1. Personal Information 1 10

2. 2 10 Teh-Ying Chou M.D., National Yang-Ming Medical College ; Ph.D., Johns Hopkins University School of Medicine / 66/9/1~73/6/31 75/9/1~79/8/31 / 84/4/1~84/6/30 Johns Hopkins 84/7/1~85/6/30 Johns Hopkins 85/8/28~95/1/31 85/12/16~88/6/30 88/7/1~90/6/30 Washington University 90/7/1~91/6/30 Armed Forces Institute of Pathology (AFIP) (Calendar Binford Fellow) 91/10/1~93/10/31 93/11/1~95/7/31 :02-28757080 (office) 02-28757449 ext.295 (Lab) Fax:02-28757056 e-mail tychou@vghtpe.gov.tw 3. Research Topics Invasion and Metastasis of Lung Cancer Thoracic Pathology Having been trained in a Biochemistry, Cellular and Molecular Biology Ph.D. Program and the subspecialty of Pulmonary Pathology, and practicing as a pulmonary pathologist, I chose to establish my laboratory with a research theme of pathobiology of lung diseases. Research activities in my laboratory are summarized below: (1) Collecting high-quality lung cancer specimens for the forthcoming studies: Supported by the National Health Research Institutes (Grants NHRI92A1- NSCCO-23-5 and NHRI93A1-NSCCO-23-5) and in collaboration with chest surgeons, we have established a pathology-based tissue bank for both tumorous and non-tumorous lung tissues. Since April 2003, cases of high-quality lung cancer specimens including blood samples and freshly frozen lung tissues have been successfully collected. Together with the patients clinical information, the detailed pathological diagnosis and description of each specimen were recorded and stored. Our standard tissue procurement protocol for fresh lung cancer specimens may serve as a model scheme for other lung cancer tissue banks in Taiwan. The specimens from our tissue collection have been proven to be of high quality, as we were able to successfully perform nucleic acid and protein extractions as well as in situ molecular pathology studies using these samples. These tissues provide superb materials for our proposed projects. (2) EGFR-TK mutations in pulmonary adenocarcinomas: In a collaborative work involving chest physicians, basic researchers and pathologists, we have investigated the association between mutations in the EGFR-TK domain and tumor response and survival in 54 Gefitinib-treated NSCLC patients. Genomic DNA was prepared from formalin-fixed, paraffin-embedded sections of tumor specimens. Eight pairs of oligonucleotide primers were used to amplify exons 18, 19, 20 and 21 of the EGFR gene by nested PCR, and the PCR products were subsequently sequenced. Our results showed that EGFR mutations occurred in 61.1% of cases and the presence of 3 10 4. EGFR mutations was the only independent predictor for disease control (P= 0.003) and tumor response (P= 0.017) by Gefitinib treatment. In comparison with patients whose tumor was negative for EGFR mutations, patients with EGFR mutations had better progression-free survival and overall survival. Our study demonstrated that mutations in EGFR-TK could be used as a predictive and prognostic factor in Gefitinib-treated NSCLC patients. This study has yielded a publication entitled Mutation in the Tyrosine Kinase Domain of Epidermal Growth Factor Receptor Is a Predictive and Prognostic Factor for Gefitinib Treatment in Patients with Non-Small Cell Lung Cancer (Clin Cancer Res 11(10):3750-3757, 2005). (3) Genetic profiling of in situ, invasive and metastatic pulmonary adenocarcinomas: Supported by the Taipei Veterans General Hospital (Grant VGH- 94-286), we have performed a subtractive hybridization experiment using primary tumor and metastatic tumor from a female patient of pulmonary adenocarcinoma, obtained and sequenced approximately 150 metastasis-associated candidate clones that were expressed at higher levels in the metastatic tumor. We are currently in the process of verifying the differential expression of these metastasis-associated genes (which include Clusterin) in pulmonary adenocarcinomas and exploring their molecular mechanisms in tumor metastasis. In addition, supported by the National Science Council (Grants NSC94-2314-B-010-059 and NSC95-2314-B-010-027), we have initiated a pilot study using laser capture microdissection of in situ tumor cells and invasive tumor cells from a male patient of pulmonary adenocarcinoma. We obtained from cDNA microarray analysis approximately 45 genes with 3-fold up- regulation in invasive tumors as compared to in situ tumors, which included matrix metalloproteinases, a few known metastasis-associated genes, and some candidates for novel invasion-associated genes. Among the novel ones, the differential expression of prion protein (PrPc), which is a membrane glycosylphosphatidylinositol-anchored glycoprotein well known for its implication in Bovine Spongiform Encephalopathy (BSE) and Creutzfeld-Jakob Disease (CJD), has been confirmed using RT-PCR and immunohistochemical methods. The clinical correlation of differential expression and the role of prion protein in the mechanism of invasion process are currently being explored in cell line systems (manuscript in preparation). (4) Clusterin silencing induces a mesenchymal-to-epithelial transition in human lung adenocarcinoma cells: Clusterin (CLU) is implicated in diverse cellular processes, yet its genuine molecular function remains enigmatic. We have discovered CLU as a metastasis-associated gene by subtractive hybridization and cDNA microarray analysis; CLU was differentially expressed at higher levels in metastatic 4 10 5. than in the primary lung adenocarcinoma tissues. In human lung adenocarcinoma cell lines with different invasive capabilities, CLU levels positively correlated with the degree of invasiveness. Moreover, high CLU-expressing cells displayed a mesenchymal morphology while cells with less CLU were epithelioid in shape, suggesting the possibility that CLU drives epithelial-mesenchymal transitions (EMT) in human lung adenocarcinoma cells. Consistently, silencing of CLU by siRNA in the highly invasive, high CLU-expressing lung adenocarcinoma cell line resulted in a morphological change from mesenchymal to epithelioid shapes, increased E-cadherin expression, and decreased fibronectin expression. CLU-knockdown cells, compared to the vector-transfected cells, were inhibited in migration and invasion in wound healing assay and matrigel invasion assays, respectively. In an experimental metastasis assay, mice injected with CLU-knockdown lung adenocarcinoma cells did not develop lung nodules as those injected with vector-transfected cells. Re- expression of CLU in CLU-knockdown cells restored the mesenchymal and invasive phenotypes, suggesting that lack of CLU function caused the MET and decreased migration and invasiveness in CLU-knockdown cells. Furthermore, we examined a panel of transcriptional repressors of E-cadherin and found that Slug expression was decreased in CLU-knockdown cells. We also discovered that levels of phosphorylated ERK correlated with those of CLU and Slug. Taken together, our data suggest that CLU may modulate EMT and invasive phenotypes in human lung adenocarcinoma cells through ERK and Slug signaling. This study has yielded a publication entitled Clusterin silencing in human lung adenocarcinoma cells induces a mesenchymal-to- epithelial transition through modulating the ERK/Slug pathway (Cellular Signalling 21(5):704-711, 2009). (5) Decoy Receptor 3 Levels in Peripheral Blood Predict the Outcome of Patients with Acute Respiratory Distress Syndrome: Acute respiratory distress syndrome (ARDS), a serious inflammatory reaction to acute lung injury, is associated with high mortality rates. Biomarkers that reliably predict outcomes in ARDS are not currently available. Decoy receptor 3 (DcR3) is a soluble protein with immunomodulatory effects. The role of DcR3 in ARDS is unknown. Comparing DcR3 with the APACHE II scores and three other plasma markers, we explored the association of DcR3 and the clinical outcome in ARDS. Eighty-eight ARDS patients were studied. Baseline APACHE II scores and clinical data were recorded. Plasma levels of DcR3, soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), tumor necrosis factor (TNF), and interleukin-6 (IL-6) were measured on Day 1 and later time points, and correlated with the survival status on Day 28 after the onset of ARDS. Among the four biomarkers evaluated, only DcR3 discriminated the survivors 5 10 6. and non-survivors at all time points in the first week of ARDS development. DcR3 was independently associated with and best predicted the 28-day mortality of ARDS patients. Plasma DcR3 levels most correlated to multiple-organ dysfunction and ventilator dependence. Regardless of either low or high APACHE II scores, Day 1 DcR3 levels were significantly higher in the non-survivor group of patients. Kaplan- Meier survival analysis showed higher mortality in ARDS patients with higher DcR3 levels. A high plasma DcR3 level independently predicts the 28-day mortality of patients with ARDS. This study has yielded a manuscript entitled Decoy Receptor 3 Levels in Peripheral Blood Predict the Outcome of Patients with Acute Respiratory Distress Syndrome (American Journal of Respiratory and Critical Care Medicine 180(8):751-760, 2009 ). Publications 1. Chen CY, Yang KY, Chen MY, Chen HY, Lin MT, Lee YC, Perng RP, Hsieh SL, Yang PC, Chou TY. * Decoy Receptor 3 Levels in Peripheral Blood Predict Outcomes of Acute Respiratory Distress Syndrome. American Journal of Respiratory and Critical Care Medicine (2009) 180(8):751-760(SCI)(Impact factor: 9.792;Rank:1/40, 2.50%)(* Corresponding Author) 2. Yeh YC, Chou TY. * Pulmonary adenocarcinoma with microcystic histology and intratumoral heterogeneity of EGFR gene polymorphism. Histopathology (2009) [Epub ahead of print]. (SCI)(Impact factor: 4.131;Rank:10/69, 14.49%)(* Corresponding Author) 3. Chou TY,* Chen WC, Lee AC, Hung SM, Shih NY, Chen MY.* Clusterin silencing in human lung adenocarcinoma cells induces a mesenchymal-to- epithelial transition through modulating the ERK/Slug pathway. Cellular Signalling (2009) 21(5):704-711(SCI)(Impact factor: 4.305;Rank: 52/157, 33.12%)(* Corresponding Author) 4. Yang MH, Chen CL, Chau GY, Chiou SH, Su CW, Chou TY, Peng WL, Wu JC. Comprehensive analysis of the independent effect of twist and snail in promoting metastasis of hepatocellular carcinoma. Hepatology (2009) 50(5):1464- 1474(SCI) (Impact factor: 11.355;Rank:2/55, 3.64%) 5. Yen CH, Hung JH, Ueng YF, Liu SP, Chen SY, Liu HH, Chou TY, Tsai TF, Darbha R, Hsieh LL, Chen YM. Glycine N-methyltransferase affects the metabolism of aflatoxin B(1) and blocks its carcinogenic effect. Toxicology and applied Pharmacology (2009) 235(3):296-304 (SCI)(Impact factor: 3.364;Rank: 11/75, 14.67%) 6. Lin CS, Wang LS, Chou TY, Chang SC, Hsu WH, YH Wei. The role of 6 10 7. mitochondrial DNA alterations in esophageal squamous cell carcinomas. Journal of Thoracic and Cardiovascular Surgery (2009) [Epub ahead of print]. (SCI) (Impact factor: 3.037;Rank: 20/148, 13.51%) 7. Kuo KT, Wang HW, Chou TY, Hsu WH, Hsu HS, Lin CH, Wang LS. Prognostic Role of PGE2 Receptor EP2 in Esophageal Squamous Cell Carcinoma. Annals of surgical oncology (2009) 16(2):352-360 (SCI)(Impact factor: 3.898;Rank: 9/148, 6.08%) 8. Hu YH, Hsiao LT, Yang CF, Chiou TJ, Liu JH, Gau JP, Yen CC, Chou TY, Hsu WH, Chen PM, Tzeng CH. Prognostic factors of Chinese patients with primary pulmonary non-Hodgkin's lymphoma: the single-institute experience in Taiwan. Annals of hematology (2009) 88(9):839-846 (SCI)(Impact factor: 2.454;Rank: 30/62, 48.39%) 9. Chen CH, Lai JM, Chou TY, Chen CY, Su LJ, Lee YC, Cheng TS, Hong YR, Chou CK, Whang-Peng J, Wu YC, Huang CY. VEGFA upregulates FLJ10540 and modulates migration and invasion of lung cancer via PI3K/AKT pathway. PLoS One (2009) 4(4):e5052 (SCI) 10. Lee HT, Su WJ, Chou TY, Chen WS, Su KY, Huang DF. Mycobacterium Avium Complex-Induced Pleurisy in a Patient With Amyopathic Dermatomyositis and Interstitial Lung Disease After Prolonged Immunosuppressive Therapy. JCR- Journal of Clinical Rheumatology (2009) 15(4):193-194. (Impact factor: 1.416;Rank: 18/22, 81.82%) 11. Lin CS, Chang SC, Wei YH, Chou TY, Wu YC , Lin HC, Wang, LS, Hsu WH. Prognostic Variables in Thoracic Esophageal Squamous Cell Carcinoma. Annals of thoracic surgery (2009) 87:1056-1065. (Impact factor: 2.689;Rank: 23/148, 15.54%) 12. Wu MF, Yang CY, Lin TL, Wang JT, Yang FL, Wu SH, Hu BS, Chou TY, Tsai MD, Lin CH, Hsieh SL. Humoral immunity against capsule polysaccharide protects host from magA+ Klebsiella pneumoniae-induced lethal disease through evading TLR4 signaling. Infection and immunity (2009) 77(2):615-621 (SCI) (Impact factor: 3.987;Rank: 10/51, 19.61%) 13. Hung JJ, Chou TY, Sun CH, Liu JS, Hsu WH. Primary Synovial Sarcoma of the Posterior Chest Wall. Annals of thoracic surgery (2008) 85:2120-2122 (SCI) (Impact factor: 2.689;Rank: 23/148, 15.54%) 14. Chen YM, Liu JM, Chou TY, Perng RP, Tsai CM, Whang-Peng J. Phase II randomized study of daily gefitinib treatment alone or with vinorelbine every 2 weeks in patients with adenocarcinoma of the lung who failed at least 2 regimens of chemotherapy. Cancer (2007) 109(9):1821-1828 (SCI)(Impact factor: 5.238;Rank: 19/141, 13.48%) 7 10 8. 15. Wu MH, Chern MS, Liu JC, Sheu MH, ChouTY, Chang CY. Massive air leak after CT-guided lung biopsy. European Journal of Radiology (2006) 60(1):15- 17 (SCI) (Impact factor: 2.339;Rank: 36/90, 40.00%) 16. Chang GC, Liu KJ, Hsieh CL, Hu TS, Charoenfuprasert S, Liu HK, Luh KT, Hsu LH, Wu CW, Ting CC, Chen CY, Chen KC, Yang TY, Chou TY, Wang WH, Whang-Peng J, Shih NY. Identification of alpha-enolase as an autoantigen in lung cancer: its overexpression is associated with clinical outcomes. Clinical Cancer Research (2006) 12:5746-5754 (SCI)(Impact factor: 6.488;Rank: 16/141, 11.35%) 17. Wang HW, Hsueh CT, Lin CF, Chou TY, Hsu WH, Wang LS, Wu YC. Clinical implications of microsomal prostaglandin e synthase-1 overexpression in human non-small-cell lung cancer. Annals of Surgical Oncology (2006) 13(9):1224- 1234 (SCI)(Impact factor: 3.898;Rank: 9/148, 6.08%) 18. Yang MH, Chiang WC, Chou TY, Chang SY, Chen PM, Teng SC, Wu KJ. Increased NBS1 expression is a marker of aggressive head and neck cancer and overexpression of NBS1 contributes to transformation. Clinical Cancer Research (2006) 12(2):507-515 (SCI)(Impact factor: 6.488;Rank: 16/141, 11.35%) 19. Chou TY, Chiu CH, Li LH, Hsiao CY, Tzen CY, Chang KT, Chen YM, Perng RP, Tsai SF, Tsai CM. Mutation in the tyrosine kinase domain of epidermal growth factor receptor is a predictive and prognostic factor for Gefitinib treatment in patients with non-small cell lung cancer. Clinical Cancer Research (2005) 11(10):3750-3757 (SCI)(Impact factor: 6.488;Rank: 16/141, 11.35%) 20. Pan CC, Chen PC, Chou TY, Chiang H. Expression of calretinin and other mesothelioma-related markers in thymic carcinoma and thymoma. Human Pathology (2003) 34(11):1155-1162 (SCI)(Impact factor: 3.322;Rank: 17/68, 25.00%) 21. Small GW, Chou TY, Dang CV, Orlowski RZ. Evidence for involvement of calpain in c-Myc proteolysis in vivo. Archives of Biochemistry and Biophysics (2002) 400(2):151-161 (SCI)(Impact factor: 2.626;Rank: 30/70, 42.86%) 22. Chang CF, Su WJ, Chou TY, Perng RP. Hepatocellular carcinoma with Pancoast's syndrome as an initial symptom: a case report. Japanese Journal of Clinical Oncology (2001) 31(3):119-121 (SCI)(Impact factor: 1.405;Rank: 118/141, 83.69%) 23. Chou TY,* Hart GW. O-linked N-acetylglucosamine and cancer: messages from the glycosylation of c-Myc. Advances in experimental medicine and biology (2001) 491:413-418 (* Corresponding Author) 24. Huang YT, Lee TY, Lin HC, Chou TY, Yang YY, Hong CY. Hemodynamic 8 10 9. effects of Salvia miltiorrhiza on cirrhotic rats. Canadian Journal of Physiology and Pharmacology (2001) 79(7):566-572 (SCI)(Impact factor: 1.763;Rank: 45/74, 60.81%) 25. Hsu LH, Mao TL, Shih CS, Chou TY, Huang SN. Well-differentiated fetal adenocarcinoma of the lung. Journal of the Formosan Medical Association (2000) 99(5):425-7 (SCI)(Impact factor: 0.640;Rank: 83/107, 77.57%) 26. Liu YY, Lee YC, Chou TY, Perng RP. Pulmonary alveolar-septal amyloidosis presenting as heart failure with unilateral pleural effusion: a case report. Zhonghua Yi Xue Za Zhi (Taipei) (1999) 62(11):838-843 27. Sherwood JB, Carlson JA, Gold MA, Chou TY, Isacson C, Talerman A. Squamous metaplasia of the endometrium associated with HPV 6 and 11. Gynecologic oncology (1997) 66(1):141-145 (SCI)(Impact factor: 2.919;Rank: 12/61, 19.67%) 28. Jin YT, Tsai ST, Li C, Chang KC, Yan JJ, Chao WY, Eng HL, Chou TY, Wu TC, Su IJ. Prevalence of human papillomavirus in middle ear carcinoma associated with chronic otitis media. American journal of pathology (1997) 150(4):1327- 1333 (SCI)(Impact factor: 5.697;Rank: 2/68, 2.94%) 29. Li XM, Chopra RK, Chou TY, Schofield BH, Wills-Karp M, Huang SK. Mucosal IFN-gamma gene transfer inhibits pulmonary allergic responses in mice. Journal of immunology (1996) 157(8):3216-3219 (SCI)(Impact factor: 6.000;Rank: 16/121, 13.22%) 30. Chou TY, Hart GW, Dang CV. c-Myc is glycosylated at threonine 58, a known phosphorylation site and a mutational hot spot in lymphomas. Journal of biological chemistry (1995) 270(32):18961-18965 (SCI)(Impact factor: 5.520;Rank: 41/276, 14.86%) 31. Hoang AT, Lutterbach B, Lewis BC, Yano T, Chou TY, Barrett JF, Raffeld M, Hann SR, Dang CV. A link between increased transforming activity of lymphoma-derived MYC mutant alleles, their defective regulation by p107, and altered phosphorylation of the c-Myc transactivation domain. Molecular and cellular biology (1995)15(8):4031-4042 (SCI)(Impact factor: 5.942;Rank: 38/276, 13.77%) 32. Chou TY, Dang CV, Hart GW. Glycosylation of the c-Myc transactivation domain. Proceedings of the National Academy of Sciences of the United States of America (1995) 92(10):4417-4421 (SCI)(Impact factor: 9.380;Rank: 3/42, 7.14%) 33. Hart GW, Greis KD, Dong LY, Blomberg MA, Chou TY, Jiang MS, Roquemore EP, Snow DM, Kreppel LK, Cole RN, et al. O-linked N-acetylglucosamine: the "yin-yang" of Ser/Thr phosphorylation? Nuclear and cytoplasmic glycosylation. 9 10 10. AdvAdvances in experimental medicine and biology (1995) 376:115-123 34. Roquemore EP, Chou TY, Hart GW. Detection of O-linked N-acetylglucosamine (O-GlcNAc) on cytoplasmic and nuclear proteins. Methods in enzymology (1994) 230:443-460 (SCI)(Impact factor: 2.312;Rank: 33/65, 50.77%) 35. Haltiwanger RS, Kelly WG, Roquemore EP, Blomberg MA, Dong LY, Kreppel L, Chou TY, Hart GW. Glycosylation of nuclear and cytoplasmic proteins is ubiquitous and dynamic. Biochemical Society transactions (1992) 20(2):264- 269 (SCI)(Impact factor: 2.979;Rank: 117/276, 42.39%) 36. Chou TY,* Chen WY, Ho DM. Clear cell sarcoma of tendons and aponeuroses (malignant melanoma of soft parts): report of 2 cases. Taiwan Yi Xue Hui Za Zhi. (1989) 88(9):926-930 (* Corresponding Author) 10 10