Research Into Digestive Diseases Digestive Into Research

tumorigenicity. The mutant protein also exhibited altered altered exhibited also protein The mutant tumorigenicity. Whole-exome sequencing of 10 colon cancer patients identified a Single-cell whole-exome sequencing has been carried out to catalogue novel recurrent mutation. This mutation was detected in 23 out of 118 patients (20.18%) in the validation cohort. Ectopic expression of this mutant in colon cancer cells increased cell proliferation and colony- forming ability, caused accumulation of cells in S-phase, and enhanced in vivo subcellular localisation. somatic mutations in 63 cancer cells isolated from a colon cancer somatic mutations in 63 cancer cells isolated from a colon cancer specimen. The mutation spectrum was heterogeneous at the single- cell level. The accumulation of mutations was closely related to tumorigenesis based on population genetic analysis. Among them, we identified a high-frequency mutated gene at the single-cell level, which showed low prevalence in an additional cohort study of colon cancer. potential its revealed gene mutant the of characterisation Functional oncogenic effect in colon cancer.

2. RESEARCH PROGRESS SUMMARY: CANCER COLORECTAL A. Molecular Pathogenesis 1.

Research into Digestive Diseases Digestive into Research

PI: Team: Joseph Sung, Henry Chan, Vincent Wong, Justin Wu, Siew Ng (Department of Medicine & Therapeutics, Institute of Digestive Diseases), Cheng, William Alfred Wu (Institute of Digestive Diseases), Enders Ng, James Lau, Simon Ng, Philip Chiu (Department of Surgery, Institute of Digestive Diseases) Francis Chan, Jun Yu Francis Chan, Jun Yu (Department of Medicine & Therapeutics, Institute of Digestive Diseases)

Technical Report on Research Progammes 44 45 01 Research into Digestive Diseases

YY1 analysis revealed that patients with patients that revealed analysis Colitis is associated with increased risk for colorectal Colitis is associated with increased risk Multivariate Multivariate demonstrated marginal Cap-assisted colonoscopy colorectal cancer were found Siblings of patients with A cancer. Cathelicidin, a pleiotropic peptide, was was peptide, cancer. Cathelicidin, a pleiotropic in mice. The found to alleviate experimental colitis cytokines, altered tissue levels of pro-inflammatory mucus and apoptosis, activity, myeloperoxidase by reversed be can colitis with mice in secretion cathelicidin- intrarectal administration of cathelicidin or encoding plasmid. benefit over standard colonoscopy for polyp detection detection polyp for colonoscopy standard over benefit intubation time. and shortened the caecal of advanced neoplasms to have a higher prevalence cancers or adenomas of at least(defined as colorectal high-grade dysplasia, with villous 10 mm in diameter, healthy of siblings than characteristics) tubulovillous or high-risk this in indicated is Screening individuals. population. and 37 induces caspase-independent apoptosis p53- autophagic cell death through the common Our study Bcl-2/Bax cascade in colon cancer cells. previously unknown reciprocal also uncovered pathways. regulation between these two cell death protein high expression had a significant decrease in had a significant decrease in protein high expression overall survival. 2. 2. 3. 4. 1. LL- peptide suppressing tumour the of fragment C. Molecular Therapeutics

Our We identified that miR-7, a microRNA downregulated We identified that miR-7, a microRNA downregulated Hydrogen sulphide, a gaseous bacterial metabolite Hydrogen sulphide, A samples as potential screening biomarkers for for samples as potential screening biomarkers colorectal colorectal cancer and polyps. Patients with miR-21 level cancer had a significantly higher stool controls. and miR-92a level compared with normal significantly Stool miR-92a, but not miR-21, was a At controls. in than polyps with patients in higher miR-92a cut-off value of 435 copies/ng of stool RNA, for colorectal had a sensitivity of 71.6% and 56.1% specificity of cancer and polyp, respectively, and a 73.3%. in colorectal cancer, suppresses colon tumorigenesis in colorectal cancer, suppresses colon tumorigenesis ). by targeting the oncogenic Yin Yang 1 (YY1 that reaches high levels in the large intestine, was in the large intestine, was that reaches high levels and induces protective reported to lower proliferation epithelial cells via activation of autophagy in colon kinase protein monophosphate-activated adenosine signalling. by macrophages, polymorphonuclear leukocytes, leukocytes, polymorphonuclear macrophages, by reported to contribute to colon and colonocytes was by activating a novel pathway of cancer suppression Factor Apoptosis-Inducing by mediated apoptosis colon cancer in (EndoG) G (AIF) and Endonuclease cells.

1. stool in miR-21 and miR-92a identified study 5. 4. 3. secreted cathelicidin as known factor bactericidal B. Biomarkers and Cancer Screening

agonist agonist γ γ as a valid as a valid ) activation ) activation γ γ PPAR PPAR PPAR through regulating key through regulating key exerts an inhibitory effect onexerts an inhibitory effect γ in vivo PPAR in vitro and in vivo, thus representing the invasive and metastatic potential of hepatocellular the invasive and metastatic carcinoma (HCC) and treatment of HCC a target for the prevention metastasis. F W We have previously demonstrated that peroxisome previously demonstrated We have miR-139, down-regulated in We delineated that inhibits hepatocarcinogenesis. We further We further hepatocarcinogenesis. inhibits demonstrated that in research obese patients, we are actively engaging a (FZHY), recipe Huayu Fuzheng therapy. NASH of was reported compound of Chinese herbal medicine, with patients in fibrosis and function liver improve to the hepatitis B virus infection. We demonstrated nutritional protective role of FZHY in ameliorating fibrosing NASH and genes related to oxidative stress, inflammation fibrogenesis. proliferator-activated receptor ( receptor proliferator-activated suppresses HCC migration by metastatic HCC cells, c-fos. targeting the oncogenic in preventing HCC development and spread using in preventing HCC development and spread using pre-clinical models, thus pointing therapeutic target against HCC.

1. in prevalent more is which HCC of control the or 2. of effects crucial the confirmed e A. Molecular Pathogenesis A. Molecular 1. 2. LIVER CANCER B. Biomarkers and Cancer Screening B. Biomarkers and (NASH), anThe diagnosis of non-alcoholic steatohepatitis by the need for emerging risk factor for HCC, is limited combined serum liver biopsy. We tested the accuracy of of NASH. A biomarkers for the non-invasive diagnosis and fragment cytokeratin-18 using approach two-step to accurately fibroblast growth factor 21 was reported diagnose NASH. C. Molecular Therapeutics

B (Hp) (Hp) were were RNF180 RNF180 ZNF545 ZNF545 and and ZNF545 ), Zinc-finger protein ), Zinc-finger protein RNF180 that hypermethylated tumour- that HOXD10, RNF180 HOXD10, BCL6B, PAX5 BCL6B, PAX5 RNF180 DNA was detected in the plasma

331 (ZNF331) and ZNF545 was reported to activate to activate 331 (ZNF331) and ZNF545 was reported cancer. growth and anti-apoptotic pathways in gastric critical for Besides, we demonstrated that genes e.g. normal cell differentiation and development cell CLL/lymphoma 6 member B (BCL6B), HOXD10, HOXD10, cell CLL/lymphoma 6 member B (BCL6B), gastric in involved were , (PAX5) 5 gene box paired cancer progression when aberrantly methylated. W We demonstrated Novel epigenetic aberrations were found to occur Novel epigenetic aberrations were found Using genome-wide methylation screening, we Using genome-wide We discovered that major aetiological microorganisms microorganisms that major aetiological We discovered associated with shortened survival in different stages associated with shortened survival in different stages of gastric cancer patients. suppressor genes as independent biomarkers for suppressor genes as independent biomarkers for prognosis of gastric cancer patients. For example, methylation of early in gastric carcinogenesis. For example, promoter early in gastric carcinogenesis. For example, of methylation identified potential tumour-suppressor genes that genes that identified potential tumour-suppressor play important roles in gastric carcinogenesis. genes, Silencing of these novel tumour-suppressor such as previously known as hypothetical proteins ring finger protein 180 ( and Epstein-Barr virus (EBV) cause epigenetic cause epigenetic (EBV) virus and Epstein-Barr carcinogenesis. In particular, dysregulation to promote transcriptional forkhead box D3 (FOXD3)-mediated is deregulated by Hp control of tumour suppressors which in turn infection-induced hypermethylation, between cell death and survival. perturbs the balance factor 1 (ZEB1) was reportedZinc finger E-box binding the latent-lytic switch of EBV- as a key mediator of FOXD3 and ZEB1 may be associated gastric cancer. infection-induced gastric cancer potential targets for therapy. Helicobacter pylori cancer namely Helicobacter of gastric methylation for screening gastric cancer patients. patients. cancer gastric screening for methylation Methylated of ~60% of gastric cancer patients, but not in healthy controls. was detected in 30-60% of intestinal metaplasia, a metaplasia, a was detected in 30-60% of intestinal findings precancerous lesion of gastric cancer. These cancer diagnosis new strategies for promising provide and prevention.

2. of utility clinical potential the explored e 1. 3. 2. A. Molecular Pathogenesis A. Molecular 1. GASTRIC CANCER GASTRIC C. Molecular Therapeutics In addition to tumour-suppressor gene silencing, we found that up-regulation of novel oncogenes activates in gastric cancer. Stathmin1 signalling pathways key prognosis and oncoprotein candidate a is (STMN1) marker in several kinds of cancers. We confirmed that miR-223 of target downstream putative a is STMN1 in gastric cancer. Our findings revealed that STMN1 a as serve might and oncogene gastric potential a is therapeutic target for gastric cancer. B. Biomarkers and Cancer Screening

Technical Report on Research Progammes 46 47 01 Research into Digestive Diseases Founding Fellow of Wu Yee Sun College, The Chinese University of Hong Kong Visiting Professor, Shanghai Medical College, Fudan University (July 2012 - July 2014) Distinguished Research Paper Award for Young Investigators, Hong Kong College of Distinguished Research Paper Award for Physicians (October 2012) Digestive Week (December 2012) Emerging Leader Lectureship, Asian Pacific American the to Fellowship Traveling (ESCP) Coloproctology of Society European 2012) Society of Colon and Rectal Surgeons (ASCRS) Annual Scientific Meeting (June College Best Scientific Paper Award of the Conjoint Scientific Congress of The Royal of Surgeons of Edinburgh and The College of Surgeons of Hong Kong (September 2012) First Prize of the World up of Endoscopy, American Society of Gastrointestinal Endoscopy, United States of America Professor Richard Yu Medal, Advances in Medicine 2012 Professor Richard Yu Medal, Advances in (May Kong Hong of University Chinese The - 12, 2011 Award Excellence Research 2012) United European Gastroenterology Week Travel Grant Award for Basic Scientists, (October 2012) Gastroenterology Week (October 2012) Oral Free Paper Prize, United European Foundation, Disease Digestive Prize, Abstract Gastroenterology Frontline Inaugural United Kingdom (June 2012) National Science & Technology Progress Award 2012 National Science & Technology of the Higher Education Progress Award, Class 1, Scientific and Technological Ministry of Education, Research Output Awards 2012, The Outstanding Scientific China Institute of Advanced Academy of Sciences, Guangzhou Visiting Professor, Chinese 2012 - November 2014) Technology (December 2, of the Higher Education Outstanding Technological Advancement Award, Class The Ministry of Education, China Scientific Research Output Awards 2011,

• • • • • • • • • • • • • • • • Details Jun Yu Siew Ng Simon Ng Philip Chiu Philip Chiu Alfred Cheng Francis Chan Francis Chan, Joseph Sung, Vincent Wong Jun Yu, Joseph Jun Yu, Vincent Member’s Name Member’s Sung, Henry Chan, Joseph Sung, et. al Wong, Henry Chan, Vincent Wong, et. al. AWARDS AND FELLOWSHIPS AWARDS RECOGNITIONS: 998,984 891,776 342,000 447,132 247,274 368,490 685,020 589,493 994,040 241,500 1,380,000 Amount (HK$) Name Jun Yu Siew Ng Siew Ng Simon Ng Simon Ng Philip Chiu Member’s Member’s William Wu James Lau Alfred Cheng Francis Chan Vincent Wong

virus-driven novel promoter hypermethylated genes (SSTR1, virus-driven novel promoter hypermethylated genes (SSTR1, REC8, IL15RA) in gastric cancer. (HBx)-mediated liver inflammation and carcinogenesis. on acupoints (Acu-TENS) for pain relief during colonoscopy: a on acupoints (Acu-TENS) for pain relief during colonoscopy: a prospective, randomized, placebo-controlled study. B: a prospective study with paired transient elastography examination. positive subjects with familial adenomatous polyposis. TNF therapy in inflammatory bowel disease. peptic ulcers after their initial endoscopic hemostasis – a hemostasis – a peptic ulcers after their initial endoscopic randomized controlled trail. for the relief of upper gastrointestinal obstruction in patients in patients the relief of upper gastrointestinal obstruction for upper gastrointestinal suffering from advanced malignant diseases. virus-associated hepatocarcinogenesis. misoprostol for healing of small bowel ulcers in aspirin users with of small bowel ulcers in aspirin users misoprostol for healing small bowel bleeding. Functional significance and clinical application of Epstein-Barr Elucidating the role of autophagy in hepatitis B virus X protein Liver fibrosis progression in patients with chronic hepatitis A phase III placebo-controlled trial of Cclecoxib in genotype The application of transcutaneous electric nerve stimulation R study (ACCESS). Asia-pacific Crohn's and colitis epidemiology Early selective angiographic embolization to severely bleeding Early selective angiographic embolization Grant for the use of self-expandable-metallic-stents (SEMS) Grant for the use of self-expandable-metallic-stents A novel androgen receptor oncogenic circuitry in hepatitis B oncogenic circuitry in hepatitis A novel androgen receptor A double-blind, randomized, placebo controlled trial of placebo controlled trial A double-blind, randomized,

Research Fund for the Control of Infectious Diseases (RFCID) (RFCID) Diseases Infectious of Control the for Fund Research (2012 - 2014) Health and Medical Research Fund (HMRF) (2012 - 2014) Health and Medical Research Title: Title: Title: Research Grants Council (RGC) General Research Fund (GRF) (GRF) (RGC) General Research Fund Research Grants Council (2012 - 2015) Title: Title: Title: anti- during tuberculosis for monitoring in quantiferon of ole Title: Title: Title: Title: Details Title: Pfizer Corporation Hong Kong Limited, Hong Kong (2010 - 2013) Pfizer Corporation Hong Kong Limited, Ferring, Hong Kong (2011 - 2013) Health and Medical Research Fund (HMRF) (2011 - 2012) (RFCID) Diseases Infectious of Control the for Fund Research (2012 - 2014) SK Yee Medical Foundation (2012 - 2014) (RFCID) Diseases Infectious of Control the for Fund Research (2012 - 2014) (RFCID) Diseases Infectious of Control the for Fund Research (2012 - 2013) Research Grants Council (RGC) General Research Fund (GRF) Research Fund (GRF) Research Grants Council (RGC) General (2010 - 2012)

Grants GRANTS AND CONSULTANCY GRANTS

Technical Report on Research Progammes 48 49 01 Research into Digestive Diseases 851,627 RMB 80,000 RMB 5,000,000 RMB 12,000,000 Jun Yu Jun Yu Jun Yu (Co-I) Jun Yu (Co-I)

Advisory Board Member of Abbott, Bristol Myers Squibb, FuRui, Gilead, Merck, Myers Squibb, FuRui, Gilead, Merck, Advisory Board Member of Abbott, Bristol Novartis, Roche, Vertex Advisor, Gilead Pharmaceuticals Hong Kong Honorary Consultant, Regeneration Society,

• • • Details on expression, function, and clinical application of DACT1 of application clinical and function, expression, on 重大疾病的基因組學技術腸道微生態與感染及代謝的研究 cancer in stool. in gastric cancer. A novel approach for the non-invasive diagnosis of colorectal diagnosis for the non-invasive A novel approach Study

Henry Chan Vincent Wong Vincent Wong Member’s Name Member’s Shenzhen Basic Research Program, China (2012 - 2013) Shenzhen Basic Research National 863 Program, China (2011 - 2015) National 863 Program, China (2012 - 2017) National 863 Program, Title: Title: Title: Title: Innovation and Technology Fund, Hong Kong (2012 - 2013) Hong Kong (2012 and Technology Fund, Innovation Consultancy

, Gut PLoS PLoS Cancer Cancer , 86(7), 86(7), , Journal of of Journal , 27(11), 1665- British Journal of British Journal of . in mice Journal of Virology and Journal of Gastroenterology and and Gastroenterology of Journal in vitro in hepatocellular carcinoma. carcinoma. hepatocellular in γ 61(5), 739-745. Leung, W. W., Lee, C. W., Wong, Y. N., Chan, F. K., Leung, W. W., Lee, C. W., Wong, Y. N., of miR-92a Yu, J., & Sung, J. J. (2012). Detection and miR-21 in stool samples as potential screening biomarkers for colorectal cancer and polyps. J. J. Sung, Yu, J., & K., W., Chan, F. C. Lee, therapy. cancer in paradox autophagic The (2012). Oncogene, 31(8), 939-953. Xu, L., Li, X., Chu, E. S., Zhao, G., Go, M. Y., Tao, Q., Jin, H., Zeng, Z., Sung, J. J., & Yu, J. (2012). Epigenetic inactivation of BCL6B, a novel functional tumour suppressor for gastric cancer, is associated with poor survival. Gut, 61(7), 977-985. Cancer, 106(9), 1486-1494. G. C., & Yu, J. (2012). Wu, C. W., Farrell, Functional role of peroxisome-proliferator-activated receptor Gastroenterology and Hepatology 1669. Ng, S. C., Wu, C. W., Ng, S. S., Dong, Y. J., Wu, C. W., & Yu, J. (2012). Heme oxygenase-1 organ in regime anti-inflammatory the induction: transplant. Hepatology, 27(4), 621-622. X. J., C. H., Wang, Wu, W. K., Coffelt, S. B., Cho, S., A. K., Cheng, F. Chan, L., Yu, C., Wang, X. J., Y. Wu, Yu, J., Sung, J. J., Wu, W. K., & Cho, C. H. (2012). Hydrogen sulfide lowers proliferation and induces protective autophagy in colon epithelial cells. One, 7(5), e37572. Lu, D., Wu, Y., Wang, Y., Ren, F., Wang, D., Su, F., F., Wang, D., Su, F., Y., Wang, Y., Ren, Lu, D., Wu, X., He, D., Zhai, Y., Yang, X., Jin, G., Hao, Zhang, Y., & B., Jia, J., Yu, J., J. Sung, J., Hu, M., D. Irwin, qccelerates tumorigenesis Chang, Z. (2012). CREPT cell-cycle-related of transcription the regulating by genes. Cancer Cell, 21(1), 92-104. J., Yu, J., Chen, Y., Lin, M. C., Lu, J., Yi, L., Zhao, 71 Enterovirus (2012). L. M. He, & F., H. Kung, level the reducing by signaling interferon disrupts 1. receptor interferon A. S., To, K. F., Tong, J. H., Ren, S. X., Cheng, C. C., Zhang, L., Chan, R. L.,Li, M. S., Shen, J., Wong, V. E., Marquez, L. C., S., Chiu, X. J., Ng, S. Wang, J., J. Sung, J., Yu, K., F. Chan, L., R. Gallo, defense Wu, W. K., & Cho, C. H. (2012). Host immune caspase-independent activates LL-37 peptide apoptosis and suppresses colon cancer. Wu, C. W., Shen, B., Chu, E. S., Zhao, G., Man, K., Teoh, N., Cheng, J. T., Li, G., Nie, Y., Lo, C. M., (2012). J. Yu, & J., J. Sung, C., G. Farrell, carcinoma PPARgamma inhibits hepatocellular metastases 3767-3776. Research, 72(24), 6512-6523.

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, , (2012). (2012). PLoS One et al. , 30(4), 271- , , a novel gene in gastric gastric , a novel gene in Lipids in Health and Disease RNF180 Cell BiochemistryCell and Function

Liu, Y., Song, F., Wu, W. K., He, M., Zhao, L., Sun, X., Liu, Y., Song, F., Wu, W. K., He, M., Zhao, L., Sun, Li, Z., Shen, J., Wu, W. K., Wang, X., Liang, J., Qiu, G., Li, Z., Shen, J., Wu, W. K., Wang, X., Liang, J., Qiu, G., & Liu, J. (2012). Vitamin A deficiency induces congenital spinal deformities in rats. Li, Z., Shen, J., Wu, W. K., Yu, X., Liang, J., Qiu, G., & Liu, J. (2012). Leptin induces cyclin D1 expression and proliferation of human nucleus pulposus cells pathways. MEK/ERK and PI3K/Akt JAK/STAT, via PLoS One, 7(12), e53176. Guidelines for the use and interpretation of assays for for assays of interpretation and use the for Guidelines monitoring autophagy. Autophagy, 8(4), 445-544. J., Zhao, L., Tian, X., Ma, F., K. Cheung, X., Li, J., Tao, Q., Go, M. Y., Shen, B., Cheng, A. S., Ying, Sung, J. J., Kung, H. F., & Yu, J. (2012). Epigenetic inactivation of paired box gene 5, a novel tumor suppressor gene, through direct upregulation of p53 is associated with prognosis in gastric cancer patients. Oncogene, 31(29), 3419-3430. 7(10), e46565. Li, H., Jiang, Y., Yang, Y., & Peng, K. (2012). Triptolide inhibits colon cancer cell proliferation and induces cleavage and translocation of 14-3-3 epsilon. 278. Klionsky, D. J., Sung, J. J., Wu, W. K., D. J., Sung, Klionsky, Kang, W., Tong, J. H., Chan, A. W., Lung, R. W., R. W., Kang, W., Tong, J. H., Chan, A. W., Lung, Cheng, A. S.,Chau, S. L., Wong, Q. W., Wong, N., Yu, J., role & To, K. F. (2012). Stathmin1 plays oncogenic cancer. gastric in microRNA-223 of target a is and PLoS One, 7(3), e33919. Jia, Y. H., Wang, R .Q., Mi, H. M., Kong, L. B., L. B., Jia, Y. H., Wang, R .Q., Mi, H. M., Kong, Chong, W. W., Cheng, A. S., To, K. F., Fan, D., To, K. F., Fan, D., W., Cheng, A. S., Chong, W. the of J. (2012). Characterization & Yu, J. J., Sung, significance, and clinical gene structure, functional application of J. (2012). Co-inhibition of Shao, J. F., & Qiao, microRNA-21 exerts synergistic microRNA-10b and and invasion of human inhibition on the proliferation of Oncology, 41(3),glioma cells. International Journal 1005-1012. C. G., Wu, W. K., Wang, X. J., Feng, S. Y., Dong, expression Qiao, J., & Shao, J. F. (2012). Lentiviral inhibits miR-27a targeting anti-microRNAs of cells. proliferation and invasiveness of U87 glioma Molecular Medicine Reports, 6(2), 275-281. Y. G., Ren, W. G., Li, W. C., Zhao, S. X., Zhang, Fuzheng Wu, W. J., Nan, Y. M., & Yu, J. (2012). fibrosing Huayu recipe prevents nutritional steatohepatitis in mice. 11(1), 45. cancer. Cancer, 118(4), 947-959. J., X. K., Feng, S. Y., Wang, Dong, C. G., Wu, W. Cheung, K. F., Lam, C. N., Wu, K., Ng, E. K., K., E. Ng, K., Wu, C. N., K. F., Lam, Cheung,

10. 8. 9. 7. 6. 5. 4. 3. 2. PUBLICATIONS: 1.

Technical Report on Research Progammes 50 51 01 Research into Digestive Diseases

as an imaging biomarker for biomarker for as an imaging ρ Zhao, J., Jin, H., Cheung, K. F., Tong, J. H., Cheung, K. F., Tong, J. H., Zhao, J., Jin, H., Zhao, F., Wang, Y. X., Yuan, J., Deng, M., Wong, H. L., H. Wong, M., Deng, J., Yuan, X., Y. Wang, F., Zhao, & G. J., Ahuja, A. T., Go, M. Y., Teng, Chu, E. S., MR T1 Yu, J. (2012). an regression: and progression injury liver monitoring rats with carbon tetrachloride experimental study in , 22(8), 1709-1716. intoxication. European Radiology Zhang, S., Go, M. Y., Tian, L., Kang, W., Leung, P. P.,Zhang, S., Go, M. Y., F., Sung, J. J., & Yu, J. (2012).Zeng, Z., Li, X., To, K. central a plays 1 factor binding E-box finger Zinc virus (EBV) latent- role in regulating Epstein-Barr as a therapeutic target in EBV- lytic switch and acts Cancer, 118(4), 924-936. associated gastric cancer.