and the widespread immunohistochemical detection of ACE2 in organ systems (11). Furthermore there is no data available concerning the ACE and ACE2 expression in the paediatric respiratory tract which would be of interest as SARS was not as fatal in this age group than in the elderly.The key issues therefore are as follows:1. Is there a correlation between the immunohistochemical detection of ACE/ACE 2 and PCR detection in the lungs?2. What cells of the respiratory tract contain ACE/ACE2?3. Is there a difference in expression of ACE and ACE2 in paediatric lung tissues and adult lung tissues?4. Do the lungs from patients who died of SARS show any change in ACE/ACE2 expression than normal patients? REFERENCES:1. Peiris JS, Lai ST, Poon LL, Guan Y, Yam LY, Lim W, Nicholls J, Yee WK, Yan WW, Cheung MT, Cheng VC, Chan KH, Tsang DN, Yung RW, Ng TK, Yuen KY; SARS study group Coronavirus as a possible cause of severe acute respiratory syndrome. Lancet. 2003;361:1319-252. Li W, Moore MJ, Vasilieva N, Sui J, Wong SK, Berne MA, Somasundaran M, Sullivan JL, Luzuriaga K, Greenough TC, Choe H, Farzan M. Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus.Nature. 2003;426:450-43. Xiao X, Chakraborti S, Dimitrov AS, Gramatikoff K, Dimitrov DS. The SARS-CoV S glycoprotein: expression and functional characterization. Biochem Biophys Res Commun. 2003 ;312:1159-647. Zisman LS. ACE and ACE2: a tale of two enzymes.Eur Heart J. 2005;26:322-44. Donoghue M, Hsieh F, Baronas E, Godbout K, Gosselin M, Stagliano N, Donovan M, Woolf B, Robison K, Jeyaseelan R, Breitbart RE, Acton S. A novel angiotensin-converting enzyme-related carboxypeptidase (ACE2) converts angiotensin I to angiotensin 1-9. Circ Res. 2000 ;87:E1-96. Tipnis SR, Hooper NM, Hyde R, Karran E, Christie G, Turner AJ. A human homolog of angiotensin-converting enzyme. Cloning and functional expression as a captopril-insensitive carboxypeptidase. J Biol Chem. 2000;275:33238-437. Turner AJ, Hiscox JA, Hooper NM. ACE2: from vasopeptidase to SARS virus receptor. Trends Pharmacol Sci. 2004 ;25:291-47. Idell S, Kueppers F, Lippmann M, Rosen H, Niederman M, Fein A. Angiotensin converting enzyme in bronchoalveolar lavage in ARDS. Chest. 1987;91:52-68.Imai Y, et al. Angiotensin-converting enzyme 2 protects from severe acute lung failure.Nature. 2005 ;436:112-69.Kuba K et al. A crucial role of angiotensin converting enzyme 2 (ACE2) in SARS coronavirus-induced lung injury.Nat Med. 2005 Jul 10; [Epub ahead of print]10. Harmer D, Gilbert M, Borman R, Clark KL. Quantitative mRNA expression profiling of ACE 2, a novel homologue of angiotensin converting enzyme. FEBS Lett. 2002;532:107-1011. Hamming I, Timens W, Bulthuis ML, Lely AT, Navis GJ, van Goor H. Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis. J Pathol. 2004 ;203:631-7

Project Title:	Sialic acid and receptor expression in normal respiratory tract, H5N1 influenza and non-avian influenza cases
Investigator(s):	Nicholls JM, Guan Y, Peiris JSM
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	10/2005

Abstract:

To determine if there is a change in expression of sialic acids in the conjunctiva and respiratory tract of nornal, avian influenza and fatal non-avian influenza cases that makes infection by the H5N1 more likely in Chinese subjects.

Project Title:	Occult respiratory viral infections in coronial autopsies - a pilot project
Investigator(s):	Nicholls JM, Peiris JSM, Chan KH, Beh SL, Poon LLM
Department:	Pathology
Source(s) of Funding:	Research Fund for the Control of Infectious Diseases - Full Grants
Start Date:	12/2005

Abstract:

Analyse the presence of clinically undiagnosed respiratory viral infections in autopies performed under HK coronial system

Project Title:	Respiratory Syncytial virus and Influenza associated cardiovascular mortality
Investigator(s):	Nicholls JM
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	04/2007

Abstract:

There is an increasing body of knowledge associating acute and chronic infections with cardiovascular diseases, principally ischaemic stroke and myocardial infarction (1) and also a seasonal influence on cardiovascular disease. This has initially been reported in geographical regions with cold winter climates, such as Japan (2) The United States of America (3) and Scotland (4) and this has been the rationale for vaccination against influenza (1,5) in order to reduce the mortality associated with myocardial infarction and stroke (6). The pathophysiological mechanisms associated with influenza and cardiovascular disease may be due to changes in clotting factors, cytokine concentrations and concentrations of inflammatory-response proteins that enhance thrombosis, impair vasodilation or lead to endothelial cell injury (7,8).It is possible however that some of these pro-thrombotic factors may be due to cold weather, rather than infection per se as haemoconcentration due to cold and changes in cardiovascular reflexes are attributed to cold temperatures (9).Whilst the previous reports have dealt with influenza disease burden in temperate regions, data on the tropical or sub-tropical regions is scanty. In 2004 we reported that Hong Kong had a similar influenza associated mortality compared to temperate regions (10) with influenza contributing to 17.1 to 21.5 excess deaths due to IHD per 100,000 population aged more than 65 years. In this study the effect of respiratory syncytial virus was not considered to be significant. As previous statistical models of morbidity and mortality due to influenza have been criticized on the grounds that RSV may contribute to mortality some studies have specifically used periods where RSV is not prevalent. To date the studies associating influenza with IHD have been based on statistical models and data provided by government census and statistics departments which though effective may be affected by ICD classification of death and mis-reporting. In addition studies indicate a discrepancy rate from 4% to 50% for clinical causes of death and those found at post-mortem (11). There have been no large scale studies which have looked at the association between viral respiratory disease and post-mortem findings. Hong Kong is in a favourable position from this respect because it has an active forensic pathology service which deals with all deaths outside public hospitals in Hong Kong. In the calendar year 2004, of the 7040 deaths due to natural causes, 3673 were due to diseases of the circulatory system and autopsies were performed in 1530 of these cases. (In 2143 cases the autopsy was waived because even though the death was reported to the coroner there was sufficient clinical and laboratory information to ascertain a cause of death) (12). Nevertheless, these 1530 cases represent a valuable source of material which has the potential to give a direct link between viral respiratory infection and cardiovascular disease and it is this material which will be used for this research.We have previously shown that it is possible to amplify viral RNA from archival tissues which together with immunohistochemical analysis enables a retrospective diagnosis of infection due to Influenza or RSV to be made. Since an accurate cause of cardiovascular death will be determined as an autopsy has been performed and the coronary arteries and myocardium have been examined, the possibility of misdiagnosis will be reduced.

List of Research Outputs

Chan K.H., Peiris J.S.M., Lim W., Nicholls J.M. and Chiu S.S.S., Comparison of nasopharyngeal flocked swabs and aspirates for rapid diagnosis of respiratory viruses in children , Journal of Clinical Virology. 2008, 42(1): 65-9.

Chan M.C.W., Chan W.Y., Yu C.L., Nicholls J.M., Chui W.H., Guan Y. and Peiris J.S.M., Polarity of influenza H5N1 virus infection in human respiratory epithelial cells, Health Research Symposium 2007.

Chan W.Y., Yu C.L., Nicholls J.M., Tsao G.S.W., Peiris J.S.M. and Chan M.C.W., Innate immune response and virus replication in human nasopharyngeal epithelial cells infected with influenza A (H5N1) virus, American Society for Microbiology general meeting 2008.

Chan W.Y., Yu C.L., Nicholls J.M., Chui W.H., Guan Y., Peiris J.S.M. and Chan M.C.W., The use of in vitro and ex vivo model to study the tropism and pathogenesis of emerging respiratory viruses, Health Research Symposium 2007.

Haselhorst T., Garcia J., Islam T., Lai J.C.C., Rose F.J., Nicholls J.M., Peiris J.S.M. and von Itzstein M., Avian influenza H5-containing virus-like particles (VLPs): host-cell receptor specificity by STD NMR spectroscopy, Angewandte Chemie (International ed. in English). 2008, 47(10): 1910-2.

Imai Y., Kuba K., Neely G.G., Yaghubian-Malhami R., Perkmann T., Van Loo G., Ermolaeva M., Veldhuizen R., Leung C.Y.H., Wang H., Liu H., Sun Y., Pasparakis M., Kopf M., Mech C., Bavari S., Peiris J.S.M., Slutsky A.S., Akira S., Hultqvist M., Holmdahl R., Nicholls J.M., Jian C., Binder C.J. and Penninger J.M., Identification of oxidative stress and Toll-like receptor 4 signaling as a key pathway of acute lung injury., Cell. Elsevier, 2008, 133(2): 235-249.

Lee M.Y., Cheung C.Y., Nicholls J.M. and Peiris J.S.M., Induction of COX-2 mediated proinflammatory cascade during H5N1 infection, Health Research Symposium 2007.

Leung G.K.K., Lee C.K. and Nicholls J.M., Radiation-induced rhabdomyosarcomatous transformation of a recurrent meningeal haemangiopericytoma. , Acta Neurochir (Wien). 2007, Epub ahead of print.

Nicholls J.M. and Peiris J.S.M., Avian influenza: update on pathogenesis and laboratory diagnosis, Respirology. 2008, 13(Suppl 1): S14-S18.

Nicholls J.M. and Garcia J., Determination of the haemagglutinin - sialic acid receptor interaction of H5N1 and other influenza viruses at the atomic level, Research on Infectious Diseases, Pasteur Insitute, Paris. 2008.

Nicholls J.M., Chan W.Y., Russell R.J., GM A.I.R. and Peiris J.S.M., Evolving complexities of influenza virus and its receptors, Trends in Microbiology. 2008, 16(4): 149-157.

Nicholls J.M. and Peiris J.S.M., Ex vivo tissues and emerging viral infections, International Consortium for Antivirals, Toronto, Canada. 2008.

Nicholls J.M., Chan M.C., Chan W.Y., Wong H.K., Cheung C.Y., Kwong D.L.W., Wong M.P., Chui W.H., Poon L.L., Tsao S.W., Guan Y. and Peiris J.S.M., Jropism of avian influenza A (H5N1) in the upper and lower respiratory tract, Nat Med. 2007, 13: 147-9.

Nicholls J.M., Bourne A.J., Chen H., Guan Y. and Peiris J.S.M., Sialic acid receptor detection in the human respiratory tract: evidence for widespread distribution of potential binding sites for human and avian influenza viruses, Respiratory Research. 2007, 8: 73.

Nicholls J.M. and Peiris J.S.M., The use of ex-vivo tissues for studying infection by emerging viruses, Bangkok International Conference on Avian Influenza, 23-25 January 2008.. 2008.

Nicholls J.M., Update on Avian Influena Pathogenesis, Annual meeting of Asian Pacific Society for Respirology, Gold Coast, Australia. . 2007.

Wu E.X., Wu Y., Nicholls J.M., Liao S., Lau C.P. and Tse H.F., MR diffusion tensor imaging study of postinfarct myocardium structural remodeling in a porcine model, In: Wu EX, Magn Reson Med. . 58(4), 2007, 687-95.

Zhao X., Nicholls J.M. and Chen Y.G., Severe acute respiratory syndrome-associated coronavirus nucleocapsid protein interacts with Smad3 and modulates transforming growth factor-beta signaling, The Journal of Biological Chemistry. 2008, 283(6): 3272-80.

Researcher : Shek TWH

List of Research Outputs

Au W.Y., Lo S.H., Law W.L., Khong P.L., Shek T.W.H., Lau W.H. and Chan E.M.C., Concomitant EBER+ve lymphomatoid papulosis and malignant transformation of colonic polyposis in a heart transplant recipient, Journal of Heart and Lung Transplantation. 2008, 27: 575-576.

Au W.Y., Leung A.Y.H., Tse E.W.C., Cheung W.W., Shek T.W.H. and Kwong Y.L., High incidence of tuberculosis after alemtuzumab treatment in Hong Kong Chinese patients, Leukemia Research. 2007, 32(4): 547-551.

Au W.Y., Liu C.L., Tam S., Fong B.M., Shek T.W.H., Hui C.K. and Kwong Y.L., Oral arsenic trioxide therapy for acute promyelocytic leukemia before and after liver transplantation for hepatitis B virus-related liver failure, Annals of Hematology. 2007, 86(10): 771-2.

Cheuk D.K.L., Shek T.W.H., Chan G.C.F., Lau Y.L., Ha S.Y. and Chiang A.K.S., Parotid acinic cell carcinoma in a long-term survivor of childhood acute lymphoblastic leukemia, Pediatr Blood Cancer. 2008 ;50(3):636-9. 2008, 50(3): 636-9.

Chim J.C.S., Yiu C.P.B. and Shek T.W.H., Pathological bone fracture in non-Hodgkin's lymphoma, Journal of Clinical Oncology. 2007, 25(21): 3175-6.

Chow K.C., Chan G.C.F., Khong P.L., Shek T.W.H., Ha S.Y. and Lau Y.L., Primary Ki-1-positive anaplastic large cell lymphoma of the bone presented with spinal cord compression, Hong Kong Journal of Paediatrics. 2007, 12: 283-286.

Hui C.K., Leung N., Shek T.W.H., Zhang H., Luk J.M.C., Poon R.T.P., Lo C.M., Fan S.T. and Lau G., Changes in liver histology as a surrogate endpoint of antiviral therapy for chronic HBV can predict progression to liver complications, Journal of Clinical Gastroenterology. 2008, 42(5): 533-538.

Hui C.K., Leung N., Shek T.W.H., Yao H., Lee W.K., Lai J.Y., Lai S.T., Wong W.M., Lai S.L., Poon R.T.P., Lo C.M., Fan S.T. and Lau G., Sustained disease remission after spontaneous HBeAg seroconversion is associated with reduction in fibrosis progression in chronic hepatitis B Chinese patients , Hepatology. 2007, 46(3): 690-698.

Hwang Y.Y., Chim J.C.S. and Shek T.W.H., Multiple myeloma with testicular involvement, Journal of Clinical Oncology: official Journal of the American Society of Clinical Oncology. 2008, 26(9): 1558-9.

Yuan X.J., Chan G.C.F., Chan S.K., Shek T.W.H., Kwong D.L.W., Wei W.I., Ha S.Y. and Chiang A.K.S., Treatment outcome of rhabdomyosarcoma in Hong Kong Chinese children, Hong Kong Medical Journal. 2008, 14(2): 116-123.

Researcher : Shen L

List of Research Outputs

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Shen L., Au W.Y., Guo T., Wong K.Y., Wong M.L.Y., Tsuchiyama J., Yuen P.W., Kwong Y.L., Liang R.H.S. and Srivastava G., Proteasome inhibitor bortezomib-induced apoptosis in natural killer (NK)-cell leukemia and lymphoma: an in vitro and in vivo preclinical evaluation, Blood. 2007, 110(1): 469-70.

Researcher : Siu KY

Project Title:	Ectoplasmic specialization dynamics in the testis is regulated by focal adhesion complex-mediated signaling pathways
Investigator(s):	Siu KY, Lee WWM
Department:	Zoology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2004

Abstract:

To investigate the downstream signaling pathway(s) underneath integrin and FAK that contribute to the regulation of ES dynamics.

Project Title:	p21-activated kinases (Paks) in ovarian and endometrial cancers
Investigator(s):	Siu KY, Cheung ANY
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	01/2006

Abstract:

The purpose of the proposed investigation is to characterize the expression levels and the signaling pathways of Paks in ovarian and endometrial cancers in an attempt to explore the roles of Paks in their carcinogenesis, with potential impacts on the development of targeted anticancer therapies. Key Issues and Problems being addressed: Ovarian and endometrial cancers are two common gynecological cancers. In 2002, ovarian cancer ranked eighth among the major causes of cancer death in Hong Kong. Endometrial cancer is the most common gynecological cancer worldwide. To have a greater chance to cure, it is important to define the staging of cancers at the first diagnosis. Recent studies in our laboratories have reported several tumor suppressor genes as potential cancer markers, which play a vital role in gynecological malignancies [1-5]. The high mortality rate of cancers is related to metastasis, which is a process involving alternation in signaling pathways, increased directional cell migration and enhanced cell survival [6-8]. There is accumulating evidence that actin cytoskeleton reorganization is an essential step for cell migration, anchourage-independent growth, and invasion [7]. Members of small GTPases of the Rho family (RhoA, Rac1, and Cdc42) have been found to be an important regulator for cytoskeleton reorganization [7]. RhoA is involved in stress fiber formation, Rac1 is for lamellipodia formation and Cdc42 is for filopodia formation [6, 9]. In mammalian cells, p21-activated serine/threonine kinases (Paks) were initially identified as an effector molecules of Rac1 and Cdc42. Consequently, they were found to be activated by both GTPase-dependent and -independent manners [6, 9]. To date, six members of Paks have been found. They can be classified into two subgroups: Group 1, consisting of Pak1 (Pak), Pak2 (Pak), and Pak3 (Pak), which share >90% homology in their kinase domain and 73% overall homology in their entire sequence; and Group 2, consisting of Pak4-6, which share only ~50% homology with the Pak1 kinase domain [6, 9]. For GTPase-dependent activation manner, Paks bind with Rac1 and Cdc42 in their active, GTP bound state, inhibiting their intrinsic GTPase activity and leading to Paks autophosphorylation. Once phosphorylated, they dissociate from Rac1/Cdc42 to phosphorylate downstream effectors. Although all Paks can bind to GTPases, only Group I Paks can be activated through this binding [6, 9]. In addition, Paks can be activated through a variety of GTPase-independent mechanisms, including direct phosphorylation by other kinases, such as protein kinase B (PKB, also know as Akt) under the influence of lipid signaling involving PI 3-kinase [6, 9, 10]. Due to their complicated activation processes and the presence of multiple downstream effectors, Paks play an important role in a wide range of cellular processes, including cytoskeletal reorganization, cell motility, apoptosis, cell cycle progression, and cell transformation, all of which are required for cancer progression [6, 10, 11]. The differential expression of Paks in ovarian and endometrial cancers is poorly defined. Only Pak1 has been found to be overexpressed in ovarian cancers [12]. Worst yet, the upstream mediators and the downstream signaling pathway(s) of Paks that leads to cancer progression in ovarian and endometrial cancers are still unknown. References (Publications from our group): 1.* Chan QKY, Khoo US, Ngan HYS, Yang CQ, Xue WC, Chan KYK, Chiu PM, Ip PPC, Cheung ANY. Single Nucleotide Polymorphism of Pi-class Glutathione-S-Transferase and Susceptibility to Endometrial Carcinoma. Clin. Cancer Res. 2005; 11: 2981-2985. 2.* Chan KYK, Ozcelik H, Cheung ANY, Ngan HYS, Khoo US. Epigenetic factors controlling the BRCA1 and BRCA2 genes in sporadic ovarian cancer. Cancer Research 2002; 63: 4151-4156. 3.* Shen DH, Chan KYK, Khoo US, Ngan HYS, Xue WC, Chiu PM, Ip PPC, Cheung ANY. Epigenetic and genetic alterations of p33ING1b in ovarian cancer. Carcinogenesis 2005; 26: 855-863. 4.* Chan QKY, Khoo US, Chan KYK, Ngan HYS, Li SS, Chiu PM, Man LS, Ip PPC, Xue WC, Cheung ANY. Promoter Methylation and Differential Expression of pi-class Glutathione-S-Transferase in Endometrial Carcinoma. Journal of Molecular Diagnostics 2005; 7: 8-16. 5. The Hong Kong Cancer Registry. Department of Health Annual Report 2001/2002. Hong Kong, Department of Health, Hospital Authority. 2000. 6. Bokoch GM. Biology of the p21-activated kinases. Annu. Rev. Biochem. 2003; 72: 743-781. 7. Hall A. Rho GTPases and the actin cytoskeleton. Science 1998; 279: 509-514. 8. Vadlamudi RK, Kumar R. P21-activated kinases in human cancer. Cancer Metastasis Rev. 2003; 22: 385-393. 9. Hofmann C, Shepelev M, Chernoff J. The genetics of Pak. J. Cell Sci. 2004; 117: 4343-4354. 10.* Siu MKY, Wong CH, Lee WM, Cheng CY. Sertoli-germ cell anchoring junction dynamics in the testis are regulated by an interplay of lipid and protein kinases. J. Biol. Chem. 2005; 280: 25029-25047. 11. Kumar R, Hung MC. Signaling intricacies take center stage in cancer cells. Cancer Res. 2005; 65: 2511-2515. 12. Schraml P, Schwerdtfeger G, Burkhalter F, Raggi A, Schmidt D, Ruffalo T, King W, Wilber K, Mihatsch MJ, Moch H. Combined array comparative genomic hybridization and tissue microarray analysis suggest PAK1 at 11q13.5-q14 as a critical oncogene target in ovarian carcinoma. Am. J. Pathol. 2003; 163: 985-992.

List of Research Outputs

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Over-expression of Prostatic Stem Cell Antigen (PSCA) is associated with Gestational Trophoblastic neoplasia. , Histopathology. 2007, 52: 167-174.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Overexpression of prostate stem cell antigen is associated with gestational trophoblastic neoplasia, Histopathology. 2008, 52(2): 167-174.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Liao X., Siu K.Y., Au W.H., Wong E.S.Y., Ngan H.Y.S. and Cheung A.N.Y., Aberrant Activation of Hedgehog Signaling Pathway in Ovarian Cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Siu K.Y., Chan H.Y., Kong S.H., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Differential expression of folate receptor alpha and reduced folate carrier and effect of folate in ovarian cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Siu K.Y., Woo N.W., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th Annual Growth Factor and Signal Transduction Symposium titled "Senescence, Aging and Cancer",Iowa, 26-29 July . 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Li A.S.M., Chan Y.K. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress 2007, 14-16 Nov. 2007.

Zhang H., Siu K.Y., Li A.S.M., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Wong K.Y., Li A.S.M., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Oct4 is Epigenetically Regulated by Methylation in Normal Placenta and Gestational Trophoblastic Disease, Placenta. 2008, 29(6): 549-554.

Researcher : So JCC

List of Research Outputs

Au W.Y., Pang A.W.K., Lam K.Y., Song Y., Lam W.M., So J.C.C. and Kwong Y.L., G6PD deficiency from lyonization after hematopoietic stem cell transplantation from female heterozygous donors, Bone Marrow Transplantation. 2007, 40(7): 677-681.

Au W.Y., Wong K.Y., Wan T.S.K., So J.C.C., Srivastava G. and Wong H.M., Very late relapse of B cell lymphoma masquerading as blastic transformation of chronic myeloid leukemia, Leuk Lymphoma. 2007, 48(7): 1414-1416.

Gibney G.T., Panhuysen C.I.M., So J.C.C., Ma E.S.K., Ha S.Y., Li C.K., Lee A.C.W., Li C.K., Yuen H.L., Lau Y.L., Johnson D.M., Farrell J.J., Bisbee A.B., Farrer L.A., Steinberg M.H., Chan L.C. and Chui D.H.K., Variation and heritability of Hb F and F-cells among beta-thalassemia heterozygotes in Hong Kong., Am J Hematol. 2008, 458-464.

So J.C.C., Wan T.S.K., Yip S.F. and Chan L.C., A dual colour dual fusion fluorescence in situ hybridisation study on the genesis of complex variant translocations in chronic myelogenous leukaemia, Oncol Rep. 2008, 19(5): 1181-1184.

So J.C.C., Ma E.S.K., Wan T.S.K., Yip S.F. and Chan L.C., Clinicopathological features of unbalanced translocation Der(1;7)(q10;p10) in myeloid neoplasms, Leuk Res. 2008, 32(6): 1000-1001.

So J.C.C., Diagnosis of globin gene deletions using multiplex ligation-dependent probe amplification, XXIst International Symposium on “Technological Innovations in Laboratory Hematology” of the International Society for Laboratory Hematology, Sydney, 1 May. 2008.

So J.C.C., Interpretation of thalassaemia/haemoglobinopathy investigations results, Pathology COC (MT/MLT) Commissioned Training Programme 2007-2008, Hong Kong, 16 Nov. 2007.

Researcher : So KT

List of Research Outputs

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Srivastava G

Project Title:	Role of CD44 and protein kinase C-binding protein 1, novel translocation partners of immunoglobulin heavy chain gene, in the pathogenesis of gastric lymphoma
Investigator(s):	Srivastava G
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	09/2004

Abstract:

To investigate the role of CD44 and PRKCBP1. For CD44 study: (1) to determine whether the gene amplification of CD44 represents another mechanism of CD44 upregulation in GL cases with strong CD44 expression. (2) to determine whether the hypermethylation of the promoter region of CD44 gene may be responsible for the downregulation of CD44 in GL cases lacking CD44 expression. (3) to determine whether the lack of CD44 mRNA expression detected in the pre- B-cell lines and some mature B-cell lines is due to hypermethylation of the promoter region of CD44 gene and whether the silencing of CD44 in the these cell lines could be partially reversed by the treatment of these cell lines with the demethylation agent 5-aza-2'-deoxycytidine (5-aza-^dC). (4) to detect the somatic mutations in CD44 cDNA and genomic DNA in GL, as the presence of mutations at hyaluronan binding sites of CD44 may affect its normal biological function. (5) to identify different CD44 splicing variants (CD44v) in GL since a correlation between expression of CD44v and adverse prognosis has been reported in several subtypes of human carcinoma and in NHL. (6) to correlate the findings of the IGH/CD44 translocation, gene amplification of CD44, hypermethylation of CD44 promoter, CD44 expression, somatic mutations in CD44 cDNA and the different CD44 splicing variants in GL with the clinico-pathological features (CPC) to define prognostic subgroups. (7) to clone the wild-type cDNA CD44 and CDNA lacking the leader peptide, as the result of JGH/CD44 translocation in GL, and perform the transfection assays for their cellular localization and to determine whether this variant of CD44 without the leader peptide has acquired additional tumor transforming function. For PRKCBPI study: (1)to examine PRKCBP1 mRNA expression in primary B cells by in-situ hybridization in normal lymphoid tissues including reactive tonsil, lymph node and spleen. (2) to examine PRKCBP1 mRNA expression by Northern blot hybridization and RT-PCR in the normal peripheral B-lymphocytes and in human B-lineage tumor cell lines corresponding to different stages of B-cell development. (3) to determine whether the PRKCBP1 mRNA is differentially expressed in the tumor cells of GL. (4) to detect the somatic mutations in PRKCBP1 CDNA sequences are prone to mutations coding (a)₈ microsatellite in PRKCBP1 mRNA. Repetitive DNA sequences are prone to mutations. (5) to correlate the findings of the IGH/PRKCBP1 translocation, differential PRKCBP1 mRNA expression and somatic mutations in PRKCBP1 cDNA in GL with the CPC to define prognostic subgroups. (6) to clone the wild-type PRKCBP1 cDNA lacking the first 11 non-coding exons, as the result of IGH/PRKCBP1 translocation in GL, and perform the transfection studies for the cellular localization of their protein to predict possible function of this gene.

Project Title:	Characterization of the role of immunoglobulin heavy and light chain [kappa] and [lambda] gene translocations in the pathogenesis of gastric lymphoma
Investigator(s):	Srivastava G, Liang RHS
Department:	Pathology
Source(s) of Funding:	Michael Kadoorie Cancer Genetics Research Fund
Start Date:	01/2005

Abstract:

To clone IGH-related translocations in GL at both IGHJ and 5' S[mu] region by LDI-PCR and those involving switch region sequences 3' to the S[mu], S[gamma]1-4, and S[alpha]1-2 regions by LDV-PCR; to clone IG[kappa]- and IG[lambda]-related translocations involving the joining region in GL by LDI-PCR; to determine the frequency of the known and novel IG (IGH, IG[kappa], IG[lambda])-related translocations identified by us in GL by direct PCR on genomic DNA; to study the expression of novel IG translocation partner genes in GL by immunostaining to determine whether the over expression of these genes in the tumor cells is associated with the IG translocation of these genes in GL; to determine the mRNA expression of novel IG translocation partner genes in the normal peripheral B-cells and human B-lineage tumor cell lines that correspond to different stages of B-cell development, loss of the normal control mechanisms regulating expression of these novel IG partner genes may contribute to malignant transformation in lymphomas; to analyze the mutations of incoming novel IG translocation partner genes in GL since in B cells, where the somatic hypermutation mechanism is active, mutations of the incoming gene may be observed and may contribute to the neoplastic phenotype; to determine the transforming and tumorigenic potential of each of the overexpressed novel IG translocation partner genes by in vitro growth properties and in vivo tumorigenicity assays; to determine the prevalence of all the cloned novel IG-related translocations identified by us in GL in nodal specimens (MZBCL and DLBCL) by direct PCR, for comparison of IG translocations in gastric MALT and DLBCL with their respective nodal counterparts.

Project Title:	Molecular mechanisms and clinicopathological significance of the aberrant nuclear BCL10 expression and constitutive NF-[kappa]B activation in EBV-associated nasal NK/T-cell lymphoma
Investigator(s):	Srivastava G, Liang RHS
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	10/2005

Abstract:

To analyze BCL10 and NF-χB expression in the cytoplasmic and nuclear protein fractions prepared from normal NK cells, NL specimens and NK tumor cell lines by Western blot analysis NF-χB transfer from cytoplasm to nucleus is the hallmark of its activation; to determine whether the nuclear localization of BCL10 in N1 is due to constitutive hyperphosphorylation of endogenous BCL10 by Western blot analysis and also by metabolic labeling experiments of NK tumor cell lines using ³²P-orthophosphate; to determine whether t(1;14)(p22; q32) or t(11;18)(q21;q21), which are associated with the BCL10 nuclear expression in gastric MALT lymphoma, are also present in NL; to determine whether BCL10 nuclear expression present in NL is due to other chromosomal translocations involving the BCL10 gene locus (1p22) by 5' RACE(5' rapid amplification of cDNA ends), resulting either (i) in the upregulation of BCL10 due to promoter substitution, or (ii) in fusion gene products involving BCL10 and novel fusion partner genes; for comparison, determine the subcellular localization of BCL10 and NF-χB expression by immunohistochemistry in tumor biopsy specimens of (i) nasal-type NK/T cell lymphoma occurring in non-nasal areas, (ii) aggressive NK cell lymphoma/leukemia, and (iii) non-nasal peripheral T-cell lymphoma (PTCL); and examine the status of EBV infection in these nasal-type NK/T-cell lymphoma lymphomas & non-nasal PTCL by EBV EBER in-situ hybridization on cases with nuclear BCL10, as it is possible that BCL10 nuclear localization is related to EBV infection; to determine the usefulness of the detection of aberrant nuclear BCL10 and NF-χB expression immunohistochemistry as a potential tumor marker for the diagnosis of NL (and also in nasal-type NK/T cell lymphomas & non-nasal PTCL with nuclear BCL10 expression as detected in objective#5); to examine whether nuclear expression of BCL10 expression or NFχB activation can be used as an independent prognostic marker to indicate more advanced or more aggressive NL (and also in nasal-type NK/T-cell lymphomas & non-nasal PTCL with nuclear BCL10 expression as detected in objective#5).

Project Title:	To identify tumor suppressor genes in the commonly deleted region of 6q in nasal NK/T-cell lymphoma by employing array-CGH and epigenetic approach
Investigator(s):	Srivastava G, Liang RHS, Tao Q
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	03/2006

Abstract:

Putative natural killer (NK) cell lymphoma/leukemia is a rare group of recently characterized hematolymphoid malignancies. They are highly aggressive and frequently present in extranodal sites, including the nasal area and the upper aerodigestive system, and non-nasal areas such as the skin and the gastrointestinal tract. By clinicopathological features, they can be classified into nasal NK/T-cell lymphoma (NL), nasal-type NK/T-cell lymphoma occurring in non-nasal areas, and NK cell lymphoma/leukemia. Primary NL represent the second most frequent group of extra-nodal lymphomas in Hong Kong Chinese. They are reported mostly from Asia, Mexico, and South America, but are extremely rare in Western countries. So far there have been limited data on the genetic aberrations in NL. However, preliminary results from a small number of NL cases investigated by karyotyping, metaphase-CGH and LOH studies have consistently shown a commonly deleted region of 6q21-25 in NL. Refinement of allelic loss at 6q might lead to the identification of novel TSG candidates involved in the tumorigenesis of NL. Unfortunately, owing to marked tumor necrosis and the difficulty of obtaining adequate biopsy materials from the nasal areas, procurement of the large amount of tumor tissue that are required for a comprehensive search for allelic loss at this region is not feasible. Moreover, this approach will require a laborious search for nearby genes by position cloning once the smallest altered locus is identified. The discovery of numerous hypermethylated promoters of TSG, along with a better understanding of gene-silencing mechanisms, has moved the subject of DNA methylation from obscurity to recognition of methylation as an important alternative mechanism of TSG inactivation in cancer. The incorporation of epigenetic analysis methods into microarray technology can yield quantitative and qualitative information on DNA methylation throughout the genome. This epigenetic approach to identify TSG candidates in NL is now feasible for NL due to the availability of authentic NL and aggressive NK-cell lymphoma/leukemia cell lines. The overall aim of this study is to identify TSG candidates in the commonly deleted region of 6q in NL first by array-CGH of NL cell lines and, second by employing an epigenetic approach based on subtractive, cancer genomic and microarray analyses. Clone-based array-CGH, with an average resolution of 1.3 Mb, have been previously applied successfully to gain insights into the pathogenetic mechanisms of cancer development.Major Aims:1. To perform array-CGH to map deletions in NL and aggressive NK-cell lymphoma/leukemia cell lines for further refinement of allelic loss in the commonly deleted region of chromosome 6q in NL. 2. To identify TSG candidates in NL by the detection of upregulated transcripts after pharmacological demethylation and HDAC inhibition of TSG promoters in NL cell lines by: (1) cDNA microarray hybridization using GeneChip® expression arrays from Affymetrix® using the cDNA subtracted between mock-treated and DAC- and TSA-treated 5 NL and 4 aggressive NK-cell lymphoma/leukemia cell lines as the probe, and (2) by direct PCR screening on the subtracted cDNA for gene transcripts from the chromosome region 6q25.2-q25.3 and other specific regions deleted at 6q as identified by array-CGH in this study. 3. To determine the status of DNA hypermethylation by methylation-specific PCR (MSP) and bisulfite genomic sequencing (BGS) of the TSG promoters in 6q (identified in this study by the combination of 1 and 2 above) and correlate the methylation status of these gene promoters with the down-regulation of the expression of these genes detected by semi-quantitative RT-PCR in the NL and aggressive NK-cell lymphoma/leukemia cell lines and NL tumor biopsy specimens.4. To perform the tumor suppressive functional assays of the selected TSG candidates in 6q (identified in this study by the combination of 1, 2 and 3) by standard transfection and colony formation assays.

Project Title:	Therapeutic potential and mechanisms of antitumor activity of proteasome inhibitor bortezomib in extranodal NK/T-cell lymphoma, nasal type
Investigator(s):	Srivastava G, Liang RHS
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	09/2006

Abstract:

To examine whether bortezomib promotes apoptosis and inhibits cell growth in vitro in a large series of NK lymphoma/leukemia cell lines and extranodal NK/T-cell lymphoma, nasal type (NL) primary cultures by analyzing cell viability, cell senescence, cell cycle and apoptosis; (2) to investigate whether bortezomib is effective in treating NL in vivo, we will study the effect of bortezomib on the treatment of NL NUDE mouse model HX-NKTL. In addition, two NL cell lines will be implanted subcutaneously in nude mice to provide tumor xenograft animal models, including HANKI1 that is a xenograft-derived NL cell line. The drug will be injected intravenously to determine its effectiveness; (3) bortezomib directly affects two important intracellular signaling pathways-NF-κB and JNK that are shown to be regulated by ubiquitin-proteasome degradation in multiple myeloma cells. We will investigate whether this anticancer drug works by the similar molecular mechanisms in NL; (4) to further understand the mechanisms of antitumor action of bortezomib, we will examine the molecules associated with the G₂/M checkpoint because we detected G₂/M arrest in a drug-treated NL cell line in a pilot study; (5) because the drug induced apoptosis in most of the analyzed cell lines and mitochondria are belived to play a crucial role in the control of apoptosis, apoptotic signaling molecules, especially those located in mitochondria, will be analyzed to uncover their involvement with the action of bortezomib; we will explore the essential molecular changes in the early stages of drug action. Superoxide production and intracellular Ca²⁺ dysregulation may be the critical determinants, so we will identify their roles in the bortezomib-induced apoptosis pathways.

List of Research Outputs

Au W.Y., Wong K.Y., Wan T.S.K., So J.C.C., Srivastava G. and Wong H.M., Very late relapse of B cell lymphoma masquerading as blastic transformation of chronic myeloid leukemia, Leuk Lymphoma. 2007, 48(7): 1414-1416.

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Chan S.L., Cui Y., van Hasselt A., Li H., Srivastava G., Jin H., Ng K.M., Wang Y., Lee K.Y., Tsao G.S.W., Zhong S., Robertson K.D., Rha S.Y., Chan A.T. and Tao Q., The tumor suppressor Wnt inhibitory factor 1 is frequently methylated in nasopharyngeal and esophageal carcinomas, Lab Invest. 2007, 87(7): 644-50.

Cheung C.M., Tang J.C.O., Lee P.Y., Hu L., Guan X.Y., Tang W.K., Srivastava G., Wong J., Luk J.M.C. and Law S.Y.K., Establishment and characterization of a new xenograft-derived human esophageal squamous cell carcinoma cell line HKESC-4 of Chinese origin, Cancer Genetics and Cytogenetics. 2007, 178(1): 17-25.

Chim J.C.S., Wong K.Y., Loong F., Lam W.W. and Srivastava G., Frequent epigenetic inactivation of Rb1 in addition to p15 and p16 in mantle cell and follicular lymphoma, Hum Pathol. 2007, 38(12): 1849-57.

Chim J.C.S., Wong K.Y., Loong F., Lam W.W. and Srivastava G., Frequent epigenetic inactivation of Rb1 in addition to p15 and p16 in mantle cell and follicular lymphoma, Human Pathology. 2007, 38(12): 1849-57.

Fu L., Qin Y.R., Xie D., Hu L., Kwong D.L.W., Srivastava G., Tsao G.S.W. and Guan X.Y., Characterization of a novel tumor-suppressor gene PLC delta 1 at 3p22 in esophageal squamous cell carcinoma, Cancer Res. 2007, 67(22): 10720-6.

Jin H., Wang X., Ying J., Wong A.H., Cui Y., Srivastava G., Shen Z.Y., Li E.M., Zhang Q., Jin J., Kupzig S., Chan A.T., Cullen P.J. and Tao Q., Epigenetic silencing of a Ca2+-regulated Ras GTPase-activating protein RASAL defines a new mechanism of Ras activation in human cancers, Proc Natl Acad Sci U S A. 2007, 104(30): 12353-12358.

Ko J.M.Y., Chan P.L., Yau W.L., Chan H.K., Chan K.C., Yu Z.Y., Kwong F.M., Miller L.D., Liu E.T., Yang L.C., Lo P.H.Y., Stanbridge E.J., Tang J.C.O., Srivastava G., Tsao G.S.W., Law S.Y.K. and Lung M.L., Monochromosome transfer and microarray analysis identify a critical tumor-suppressive region mapping to chromosome 13q14 and THSD1 in esophageal carcinoma, Molecular Cancer Research. 2008, 6(4): 592-603.

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Shen L., Au W.Y., Guo T., Wong K.Y., Wong M.L.Y., Tsuchiyama J., Yuen P.W., Kwong Y.L., Liang R.H.S. and Srivastava G., Proteasome inhibitor bortezomib-induced apoptosis in natural killer (NK)-cell leukemia and lymphoma: an in vitro and in vivo preclinical evaluation, Blood. 2007, 110(1): 469-70.

Srivastava G., Editor of International Journal of Medical Sciences. Ivyspring International Publisher, 2007.

Ying J., Li H., Chen W.Y.W., Srivastava G., Gao Z. and Tao Q., WNT5A is epigenetically silenced in hematologic malignancies and inhibits leukemia cell growth as a tumor suppressor, Blood. 2007, 110(12): 4130-1.

Yuen H.F., Chan Y.P., Chan K.K., Chu Y.Y., Wong M.L.Y., Law S.Y.K., Srivastava G., Wong Y.C., Wang X. and Chan K.W., Id-1 and Id-2 are markers for metastasis and prognosis in oesophageal squamous cell carcinoma, Br J Cancer. 2007, 97(10): 1409-15.

Researcher : Sun Q

List of Research Outputs

Sun Q., Kong C.T., Huang F. and Chan L.C., Aberrant Dendritic Cell Differentiation Initiated by Mll-Een Fusion Gene Does Not Requre Leukaemic Transformation., Journal of Leukocyte Biology. 2008, 83 (1): 173 - 180.

Researcher : Sze MF

List of Research Outputs

Sze M.F. and Ng I.O.L., Application of an inducible gene expression system to functionally study hepatitis B virus X protein, Health Research Symposium 2007: Building bridges between research, practice & policy, September 29. 2007.

Yam J.W.P., Tse Y.T., Ko C.F., Chan L.K., Sze M.F. and Ng I.O.L., Tensin2 with growth suppression and apoptosis induction activities is underexpressed in hepatocellular carcinoma, 2008 Proceedings of the American Association for Cancer Research, San Diego, CA, USA, April. 2008.

Researcher : Szeto EF

List of Research Outputs

Cheung A.N.Y., Tsun O.K.L., Szeto E.F., Wong G., Lo S. and Ngan H.Y.S., Application of Novel Markers to Cervial Cytology for enhancing Cervical Cancer Screening, Annual Scientific Meeting of Hong Kong Society of Cytology, Hong Kong, 8-9 December. 2007.

Cheung A.N.Y., Tsun O.K.L. and Szeto E.F., ThinPrep Imaging System in Cervical Cytology Screening, Annual Scientific Meeting of Hong Kong Society of Cytology, Hong Kong, 8-9 December. 2007.

Researcher : Tam S

List of Research Outputs

Au W.Y., Lam W.M., Chu W.C., Tam S., Wong W.K., Pennel D.J., Lie A.K.W. and Liang R.H.S., A Magnetic Resonance Imaging Study of Iron Overload in Hemopoietic Stem Cell Transplant Recipients With Increased Ferritin Levels, Transplantation Proceedings. 2007, 39(10): 3369-3374.

Au W.Y., Lam W.W., Chu W.W., Tam S., Wong W.K., Liang R.H.S. and Ha S.Y., A T2* magnetic resonance imaging study of pancreatic iron overload in thalassemia major, Haematologica. 2008, 93(1): 116-119.

Au W.Y., Lam W.W.M., Chu W.W.C., Tam S., Wong W.K., Chan H., Law M.F., Liu H.S.Y. and Liang R.H.S., A pilot MRI study of organ specific hemosiderosis and functional correlation in Chinese patients with myelodysplasia and aplastic anemia with raised ferritin levels, Hematological Oncology. E Pub, 2008.

Au W.Y., Tam S. and Kwong Y.L., Entry of elemental arsenic into the central nervous system in patients with acute promyelocytic leukemia during arsenic trioxide treatment , Leukemia Research. 2007, 32(2): 357-358.

Au W.Y., Liu C.L., Tam S., Fong B.M., Shek T.W.H., Hui C.K. and Kwong Y.L., Oral arsenic trioxide therapy for acute promyelocytic leukemia before and after liver transplantation for hepatitis B virus-related liver failure, Annals of Hematology. 2007, 86(10): 771-2.

Au W.Y., Tam S., Fong B.M., Ho K.L., Tam P.C. and Kwong Y.L., Prolonged oral arsenic trioxide therapy and neprholithiasis, Leukemia and Lymphoma . 2007, 48(11): 2233-2234.

Au W.Y., Tam S., Fong B.M., Wan T.S.K., Yip S.F. and Kwong Y.L., Second hematological malignancies during arsenic trioxide therapy of B-cell lymphomas , Leukemia Research. EPub, 2008.

Fung C.W., Blau N., Siu S., Mak C.M., Cheung P.T., Tam S. and Wong V.C.N., A new presentation of 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency Dopa Responsive Dystonia (DRD), 12th National Child Neurology Congress (第十二届全國小兒神經學術會議), Wuyishan, China, 27-29 October, 2007.

Fung C.W., Siu S., Mak C.M., Poon G.W.K., Wong K.Y., Cheung P.T., Low L.C.K., Tam S. and Wong V.C.N., A rare cause of hepatosplenomegaly transaldolase deficiency, 12th National Child Neurology Congress (第十二届全國小兒神經學術會議), Wuyishan, China, 27-29 October, 2007.

Fung C.W., Siu S., Mak C.M., Poon G.W.K., Wong K.Y., Cheung P.T., Low L.C.K., Tam S. and Wong V.C.N., A rare cause of hepatosplenomegaly transaldolase deficiency, A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Cheung P.T., Tam S. and Wong V.C.N., A rare presentation of 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency - Dopa Responsive Dystronia (DRD), A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Cheung P.T., Mak C.M., Siu S., Tam S. and Wong V.C.N., A study of cerebrospinal fluid neurotransmitters assay in children with undiagnosed neurological diseases in Hong Kong, Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society and HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Tam S. and Wong V.C.N., An atypical presentation of cerebral folate deficiency, 12th National Child Neurology Congress (第十二届全國小兒神經學術會議), Wuyishan, China, 27-29 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Tam S. and Wong V.C.N., An atypical presentation of cerebral folate deficiency, A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Fung C.W., Blau N., Siu S., Mak C.M., Tam S. and Wong V.C.N., An atypical presentation of cerebral folate deficiency, Annual Symposium of Society for the Study of Inborn Errors of Metabolism, Hamburg, 4-7 September 2007.

Lam W.W., Au W.Y., Chu W.C., Tam S., Ha S.Y. and Pennell D.J., One-stop measurement of iron deposition in the anterior pituitary, liver, and heart in thalassemia patients, Journal of Magnetic Resonance Imaging. 2008, 28(1): 29-33.

Lui S.L., Tsang R.C.W., Chan K.W., Zhang Q., Tam S., Yung S.S.Y. and Chan D.T.M., Rapamycin attenuates the severity of established nephritis in lupus-prone NZB/W F1 mice, Nephrology Dialysis Transplantation. 2008, 23: 2768-2776.

Lui S.L., Yung S.S.Y., Tsang R.C.W., Zhang Q., Chan K.W., Tam S. and Chan D.T.M., Rapamycin prevents the development of nephritis in lupus-prone NZB/W F1 mice, Lupus. 2008, 17: 305-313.

Luk H.M., Fung C.W., Mak C.M., Lam C.W., Tam S. and Wong V.C.N., Severe myoclonic epilepsy in infancy (SMEI) or Dravet Syndrome - case report with SCN1A mutation in Hong Kong, A Joint Annual Scientific Meeting for Paediatricians and Paediatric Nurses (organized by The HK Paediatric Society & HK Paediatric Nurses Association), Hong Kong, 27 October, 2007.

Tse K.C., Yung S.S.Y., Tang S.C.W., Tam S., Lai K.N. and Chan D.T.M., Atorvastatin at conventional dose did not reduce C-reactive protein in patients on peritoneal dialysis, Journal of Nephrology. 2008, 21: 283.

Wong K.F., Tam S. and Kwong Y.L., Diagnosis of familial amyloidotic polyneuropathy by bone marrow biopsy, British Journal of Haematology. 2007, 139(4): 517.

Wong K.Y., Wong R.M.S., Wong M.S.C. and Tam S., Reference Values for Thyroid Function Tests in Asian Very Low Birth Weight Infants, The 4^th Congress of Asian Society for Pediatric Research (ASPR), Honolulu, Hawaii, USA, 3-6 May 2008. 122.

Yeung C.M., Tan Y., Tam S., Lu L., Ko K.H., Yang P., Kung H.F. and Lin M.C., The Antidiabetic Effects of a Dry Powder of Dietary Vegetable and Fruit Mixtures in Diabetic db/db Mice, Biologics: Targets and Therapy. 2008, 2(3): 571-576.

Researcher : Tam WY

List of Research Outputs

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Researcher : Tam YSI

List of Research Outputs

Leung L.H., Tam Y.S.I., Tin P.C., Chung L.P. and Wong M.P., Src Kinase Promotes Survival and Invasion of Non-Small Cell Lung Cancer cells with Epidermal Growth Factor Receptor Abnormalities and is a Potential Candidate for Combination Targeted Therapy , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008, San Diego, California, the United State . 2008.

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Tang JCO

List of Research Outputs

Cheung C.M., Tang J.C.O., Lee P.Y., Hu L., Guan X.Y., Tang W.K., Srivastava G., Wong J., Luk J.M.C. and Law S.Y.K., Establishment and characterization of a new xenograft-derived human esophageal squamous cell carcinoma cell line HKESC-4 of Chinese origin, Cancer Genetics and Cytogenetics. 2007, 178(1): 17-25.

Ko J.M.Y., Chan P.L., Yau W.L., Chan H.K., Chan K.C., Yu Z.Y., Kwong F.M., Miller L.D., Liu E.T., Yang L.C., Lo P.H.Y., Stanbridge E.J., Tang J.C.O., Srivastava G., Tsao G.S.W., Law S.Y.K. and Lung M.L., Monochromosome transfer and microarray analysis identify a critical tumor-suppressive region mapping to chromosome 13q14 and THSD1 in esophageal carcinoma, Molecular Cancer Research. 2008, 6(4): 592-603.

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Researcher : Tao Q

List of Research Outputs

Chan S.L., Cui Y., van Hasselt A., Li H., Srivastava G., Jin H., Ng K.M., Wang Y., Lee K.Y., Tsao G.S.W., Zhong S., Robertson K.D., Rha S.Y., Chan A.T. and Tao Q., The tumor suppressor Wnt inhibitory factor 1 is frequently methylated in nasopharyngeal and esophageal carcinomas, Lab Invest. 2007, 87(7): 644-50.

Jin H., Wang X., Ying J., Wong A.H., Cui Y., Srivastava G., Shen Z.Y., Li E.M., Zhang Q., Jin J., Kupzig S., Chan A.T., Cullen P.J. and Tao Q., Epigenetic silencing of a Ca2+-regulated Ras GTPase-activating protein RASAL defines a new mechanism of Ras activation in human cancers, Proc Natl Acad Sci U S A. 2007, 104(30): 12353-12358.

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Ying J., Li H., Chen W.Y.W., Srivastava G., Gao Z. and Tao Q., WNT5A is epigenetically silenced in hematologic malignancies and inhibits leukemia cell growth as a tumor suppressor, Blood. 2007, 110(12): 4130-1.

Researcher : Tin PC

List of Research Outputs

Leung L.H., Tam Y.S.I., Tin P.C., Chung L.P. and Wong M.P., Src Kinase Promotes Survival and Invasion of Non-Small Cell Lung Cancer cells with Epidermal Growth Factor Receptor Abnormalities and is a Potential Candidate for Combination Targeted Therapy , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008, San Diego, California, the United State . 2008.

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Trendell-Smith NJ

List of Research Outputs

Au W.Y., Yeung C.K., Cheung M.C. and Trendell-Smith N.J., Penile lichen sclerosus after allogeneic stem cell transplantation, British Journal of Dermatology. EPub, 2008.

Researcher : Tse YT

List of Research Outputs

Yam J.W.P., Tse Y.T., Ko C.F., Chan L.K., Sze M.F. and Ng I.O.L., Tensin2 with growth suppression and apoptosis induction activities is underexpressed in hepatocellular carcinoma, 2008 Proceedings of the American Association for Cancer Research, San Diego, CA, USA, April. 2008.

Researcher : Tsui WY

List of Research Outputs

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Kosinski C., Li V.S.W., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors, Proc Natl Acad Sci U S A. 2007, 104(39): 15418-23.

Li V.S.W., Kosinski C., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression profiling of human colon top versus basal crypts and identification of BMP antagonists as candidates for intestinal stem cell niche, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Researcher : Tsun OKL

List of Research Outputs

Cheung A.N.Y., Tsun O.K.L., Szeto E.F., Wong G., Lo S. and Ngan H.Y.S., Application of Novel Markers to Cervial Cytology for enhancing Cervical Cancer Screening, Annual Scientific Meeting of Hong Kong Society of Cytology, Hong Kong, 8-9 December. 2007.

Cheung A.N.Y., Tsun O.K.L. and Szeto E.F., ThinPrep Imaging System in Cervical Cytology Screening, Annual Scientific Meeting of Hong Kong Society of Cytology, Hong Kong, 8-9 December. 2007.

Researcher : Wan TSK

List of Research Outputs

Au W.Y., Tam S., Fong B.M., Wan T.S.K., Yip S.F. and Kwong Y.L., Second hematological malignancies during arsenic trioxide therapy of B-cell lymphomas , Leukemia Research. EPub, 2008.

Au W.Y., Wong K.Y., Wan T.S.K., So J.C.C., Srivastava G. and Wong H.M., Very late relapse of B cell lymphoma masquerading as blastic transformation of chronic myeloid leukemia, Leuk Lymphoma. 2007, 48(7): 1414-1416.

So J.C.C., Wan T.S.K., Yip S.F. and Chan L.C., A dual colour dual fusion fluorescence in situ hybridisation study on the genesis of complex variant translocations in chronic myelogenous leukaemia, Oncol Rep. 2008, 19(5): 1181-1184.

So J.C.C., Ma E.S.K., Wan T.S.K., Yip S.F. and Chan L.C., Clinicopathological features of unbalanced translocation Der(1;7)(q10;p10) in myeloid neoplasms, Leuk Res. 2008, 32(6): 1000-1001.

Researcher : Wang X

Project Title:	Downregulation of MAD2 expression and its significance in chemodrug sensitization in nasopharyngeal carcinoma cells
Investigator(s):	Wang X, Nicholls JM, Tsao GSW, Jin D
Department:	Anatomy
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	09/2003

Abstract:

To confirm that MAD2 expression is downregulated in clinical samples of NPC; to determine the mechanism by which MAD2 is downregulated in NPC; to establish whether upregulation of MAD2 expression can sensitize NPC cells to chemotherapy.

Project Title:	The role fo MAD2 in overcoming cisplatin resistance in testicular germ cell tumors
Investigator(s):	Wang X, Jin D
Department:	Anatomy
Source(s) of Funding:	Lance Armstrong Foundation - General Award
Start Date:	10/2003

Abstract:

To investigate the association between decreased MAD2 protein expression and cisplatin resistance in TGCT cells; to study if exogenous expression of the MAD2 gene in TGCT cells can lead to chemosensitization to cisplatin in TGCT cells; to demonstrate that downregulation of MAD2 results in resistance ot cisplatin in TGCT cells; to characterize the role of MAD2 in cisplatin-induced cell death inTGCT cells; to identify binding partners of MAD2 in response to cisplatin-induced DNA damage.

Project Title:	Significance of MAD2 expression to chromosomal instability in prostate cancer
Investigator(s):	Wang X, Jin D Y
Department:	Anatomy
Source(s) of Funding:	Association for International Cancer Research - General Award
Start Date:	04/2004

Abstract:

To correlate MAD2 expression with genomic instability in prostate cancer specimens; to show that MAD2 expression is essential for a functional mitotic checkpoint in prostate cancer cells; to demonstrate that downregulation of MAD2 leads to mitotic checkpoint defect and increased CIN in prostate cancer cells. • To investigate if promoter hypermethylation contributes to decreased MAD2 expression in prostate cancer

Project Title:	Molecular mechanisms involved in the suppressive effects of garlic derivatives on cell growth and motility in prostate cancer cells
Investigator(s):	Wang X, Jin D Y
Department:	Anatomy
Source(s) of Funding:	American Institute for Cancer Research (AICR) - General Award
Start Date:	08/2005

Abstract:

To study the role of the mitotic checkpoint in SAC and SAMC-induced androgen independent prostate cancer cell death; to investigate if SAC and SAMC have any inhibitory effect on the invasive ability of prostate cancer cells and to determine the molecular mechanisms responsible. Specific Aim 3. To examine if SAC and SAMC can enhance the sensitivity of prostate cancer cells to chemotherapeutic drugs.

Project Title:	Significance of Id-1 upregulation and its association with EGFR in bladder cancer
Investigator(s):	Wang X
Department:	Anatomy
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	06/2006

Abstract:

Background: Bladder cancer is one of the common urological cancers. Although majority of bladder cancers are superficial at presentation, 20% of the bladder cancer patients present with muscle invasive disease at diagnosis (i.e. stage T2-T4 tumours). If untreated, fewer than 15% of those patients can survive more than two years. Two main challenges remain for the treatment of this cancer which are (i) the ability to identify the small but significant number of patients with non-muscle-invasive disease who will progress to muscle-invasive disease, and (ii) to improve current treatment of the muscle invasive bladder cancer. Recently, our group identified a potential oncogene, Id-1 (inhibitor of differentiation or DNA binding), and demonstrated its significance in the development of human prostate cancer (Ouyang et al., J Urol 167: 2598, 2002), nasopharyngeal carcinoma (Wang et al., 35:42-49, 2002) and ovarian cancer (Zhang et al., Br J Cancer 91: 2042, 2004). Since then, upregulation of Id-1 has been reported in over 20 types of human cancer such as breast, pancreas, cervical, melanoma, and head and neck (Reviewed by Wong et al., 9:279-289, 2004). In addition, increased Id-1 expression levels are associated with advanced tumour stage as well as poor prognosis (Maruyama et al., Am J Pathol 155: 815, 1999; Schindl et al., Cancer Res 61: 5703, 2001; Ouyang et al., J Urol 167: 2598, 2002). Furthermore, patients with higher levels of Id-1 have much shorter overall survival than the patients with relatively lower Id-1 expression in ovarian cancer (Schindl et al., Clin Cancer Res 9:779, 2003). Recent evidence also shows that Id-1 is able to induce epidermal growth factor receptor (EGFR) expression, indicating that the oncogenic effect of Id-1 may be mediated through activation of the EGFR pathway (Ling et al., Carcinogenesis 25: 517, 2004; Zhang et al., Br J Cancer 91: 2042, 2004). In bladder cancer, upregulation of EGFR has been frequently reported in tissue samples (Neal et al., Lancet 1: 366, 1985; Mellon et al., J Urol 153: 919, 1995). Several studies have correlated EGFR positively with tumour stage, tumour progression, and poor clinical outcome in bladder cancer patients (Neal et al., Cancer 65: 1619, 1990; Mellon et al., J Urol 153: 919, 1995; Nguyen et al., Am J Clin Pathol 101: 166, 1994). Furthermore, it was recently reported that treatment of bladder cancer cells with ZD1839 (or Iressa), a highly selective EGFR inhibitor, resulted in radiosensitization to ionizing radiation (Maddineni et al., Br J Cancer 92: 125, 2005), indicating that EGFR may be a potential therapeutic target in improving treatment efficiency of bladder cancer. Based on the evidence that Id-1 is able to activate EGFR pathway, we hypothesized that activation of the EGFR pathway observed in bladder cancer may be a result of overexpression of Id-1, as indicated in other cancers (Ling et al., Carcinogenesis 25: 517, 2004; Zhang et al., Br J Cancer 91: 2042, 2004 ). Purpose of the project: The aim of this project is to study the significance of Id-1 in bladder cancer and provide an alternative target of the EGFR pathway for the treatment of bladder cancer. This will be achieved through the following objectives: 1). To study differential Id-1 protein expression between normal and malignant bladder cancer specimens and correlate its expression levels with EGFR in different stage bladder cancer specimens. 2). To study if ectopic Id-1 expression in bladder cancer cell lines could lead to upregulation of EGFR and demonstrate the significance of Id-1 overexpression in bladder cancer cell proliferation. 3). To study if inactivation of Id-1 could suppress EGFR expression and investigate if downregulation of Id-1 could lead to inhibition of bladder cancer cell growth.

Researcher : Wong CM

Project Title:	Genetic and epigenetic alterations of serine protease inhibitor TFPI-2 in hepatocellular carcinoma.
Investigator(s):	Wong CM, Ng IOL
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	10/2005

Abstract:

Liver cancer is a major malignancy in the world and is particularly prevalent in this region, being the second most common fatal cancer in Asia including Hong Kong. Delineation of alterations or dysregulations of major cellular pathways in liver cancer is important as this can provide novel opportunities for diagnosis, clinical significance, prognosis and therapeutic interventions for patients with this cancer. As part of our long-standing effort in search for and identification of tumor suppressor genes important in liver cancer development and progression, we have used high-throughput microarray approach to screen for methylation in gene silencing in liver cancer (hepatocellular carcinoma, HCC) and have successfully selected a number of putative tumor suppressor genes (Wong et al., 2004). One of the candidate genes we identified with this technology is TFPI-2 (tissue-factor pathway inhibitor-2), which is a newly identified Kunitz-type serine protease inhibitor. TFPI-2 is abundantly expressed in umbilical vein endothelial cells, placenta, and liver. TFPI-2 inhibits gelatinolysis and caseinolysis by trypsin, plasmin, α-chymotrypsin, and pancreatic elastase (Rao et al 1995). TFPI-2 is believed to have important roles in invasive growth of tumor cells. Pervious, studies have been indicated that TFPI-2 involved in inhibiting degradation of extracellular matrix (ECM) in fibrosarcoma cell model (Rao et al., 1998). Our preliminary data have shown that this gene is frequently underexpressed and epigenetically silenced by methylation in human HCC cells and human HCC samples. In this proposal, we aim to delineate its roles in HCC and aim to 1) characterize the expression of TFPI-2 in human liver cancers and evaluate their clinicopathological and prognostic significance, 2) delineate the epigenetic and genetic alterations of TFPI-2 in liver cancer, and 3) assess the biochemical properties and anti-tumorigenic functions of TFPI-2 in liver cancer. Findings derived from this study may be of potential use for the development of effective and novel therapeutic interventions to target these genes in liver cancer. Reference Rao CN, Gomez DE, Woodley DT, Thorgeirsson UP. Novel extracellular matrix-associated serine proteinase inhibitors from human skin fibroblasts. Arch Biochem Biophys 1995; 317: 311-4. Rao CN, Cook B, Liu Y, Chilukuri K, Stack MS, Foster DC, Kisiel W, Woodley DT. HT-1080 fibrosarcoma cell matrix degradation and invasion are inhibited by the matrix-associated serine protease inhibitor TFPI-2/33 kDa MSPI. Int J Cancer 1998 ; 76: 749-56

List of Research Outputs

Wong C.M. and Ng I.O.L., Frequent overexpression of SUV39H1 in human hepatocellular carcinoma, American Association for Cancer Research Special Meeting: Cancer Epigenetics, Boston, MA, USA, May. 2008.

Wong C.M. and Ng I.O.L., Molecular pathogenesis of hepatocellular carcinoma, Liver International. 2008, 28 (2): 160-174.

Yam J.W.P., Wong C.M. and Ng I.O.L., Deleted in liver cancer 1 (DLC1), In: Schwab M, Encyclopedia of Cancer. Springer, 2008, 2nd Ed..

Researcher : Wong CW

List of Research Outputs

Suehiro Y., Wong C.W., Chirieac L.R., Kondo Y., Shen L., Webb C.R., Chan Y.W., Chan A.S.Y., Chan T.L., Wu T.T., Rashid A., Hamanaka Y., Hinoda Y., Shannon R.L., Morris J., Issa J.P., Yuen S.T., Leung S.Y. and Hamilton S.R., Epigenetic-Genetic Interactions in the APC/WNT, RAS/RAF, and P53 Pathways in Colorectal Carcinoma, Clin Cancer Res. 2008, 14(9): 2560-9.

Researcher : Wong ESY

List of Research Outputs

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Over-expression of Prostatic Stem Cell Antigen (PSCA) is associated with Gestational Trophoblastic neoplasia. , Histopathology. 2007, 52: 167-174.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Overexpression of prostate stem cell antigen is associated with gestational trophoblastic neoplasia, Histopathology. 2008, 52(2): 167-174.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Li A.S.M., Siu K.Y., Wong E.S.Y., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Effect of demethylation and histone deacetylase inhibitors on the expression of stem cell related genes in choriocarcinoma cell lines, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Liao X., Siu K.Y., Au W.H., Wong E.S.Y., Ngan H.Y.S. and Cheung A.N.Y., Aberrant Activation of Hedgehog Signaling Pathway in Ovarian Cancer, 99th American Association for Cancer Research 2008 Annual Meeting, San Diego, 12-16 April. 2008.

Siu K.Y., Woo N.W., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th Annual Growth Factor and Signal Transduction Symposium titled "Senescence, Aging and Cancer",Iowa, 26-29 July . 2007.

Woo N.W.S., Wong E.S.Y., Chan H.Y., Chan K.Y.Q., Ngan H.Y.S., Tsao G.S.W. and Cheung A.N.Y., p21-activated kinase 4 in Ovarian Cancer: Its expression, localization and possible functional role, 16th annual Growth Factor and Signal Transduction Symposium, Iowa, 26-29 July. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Li A.S.M., Chan Y.K. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress 2007, 14-16 Nov. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Wong K.Y., Li A.S.M., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Oct4 is Epigenetically Regulated by Methylation in Normal Placenta and Gestational Trophoblastic Disease, Placenta. 2008, 29(6): 549-554.

Researcher : Wong KY

List of Research Outputs

Au W.Y., Wong K.Y., Wan T.S.K., So J.C.C., Srivastava G. and Wong H.M., Very late relapse of B cell lymphoma masquerading as blastic transformation of chronic myeloid leukemia, Leuk Lymphoma. 2007, 48(7): 1414-1416.

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Chim J.C.S., Wong K.Y., Loong F., Lam W.W. and Srivastava G., Frequent epigenetic inactivation of Rb1 in addition to p15 and p16 in mantle cell and follicular lymphoma, Hum Pathol. 2007, 38(12): 1849-57.

Chim J.C.S., Wong K.Y., Loong F., Lam W.W. and Srivastava G., Frequent epigenetic inactivation of Rb1 in addition to p15 and p16 in mantle cell and follicular lymphoma, Human Pathology. 2007, 38(12): 1849-57.

Law F.B.F., Chen W.Y.W., Wong K.Y., Ying J., Tao Q., Langford C., Lee P.Y., Law S.Y.K., Cheung R.W.L., Chui C.H., Tsao G.S.W., Lam A.K.Y., Wong J., Srivastava G. and Tang J.C.O., Identification of a novel tumor transforming gene GAEC1 at 7q22 which encodes a nuclear protein and is frequently amplified and overexpressed in esophageal squamous cell carcinoma, Oncogene. 2007, 26(40): 5877-5888.

Shen L., Au W.Y., Guo T., Wong K.Y., Wong M.L.Y., Tsuchiyama J., Yuen P.W., Kwong Y.L., Liang R.H.S. and Srivastava G., Proteasome inhibitor bortezomib-induced apoptosis in natural killer (NK)-cell leukemia and lymphoma: an in vitro and in vivo preclinical evaluation, Blood. 2007, 110(1): 469-70.

Researcher : Wong LT

List of Research Outputs

Wong L.T., Yau T.O. and Ng I.O.L., The tumor suppressor phosphatase and tensin homolog (PTEN) in hepatocarcinogenesis, Proceedings of the Annual Meeting of American Association for Cancer Research, San Francisco, USA, April. 2008.

Researcher : Wong MLY

List of Research Outputs

Chan K.K., Shen L., Guo T., Wong M.L.Y., Wong K.Y., Au W.Y., Lu L., Kwong Y.L., Liang R.H.S. and Srivastava G., IL2 induced NF-kB activation in nasal NK/T-cell lymphoma is mediated through Akt and BCL10, Keystone Symposia on Molecular and Cellular Biology: Lymphocyte Activation and Signaling, February 3-8, 2008, Snowbird Resort, Snowbird, Utah. 2008.

Shen L., Au W.Y., Guo T., Wong K.Y., Wong M.L.Y., Tsuchiyama J., Yuen P.W., Kwong Y.L., Liang R.H.S. and Srivastava G., Proteasome inhibitor bortezomib-induced apoptosis in natural killer (NK)-cell leukemia and lymphoma: an in vitro and in vivo preclinical evaluation, Blood. 2007, 110(1): 469-70.

Yuen H.F., Chan Y.P., Chan K.K., Chu Y.Y., Wong M.L.Y., Law S.Y.K., Srivastava G., Wong Y.C., Wang X. and Chan K.W., Id-1 and Id-2 are markers for metastasis and prognosis in oesophageal squamous cell carcinoma, Br J Cancer. 2007, 97(10): 1409-15.

Researcher : Wong MP

Project Title:	Identification of candidate cancer genes in regions of frequent chromosomal aberration in non-small cell lung cancers
Investigator(s):	Wong MP, Chung LP, Lam WK
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	09/2003

Abstract:

To define critical regions of imbalance in frequently altered chromosomal loci identified in local samples of non-small cell lung cancers (NSCLC) by microstatellite analysis; to identify target genes in the analyzed regions by comparison with genes listed in databases of the Human Genome Resources(NCBI); to study the structure and expression of the targeted genes in NSCLC of smokers and non-smokers, so as to verify their involvement and to look for genetic or epigenetic alterations.

Project Title:	Differentially expressed genes in lung cancer
Investigator(s):	Wong MP, Chung LP, Lam WK
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2005

Abstract:

To verify whether molecular targets identified through expression profile microarray screening procesures truly show aberrant expressions in extended samples of lung cancers; to evaluate whether the genes encoding these verified molecular targets show alterations or mutations, so that their primary roles in carcinogenesis are supported at the genomic level.

Project Title:	Mutational Analysis of Epidermal Growth Factor Receptor in Non-Small Cell Lung Cancer
Investigator(s):	Wong MP, Chua DTT
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2005

Abstract:

A. Purpose of study: To evaluate the incidence and patterns of EGFR mutation in NSCLC, and to correlate the findings with clinicopathological characteristics as well as clinical responses of the patients to gefitinib therapy. B. Problem being addressed and Key Issues: • Non-small cell lung cancer (NSCLC) is a common cancer in Hong Kong for both men and women. While >90% of lung cancers in men are related to tobacco,

<30% of those in women are associated with cigarette smoking, but the underlying carcinogenic mechanism is still unclear. • Patients with NSCLC show poor overall survival due to late presentation defying complete surgical resection. Responses to current protocols of chemotherapy also demonstrate limited success. • Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that plays a crucial role in cell proliferation, survival and differentiation. In recent years, various forms of therapeutic agents targeting EGFR have been developed, including monoclonal antibodies or inhibitors of tyrosine kinase (TKI). Gefitinib (Iressa) is an orally administrated TKI that has been tried for NSCLC treatment. • It has recently been observed that patients with NSCLC containing EGFR mutations show better clinical responses to gefitinib, particularly women who have never smoked, or whose lung cancers show bronchioloalveolar features. However, a statistically significant benefit in survival has not been consistently demonstrated in published clinical series. The reported findings cannot be extrapolated to apply to our population, as the profile of mutation may be different and the response correlation may be population-dependent. It is thus important to evaluate the pattern of EGFR mutations and response to gefitinib in our patients with NSCLC.>

Project Title:	Significance of Circulating Tumour Cells and Serum Nucleic Acid Markers in Non-small Cell Lung Cancer
Investigator(s):	Wong MP, Chua DTT, Ho JCM
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	11/2006

Abstract:

Purpose of Study To investigate the usefulness of circulating tumour cells, and serum EGFR mutant and expression level as tumour markers for evaluating chemotherapy treatment options and predicting tumour outcome in patients with non-small cell lung cancer. Key issues  In Hong Kong, the incidence and mortality rates of lung cancer are high for both men and women. It is the commonest cancer in men and, as in Western populations, tobacco smoking is the main cause. It is the 2nd commonest cancer in women. However, most female lung cancer patients are life-long non-smokers and, passive smoking, air pollution, cooking habits and genetic influences have been suggested as etiological factors (1). The overall prognosis of lung cancer depends on the tumour stage, with relatively good outcome of >50% 5 year survival for stage I patients, and poor outcome with <15% 5 year survival for late stage patients.  Tumour markers embrace different forms of indicators that can be detected in the peripheral circulation which are postulated to reflect the systemic load of tumour cells within the body. Measurement of the tumour markers at different stages of tumour progression may provide useful information on the status of the tumour, e.g. residual or recurrent disease. It is hypothesized that these may correlate with the patient outcome and prognosis.  Different treatment options have recently become available for patients with lung cancer. For late stage (IIIB and IV) tumours, palliative tumour reduction by chemotherapy and/or radiotherapy is used. New molecular targeted therapy using tyrosine kinase inhibitors (TKI) that antagonize epidermal growth factor receptor (EGFR) signalling has led to promising results in patients whose tumours harbour activating EGFR mutations or increased EGFR expression. EGFR mutations have been found in >70% of female non-smokers and about 20% of male smokers in our local population (2). The high frequencies of these abnormalities implicate that EGFR mutants or expression level could be used as potential markers to detect tumour cells in the peripheral blood for monitoring systemic spread of lung cancer.  In view of the poor patient prognosis, and because of the diversity of treatment options available, it is important that clinicians are able to predict the likely outcome and monitor the treatment response of patients with lung cancers. Currently, apart from the tumour stage, few, if any, definitive prognostic indicators are known.  Two forms of tumour markers have been investigated in recent years, namely, circulating tumour cells (CTC) and serum nucleic acids of specific molecules. CTC are tumour cells that have become detached from the parent tumour and entered into the peripheral circulation. They are present in variable quantities in different patients but by using the appropriate technology, their numbers can be quantified. In experiments targeted to investigate the relation between the actual spiked and detected number of tumour cells in controlled samples, a linear and significant correlation between the two was shown for a range from 5 to about 1200 tumour cells in the samples. In clinical studies, the numbers of CTC have been shown to be a useful indicator of survival in patients with metastatic breast cancer (3). Very few studies of CTC in lung cancers have been reported. Gauthier et al, who used electromagnetic separation to isolate tumour cells and TRAP-assay to measure their telomerase activity, found CTC in 11/15 (73%) of stage IIIB and IV non-small cell lung cancer patients compared to 0/30 healthy controls (4). Kraeft et al detected fluorescent anti-epithelial antibody-labeled CTC in 18/40 (45%) of centrifuged blood from small cell lung cancer patients (5). However, the relation of CTC to tumour stage, patient survival and response to treatment has not been reported.  Circulating serum RNA of various molecules has been analyzed in lung cancer patients, such as hnRNP-B1, CEA, Her2, TTF1, etc. Fleischhacker et al were able to detect circulating RNA for hnRNP-B1 and/or Her2 in all 18 lung cancer patients and none of 12 control subjects studied (5). Analyses of serum DNA for various abnormalities, such as aberrant methylation of various genes, RAS mutations, microsatellite instabilities, etc, have been used as a means of detecting systemic tumour load. The use of mutant EGFR or EGFR expression level as tumour markers for lung cancer monitoring has not been reported. Problems being addressed: 1. To identify and quantify the number of circulating tumour cells (CTC) in the peripheral blood of lung cancer patients during different stages of tumour progression, aiming to evaluate whether the amount of CTC correlates with disease recurrence, metastasis or patient prognosis. 2. To evaluate serum nucleic acids for activating mutations in the EGFR gene and EGFR expression levels, and correlate these with treatment response to TKI, as well as tumour progress and patient outcome.

List of Research Outputs

Leung L.H., Tam Y.S.I., Tin P.C., Chung L.P. and Wong M.P., Src Kinase Promotes Survival and Invasion of Non-Small Cell Lung Cancer cells with Epidermal Growth Factor Receptor Abnormalities and is a Potential Candidate for Combination Targeted Therapy , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008, San Diego, California, the United State . 2008.

Nicholls J.M., Chan M.C., Chan W.Y., Wong H.K., Cheung C.Y., Kwong D.L.W., Wong M.P., Chui W.H., Poon L.L., Tsao S.W., Guan Y. and Peiris J.S.M., Jropism of avian influenza A (H5N1) in the upper and lower respiratory tract, Nat Med. 2007, 13: 147-9.

Wong M.K., Wong M.P., Lam D.C., Sihoe A.D.L., Cheng L.C., Lam B., Ip M.S.M., Nakajima T., Yasufuku K., Lam W.K. and Ho J.C.M., Endobronchial ultrasound for diagnosis of synchronous primary lung cancers, Lung Cancer. 2008.

Wong M.P., Interstitial Lung Disease: How can a Pathologist help, Autumn Respiratory Seminar. Hong Kong Thoracic Society and the American College of Chest Physicians (HK & Macau Chapter). 2007.

Wong M.P., Interstitial Lung Diseases, Postgraduate Diploma in Diagnosis and Therapeutics in Internal Medicine. 2007.

Wong M.P., Molecular Genetics of Non-small Cell Lung Cancer Hong Kong Experience, Nevada Cancer Institute, Las Vagas, USA. 2008.

Wong M.P., Pathological Diagnosis of Recurrences – Difficulties and Solutions, Head and Neck Course. Dept of Surgery, HKU. 2008.

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Wong OH

List of Research Outputs

Chen Y.X., Man K., Ling G.S., Chen Y., Sun B., Cheng Q., Wong O.H., Lo C.K., Ng I.O.L., Chan L.C., Lau G., Lin S.C.L., Huang F. and Huang F.P., A crucial role for dendritic cell (DC) IL-10 in inhibiting successful DC-based immunotherapy: superior antitumor immunity against hepatocellular carcinoma evoked by DC devoid of IL-10, Journal of Immunology. 2007, 179(9): 6009-6015.

Researcher : Wong WS

List of Research Outputs

Wong W.S., So K.T., Leung L.H., Tin P.C., Tam Y.S.I., Chung L.P. and Wong M.P., EML4-ALK is a new oncogene in non-small cell lung carcinoma showing wild-type EGFR and K-RAS from non-smokers , American Association for Cancer Research Annual Meeting, 2008 April 12-16, 2008,San Diego, California, the United State . 2008.

Researcher : Wong YC

List of Research Outputs

Wong Y.C., Yau T.O. and Ng I.O.L., Upregulation of the WNT co-receptor LRP6 in hepatocellular carcinoma, Proceedings of the Annual Meeting of American Association for Cancer Research, San Francisco, USA, April. 2008.

Researcher : Xu MS

List of Research Outputs

Chan Y.K., Ching C.Y.J., Xu M.S., Cheung A.N.Y., Yip S.P., Lam L.Y.C., Lai S.T., Chu C.M., Wong A.T.Y., Song Y., Huang F., Liu W., Chung P.H., Leung G.M., Chow E.Y.D., Chan E.Y.T., Chan J.C.K., Ngan H.Y.S., Tam P.K.H., Chan L.C., Sham P.C., Chan V.S.F., Peiris J.S.M., Lin S.C.L. and Khoo U.S., Association of ICAM3 Genetic Variant with Severe Acute Respiratory Syndrome, The Journal of Infectious Diseases. 2007, 196: 271-280.

Researcher : Xue W

List of Research Outputs

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Over-expression of Prostatic Stem Cell Antigen (PSCA) is associated with Gestational Trophoblastic neoplasia. , Histopathology. 2007, 52: 167-174.

Feng H., Tsao G.S.W., Ngan H.Y.S., Xue W., Kwan H.S., Siu K.Y., Liao X., Wong E.S.Y. and Cheung A.N.Y., Overexpression of prostate stem cell antigen is associated with gestational trophoblastic neoplasia, Histopathology. 2008, 52(2): 167-174.

Researcher : Yam JWP

Project Title:	Characterization of tensin2, the binding partner of DLC1 tumor suppressor in liver cancer
Investigator(s):	Yam JWP, Ng IOL, Jin D
Department:	Pathology
Source(s) of Funding:	Competitive Earmarked Research Grants (CERG)
Start Date:	01/2007

Abstract:

To verify the DLC1-tensin2 interaction; to determine the impact of DLC1-tensin2 interaction on their biochemical properties; to delineate the biological functions of tensin2 and its implications in HCC; to evaluate the clinicopathological significance of tensin2 in human HCC.

Project Title:	Nucleocytoplasmic shuttling mechanism of DLC1 tumor suppressor in liver cancer
Investigator(s):	Yam JWP, Ng IOL
Department:	Pathology
Source(s) of Funding:	Seed Funding Programme for Basic Research
Start Date:	01/2007

Abstract:

Purpose of the proposed investigation The purpose of this study is to elucidate how the subcellular localization of DLC1 (Deleted in Liver Cancer 1) determines its cellular functions. The spatial distribution of a protein is tightly regulated and correlated to proper functioning of a protein. Here we hypothesize that DLC1 shuttles between cytoplasm and nucleus, which in turn regulates its tumor suppressive functions. Objectives 1) To elucidate the nucleocytoplasmic shuttling mechanism of DLC1 2) To delineate the distinctive functional roles of DLC1 in both cytoplasm and nucleus 3) To evaluate the clinical significance of the cytoplasmic and nuclear localization of DLC1 in liver cancer Key issues and problems being addressed Hepatocellular carcinoma (HCC) is a major malignancy worldwide and the second commonest fatal cancer in Asia and Hong Kong. However, the molecular mechanisms underlying the development and progression of HCC are poorly understood. DLC1, a RhoGAP (Rho GTPase activating protein) was identified as a tumor suppressor in HCC. Tumor suppressive role of DLC1 and its epigenetic regulation have been clearly demonstrated. Our previous work has shown that DLC1 is a tumor suppressor gene critically involved in hepatocarcinogenesis. DLC1 is located on chromosome 8p which is frequently deleted in human HCC (Chan et al., 2002). We have also detected homozygous deletion and underexpression of DLC1 in HCC cell lines and human samples (Ng et al., 2000). Hypermethylation may account for the transcriptional silencing of DLC1 gene expression (Wong et al., 2003). Functionally, we are the first group demonstrating the growth inhibitory effect of DLC1 in HCC (Ng et al., 2000). Its inhibitory role in cell migration and invasion of HCC cell lines have been demonstrated (Wong, Yam et al., 2005). Inhibition of GTPase activity of RhoA and Cdc42 in vitro by DLC1 was first demonstrated by us (Wong et al., 2003). In our recent finding, tensin2 is identified as the first and novel interacting partner of DLC1 (Yam et al., 2006). Ectopic expression of DLC1 and tensin2 showed that the interacting complex colocalized with vinculin at focal adhesions. However, sequence analysis of DLC1 revealed the presence of nuclear localization signal (NLS) and nuclear export sequences (NES) and these raise the question how the subcellular distribution of DLC1 is functionally regulated. Also, it was intriguing to observe that DLC1, thought to have primarily roles in cytosol, exhibited nuclear localization and therefore nucleocytoplasmic shuttling. Given the importance of DLC1 as a tumor suppressor, much effort has been focused on the genetic, epigenetic and functional analyses of DLC1. However, the fundamental question about the subcellular localization of DLC1 has never been clearly defined. Thus, precise examination of the subcellular localization of DLC1 and functional implications is warranted.

List of Research Outputs

Leung T.H.Y., Yam J.W.P. and Ng I.O.L., STARD13 (StAR-related lipid transfer (START) domain containing 13). , Atlas Genet Cytogenet Oncol Haematol.. 2007.

Yam J.W.P., Wong C.M. and Ng I.O.L., Deleted in liver cancer 1 (DLC1), In: Schwab M, Encyclopedia of Cancer. Springer, 2008, 2nd Ed..

Yam J.W.P., Focal adhesion localization, tensin binding activity and tumor suppressive functions of DLC1 tumor suppressor, Cancer Centres for Research Joint Retreat, The University of Hong Kong and Hong Kong University of Science and Technology. 2008.

Yam J.W.P., Molecular Basis of Tumor Suppressor Genes and Oncogenes in Human Cancers, Hong Kong International Cancer Congress . 2007.

Yam J.W.P., Molecular characterization of human hepatocellular carcinoma: Tumor suppressor genes, American Association for Cancer Research Annual Meeting. 2008.

Yam J.W.P., Tse Y.T., Ko C.F., Chan L.K., Sze M.F. and Ng I.O.L., Tensin2 with growth suppression and apoptosis induction activities is underexpressed in hepatocellular carcinoma, 2008 Proceedings of the American Association for Cancer Research, San Diego, CA, USA, April. 2008.

Researcher : Yau TO

Project Title:	The role of Frizzled-7, receptor of the WNT/β-catenin signalling pathway, in hepatocellular carcinoma: Implications in tumorigenesis and metastasis
Investigator(s):	Yau TO, Ng IOL
Department:	Pathology
Source(s) of Funding:	Small Project Funding
Start Date:	10/2006

Abstract:

Hepatocellular carcinoma (HCC) is second commonest in Asia and in Hong Kong. Hepatocarcinogenesis is closely associated with activation of the WNT/β-catenin signaling pathway (HCC) (Wong et al., 2001; Miyoshi et al., 1998; de La Coste A et al., 1998). In the canonical WNT/β-catenin signaling pathway, WNT ligands bind to the transmembrane receptors Frizzled (FZD) and low density lipoprotein receptor related protein (LRP), and the signal is transduced to cytoplasmic protein FRAT1 and Dishevelled (DVL) respectively. FRAT1 and DVL inhibit the APC complex, and β-catenin therefore accumulates. Subsequently, β-catenin enters the nucleus, forms complex with members of T-cell factor/lymphoid enhancing factor (TCF/LEF) and upregulates TCF/LEF-dependent transcription of target proto-oncogenes such as c-MYC (He et al. 1998) and cyclin D1 (Shtutman et al., 1999; Tetsu et al., 1999), and eventually promotes tumourigenesis. We and others have previously demonstrated overexpression of β-catenin, in membranous, cytoplasmic and/or nuclear compartments, in 41 (68%) of 60 HCCs (Wong et al., 2001). We also shown that downregulation of negative regulator HDPR1 may be one of the mechanisms leading to the accumulation of β-catenin in HCC (Yau et al, 2005). To further delineate the deregulation of this important pathway in HCC, we plan to investigate the role of WNT receptor molecules in this deregulation. One of these receptor molecules, FZD7, was recently demonstrated to be overexpressed in HCC (Merle et al., 2004 and 2005) and implicated to promote motility of HCC cells. However, the role of FZD7 in hepatocarcinogenesis and metastasis has not been fully characterized. In this research proposal, we propose to investigate the roles of FZD7 in hepatocarcinogenesis and metastasis. Objectives of this research proposal: 1.To study the expression of FZD7 at mRNA and protein levels in human HCCs and in HCC cell lines. 2.To investigate whether there are any mutations of FZD7 that will result in activation of the pathway. 3.To investigate whether upregulation of FZD7 in HCC is due to gene amplification. 4.To study the role of FZD7 in hepatocarcinogenesis in HCC cell lines. 5.To study the role of FZD7 in metastasis using orthotopic xenograft implantation in immunodeficient mice.

List of Research Outputs

Mak G.W.Y., Leong V.Y.L., Yau T.O., Ng I.O.L. and Ching Y.P., Functional characterization of C53 and its possible roles in tumorigenesis, Oncogenes and Human Cancer- The next 25 years / Nature-Centro Nacional de Investigaciones Oncologicas (CNIO) Conference . 2007.

Wong L.T., Yau T.O. and Ng I.O.L., The tumor suppressor phosphatase and tensin homolog (PTEN) in hepatocarcinogenesis, Proceedings of the Annual Meeting of American Association for Cancer Research, San Francisco, USA, April. 2008.

Wong Y.C., Yau T.O. and Ng I.O.L., Upregulation of the WNT co-receptor LRP6 in hepatocellular carcinoma, Proceedings of the Annual Meeting of American Association for Cancer Research, San Francisco, USA, April. 2008.

Researcher : Yeung CW

List of Research Outputs

Ip P.P.C., Lam K.W., Cheung C.L., Yeung C.W., Pun T.C., Chan K.Y.Q. and Cheung A.N.Y., Tranexamic Acid-associated Necrosis and Intralesional Thrombosis of Uterine Leiomyomas: A Clinicopathologic Study of 147 Cases Emphasizing the Importance of Drug-induced Necrosis and Early Infarcts in Leiomyomas , American Journal of Surgical Pathology. 2007, 31(8): 1215-1224.

Researcher : Yip SF

List of Research Outputs

Au W.Y., Tam S., Fong B.M., Wan T.S.K., Yip S.F. and Kwong Y.L., Second hematological malignancies during arsenic trioxide therapy of B-cell lymphomas , Leukemia Research. EPub, 2008.

So J.C.C., Wan T.S.K., Yip S.F. and Chan L.C., A dual colour dual fusion fluorescence in situ hybridisation study on the genesis of complex variant translocations in chronic myelogenous leukaemia, Oncol Rep. 2008, 19(5): 1181-1184.

So J.C.C., Ma E.S.K., Wan T.S.K., Yip S.F. and Chan L.C., Clinicopathological features of unbalanced translocation Der(1;7)(q10;p10) in myeloid neoplasms, Leuk Res. 2008, 32(6): 1000-1001.

Researcher : Yip SP

List of Research Outputs

Khoo U.S., Chan Y.K., Chan V.S.F., Ching C.Y.J., Yam L., Chu C.M., Lai S.T., Wong T.Y., Tam P.K.H., Yip S.P., Leung G.M., Lin C.L. and Peiris J.S.M., Role of polymorphisms of the inflammatory response genes and DC-SIGNR in genetic susceptibility to SARS and other infections, Hong Kong Med J. 2008, 14(4)(Suppl 4): S9-12.

Researcher : Yuen HF

List of Research Outputs

Howard E.W., Leung C.L., Yuen H.F., Chua C.W., Lee D.T.W., Chan K.W., Wang X. and Wong Y.C., Decreased adhesiveness, resistance to anoikis and suppression of GRP94 are integral to the survival of circulating tumor cells in prostate cancer, Clinical and Experimental Metastasis. 2008, 25: 497-508.

Kwok W.K., Ling M.T., Yuen H.F., Wong Y.C. and Wang X., Role of p14^ARF in TWIST-mediated senescence in prostate epithelial cells, Carcinogenesis. 2007, 28(12): 2467-2475.

Yuen H.F., Chan Y.P., Chan K.K., Chu Y.Y., Wong M.L.Y., Law S.Y.K., Srivastava G., Wong Y.C., Wang X. and Chan K.W., Id-1 and Id-2 are markers for metastasis and prognosis in oesophageal squamous cell carcinoma, Br J Cancer. 2007, 97(10): 1409-15.

Researcher : Yuen HF

List of Research Outputs

Kwok W.K., Ling M.T., Yuen H.F., Wong Y.C. and Wang X., Role of p14^ARF in TWIST-mediated senescence in prostate epithelial cells, Carcinogenesis. 2007, 28(12): 2467-2475.

Researcher : Yuen ST

List of Research Outputs

Chan T.L., Tsui W.Y., Chan Y.W., Chan A.S.Y., Lee T.Y.H., Yuen S.T. and Leung S.Y., Germline epimutation of MLH1 in early-onset colorectal cancer, 99th Annual Meeting of the American Association for Cancer Research, April 12-16, 2008, San Diego, C.A., U.S.A.. 2008.

Chan T.L., Yuen S.T., Kong C.K., Chan Y.W., Chan A.S.Y., Ng W.F., Tsui W.Y., Lo M.W.S., Tam W.Y., Li V.S.W. and Leung S.Y., Research Output Prize "Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer, Nature Genetics. 2006, 38(10): 1178-83", The University of Hong Kong. 2007.

Kosinski C., Li V.S.W., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors, Proc Natl Acad Sci U S A. 2007, 104(39): 15418-23.

Li V.S.W., Kosinski C., Chan A.S.Y., Zhang J., Ho C., Tsui W.Y., Chan T.L., Mifflin R.C., Powell D.W., Yuen S.T., Leung S.Y. and Chen X., Gene expression profiling of human colon top versus basal crypts and identification of BMP antagonists as candidates for intestinal stem cell niche, Keystone Symposium on “Frontiers in Gastrointestinal Cancer: Molecular Genetics, Inflammation, Early Detection and Therapy”, Beijing, China, 14-19 October. 2007.

Schetter A.J., Leung S.Y., Sohn J.J., Zanetti K.A., Bowman E.D., Yanaihara N., Yuen S.T., Chan T.L., Kwong D.L., Au G.K., Liu C.G., Calin G.A., Croce C.M. and Harris C.C., MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma, JAMA. 2008, 299(4): 425-36.

Suehiro Y., Wong C.W., Chirieac L.R., Kondo Y., Shen L., Webb C.R., Chan Y.W., Chan A.S.Y., Chan T.L., Wu T.T., Rashid A., Hamanaka Y., Hinoda Y., Shannon R.L., Morris J., Issa J.P., Yuen S.T., Leung S.Y. and Hamilton S.R., Epigenetic-Genetic Interactions in the APC/WNT, RAS/RAF, and P53 Pathways in Colorectal Carcinoma, Clin Cancer Res. 2008, 14(9): 2560-9.

Sääf A.M., Halbleib J.M., Chen X., Yuen S.T., Leung S.Y., Nelson W.J. and Brown P.O., Parallels between Global Transcriptional Programs of Polarizing Caco-2 Intestinal Epithelial Cells In Vitro and Gene Expression Programs in Normal Colon and Colon Cancer, Mol Biol Cell. 2007, 18(11): 4245-60.

Researcher : Zhang H

List of Research Outputs

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, 14-16 Nov. 2007.

Chan H.Y., Siu K.Y., Wong E.S.Y., Zhang H., Ngan H.Y.S. and Cheung A.N.Y., Expression of phospho-Stat3 in hydatidiform mole, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Li A.S.M., Chan Y.K. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress 2007, 14-16 Nov. 2007.

Zhang H., Siu K.Y., Li A.S.M., Chan K.Y.Q., Ngan H.Y.S. and Cheung A.N.Y., Oct-4 gene is epigenetically regulated by methylation in normal placenta and gestational trophoblastic disease, 14th Hong Kong International Cancer Congress, Hong Kong, 14-16 November. 2007.

Zhang H., Siu K.Y., Wong E.S.Y., Wong K.Y., Li A.S.M., Chan Y.K., Ngan H.Y.S. and Cheung A.N.Y., Oct4 is Epigenetically Regulated by Methylation in Normal Placenta and Gestational Trophoblastic Disease, Placenta. 2008, 29(6): 549-554.

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