Scientific Report 2010 – 2011

Aichi Cancer Center Research Institute

Chikusa-ku, Nagoya 464-8681 Japan (The Cover) Looking at the Aichi Cancer Center Research Institute Main Building over the drooping cherries in bloom.

Published by Dr. Kazuo Tajima Director Aichi Cancer Center Research Institute 1-1 Kanokoden, Chikusa-ku, Nagoya 464-8681, Japan Telephone: 052-762-6111 Facsimile: 052-763-5233

Editorial Committee Dr. Tatsuya Tsurumi (Division of Virology) Dr. Takayuki Murata (Division of Virology) Dr. Hiroshi Kumimoto (Division of Central Laboratory & Radiation Biology) Dr. Malcolm A. Moore, English Editor Ms. Hiromi Tamaki (Director's Office)

Printed by Nagoya University COOP 1 Furoucho, Chikusa-ku, Nagoya 464-0814, Japan Contents

Preface Kazuo Tajima 1 Organization of the Aichi Cancer Center Research Institute 2 SCIENTIFIC REPORTS Division of Epidemiology and Prevention General summary 5 1. Population-based Cancer Survival in Selected Registries in East Asia Ito, H., Matsuo, K., Tanaka, Hid., and Study group for Cancer Epidemiology and Statistics in East Asia. 6 2. The Hospital-based Epidemiologic Research Program at Aichi Cancer Center (HERPACC) Study Matsuo, K., Ito, H., Hosono, S., Kawase, T., Oze, I., Kawakita, D., Nakao, M., Hirose, K., Suzuki T., Chihara, D., Sueta, A., Sato, F., Islam, T., Sawamura, M., Yatabe, Y., Nakanishi T., Hasegawa, Y., Shinoda, M., Mitsudomi, T., Yamao, K., Iwata, H., Morishima Y., Hida, T., Seto, M., Murakami, S., Iida, S., Sato, S., Ueda, R., Nakamura, S., Iwase H., Tajima, K., and Tanaka, Hid. 7 2.1. Association between a GWAS-identified loci and risk of lung cancer in Japanese Ito, H., McKay, JD., Hosono, S., Hida, T., Yatabe, Y., Mitsudomi, T., Brennan, P., Tanaka, Hid., and Matsuo, K. 7 2.2. Weight gain during adulthood and body weight at age 20 are associated with the risk of endometrial cancer in Japanese women Hosono, S., Matsuo, K., Hirose, K., Ito, H., Suzuki T., Kawase, T., Watanabe, M., Nakanishi T., Tajima, K., and Tanaka, Hid. 8 2.3. Self-reported facial flushing after alcohol consumption and ALDH2 Glu504Lys polymorphism as risk factors for upper aerodigestive tract cancer in a Japanese population Oze, I., Matsuo, K., Hosono, S., Ito, H., Kawase, T., Watanabe, M., Suzuki, T., Hatooka, S., Yatabe, Y., Hasegawa, Y., Shinoda, M., Tajima, K., and Tanaka, Hid. 8 2.4. Association between dietary folate intake and clinical outcome in head and neck squamous cell carcinoma Kawakita, D., Matsuo, K., Sato, F., Oze, I., Hosono, S., Ito, H., Watanabe, M., Yatabe, Y., Hanai, N., Hasegawa, Y., Tajima, K., Murakami, S., and Tanaka, Hid. 9 2.5. ABO blood group alleles and the risk of pancreatic cancer in Japanese Nakao, M., Matsuo, K., Hosono, S., Ogata, S., Ito, H., Watanabe, M., Mizuno, N., Iida, S., Sato, S., Yatabe Y., Yamao, K., Ueda, R., Tajima, K., and Tanaka, Hid. 10 2.6. Inverse association between soy intake and non-Hodgkin lymphoma risk: a case-control study in Japan Chihara, D., Matsuo, K., Kanda, J., Hosono, S., Ito, H., Nakamura, S., Seto, M., Morishima Y., Tajima, K., and Tanaka, Hid. 10 2.7. Genetic risk prediction for breast cancer using a combination of low-

i penetrance polymorphisms in a Japanese population Sueta, A., Ito, H., Kawase, T., Hirose, K., Hosono, S., Yatabe, Y., Tajima, K., Tanaka, Hid., Iwata, H., Iwase H., and Matsuo, K. 11

Division of Oncological Pathology General summary 14 1. Molecular delivery systems using novel cell-penetrating peptides (CPPs) for anti-cancer medicine Saito, K., Matsushita, M.,and Kondo, E. 15 2. Potent anti-tumor killing activity of the multifunctional Treg cell line HOZOT against human tumors with diverse origins Inoue, T., Tashiro, Y., Takeuchi, M., Otani, T., Tsuji-Takayama, K., Okochi, A., Mukae, Y., Koreishi, M., Yamasaki, F., Kumon, H., Nakamura, S., Kibata, M. and Kondo, E. 16 3. miR-155, a modulator of FOXO3a protein expression, is underexpressed and is not upregulated by stimulation with HOZOT, a multifunctional Treg line Yamamoto, M., Kondo, E., Takeuchi, M., Harashima, A., Otani, T., Tsuji-Takayama, K., Yamasaki, F., Kumon, H., Kibata, M., and Nakamura, S. 17 4. Acquisition of an EMT phenotype by gefitinib-resistant cells of a head and neck squamous cell carcinoma cell line through Akt/GSK-3β/snail signaling Maseki, S., Ijichi, K., Tanaka, Ha., Fujii, M., Hasegawa, Y., Murakami. S, Kondo, E., and Nakanishi, H. 17 5. New multimodality imaging of gastric cancer peritoneal metastasis combining fluorescence imaging with near-infraredfluorophore-labeled antibody and MRI in mice Tsuchida, D., Matsushima, S., Ogasawara, M., Ito, A., Misawa, K., Ito, Y., Kondo, E., Kaneda , N., and Nakanishi, H. 17

Division of Molecular Oncology General summary 20 1. LATS2 is a tumor suppressor gene for malignant mesothelioma Murakami, H., Mizuno, T., Taniguchi, T., Fujii, M., Ishiguro, F., Fukui, T., Akatsuka, S., Horio, Y., Hida, T., Kondo, Y., Toyokuni, S., Osada, H., and Sekido, Y. 21 2. miR-375 is activated by ASH1 and inhibits YAP1 in a lineage-dependent manner in lung cancer Nishikawa, E., Osada, H., Okazaki Y., Arima, C., Tomida, S., Tatematsu, Y., Taguchi A., Shimada, Y., Yanagisawa, Y., Yatabe, Y., Toyokuni, S., Sekido, Y., and Takahashi, Ta. 21 3. Aberrant DNA methylation associated with aggressiveness of gastrointestinal stromal tumor Okamoto, Y., Sawaki, A., Ito, S., Nishida, T., Takahashi, T., Toyota, M., Suzuki, H., Shinomura, Y., Takeuchi, I., Shinjo, K., An, B., Ito, H., Yamao, K., Fujii, M., Murakami, H., Osada, H., Kataoka, H., Joh ,T., Sekido, Y., and Kondo, Y. 22

ii Division of Molecular Medicine General summary 25 1. Identification of FOXO3 and PRDM1 as tumor-suppressor gene candidates in NK-cell neoplasms by genomic and functional analyses Karube, K., Nakagawa, M., Tsuzuki, S., Takeuchi, I., Honma, K., Nakashima, Y., Shimizu, N., Ko, Y.H., Morishima, Y., Oshima, K., Nakamura, S., and Seto, M. 25 2. Synergistic effects of Bcl2, Myc and Ccnd1 in transforming mouse primary B cells into malignant cells Nakagawa, M., Honma, K., Tsuzuki, S., and Seto, M. 26 3. Establishment of an in vitro assay system allowing functional analysis of oncogenes in mature B cells Honma, K., and Seto, M. 26 4. Clonal evolution of adult T-cell leukemia/lymphoma takes place in lymph nodes Umino, A., Nakagawa, M., Utsunomiya, A., Tsukasaki, K., Taira, N., Katayama, N., and Seto, M. 27 5. Promotion and maintenance of leukemia by ERG Tsuzuki, S., and Seto, M. 27 6. Expansion of functionally defined mouse hematopoietic stem and progenitor cells by a short isoform of RUNX1/AML1 Tsuzuki, S., and Seto, M. 27

Division of Immunology General summary 29 1. Induction of cancer-specific CTL using artificial antigen presenting cells expressing an extrinsic HLA of interest while silencing intrinsic counterparts Demachi-Okamura, A., Kondo, S., Nishizawa, M., Yamamoto, E., Shibata, K., Kikkawa, F., and Kuzushima, K. 30 2. Zoledronate sensitizes ‘tumor-initiating cells’ from neuroblastoma lines to lysis by human gammadelta T cells Nishio, No., Tanaka, Y., Zhang, R., Hirosawa, T., Demachi-Okamura, A., Uemura, Y., Fujita, M., Taguchi, O., Takahashi, Y., Kojima, S., and Kuzushima, K. 30 3. Development of a novel adoptive immunotherapy using invariant natural killer T cells expressing a tumor-reactive T cell receptor Uemura, Y., Mineno, J., Ikeda, H., Shiku, H., and Kuzushima, K. 30 4. Mismatched human leukocyte antigen class II-restricted CD8⁺ cytotoxic T cells may mediate selective graft-versus-leukemia effects following allogeneic hematopoietic cell transplantation Hirosawa, T., Torikai, H., Yanagisawa, M., Kamei, M., Imahashi, N., Demachi-Okamura, A., Tanimoto, M., Shiraishi, K., Ito M., Miyamura, K., Shibata, K., Kikkawa, F., Morishima, Y., Takahashi, To., Emi, N., Kuzushima, K., and Akatsuka, Y. 31 5. Identification of minor histocompatibility antigens responsible for pulmonary graft-versus-host disease following adoptive immunotherapy Akatsuka, Y., Warren, E.H., Fujii, N., Brown, M.L., Ogawa, S., Matsubara, A., and Riddell, S.R. 32 6. Role of type 1 IFNs in antiglioma immunosurveillance - using mouse

iii studies to guide examination of novel prognostic markers in humans Fujita, M., Scheurer, M.E., Decker, S.A., McDonald, H.A., Kohanbash, G., Kastenhuber, E.R., Kato, H., Bondy, M.L., Ohlfest, J.R., and Okada, H. 32 7. COX-2 blockade suppresses gliomagenesis by inhibiting accumulation of myeloid-derived suppressor cells Fujita, M., Kohanbash, G., Fellows-Mayle, W., Hamilton, R.L., Komohara, Y., Decker, S.A., Ohlfest, J.R., and Okada, H. 33

Division of Virology General summary 36 1. Identification and characterization of CCAAT enhancer-binding protein (C/EBP) as a transcriptional activator for Epstein-Barr virus oncogene latent membrane protein 1 Noda, C., Murata, T., Kanda, T. and Tsurumi, T 36 2. Involvement of Jun dimerization protein 2 (JDP2) in the maintenance of Epstein-Barr virus latency Murata, T. and Tsurumi, T. 37 3. Transcriptional repression by sumoylation of Epstein-Barr virus BZLF1 protein correlates with histone deacetylase association Murata, T. and Tsurumi, T. 37 4. Tetrameric ring formation of Epstein-Barr virus polymerase processivity factor is crucial for viral replication Nakayama, S. and Tsurumi, T. 38 5. Spatiotemporally different DNA repair systems participate in Epstein-Barr virus genome maturation Sugimoto, A. and Tsurumi, T. 38 6. Unexpected instability of Family of Repeats (FR), the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems Kanda, T. and Tsurumi, T. 39 7. Human cytomegalovirus gene products essential for late viral gene expression assemble into prereplication complexes before viral DNA replication Isomura, H. and Tsurumi, T. 40

Division of Molecular Pathology General summary 42 1. Induction of selectin-ligand glycans during the epithelial-mesenchymal transition of colon cancer Sakuma, K., Aoki, M. and Kannagi, R. 42 2. Induction of sialyl Lewis x by epigenetic silencing of DTDST expression Yusa, A., Miyazaki, K., Lim, K.T., Yin, J., Yagi, H. and Kannagi, R. 43 3. Roles of the JNK/mTORC1 signaling in intestinal tumorigenesis in Apc mutant mice Aoki, M., Fujishita, T. and Taketo, MM. 44

Division of Biochemistry General summary 46 1. Novel regulation of Checkpoint kinase 1 (Chk1) Goto, H., Kasahara, K., Enomoto, M., Matsuyama, M., Li, P., iv Tomono, Y., Kiyono, T. and Inagaki, M. 46 2. Novel mitotic signaling from Akt/PKB to Polo-like kinase 1 Kasahara, K., Goto, H., Izawa, I., Kawamoto E., Watanabe, N., Tomono, Y., Kiyono, T. and Inagaki, M. 47 3. Functional analyses of vimentin phosphorylation in mice Matsuyama, M., Tanaka Hir., Kobori K., Izawa I., and Inagaki M. 48 4. Trichoplein blocks aberrant primary cilia assembly in proliferating cells Inoko, A., Matsuyama, M., Goto, H., Hayashi, Y., Ibi, M., He, D., Izawa, I., Urano, T., Yonemura, S., Kiyono, T. and Inagaki, M. 48

Central Service Unit General summary 51 1. Mitochondria DNA polymorphism Kumimoto, H. 51

Librarians Shibata, T., Yasuda, T., Sakou, M., and Horayama, K. 52

Publications 1 Journals 53 2 Reviews and books 72 3. Abstracts for international conferences 74

Records of seminars 80 Records of symposia 82 Author index for research reports and publications 88

v From left to right Ms. H. Tamaki, Dr, K. Tajima, and Ms. K. Mizutani. Preface ______

It is my great pleasure to share with you the 22nd Scientific Report (2010-2011) of the Aichi Cancer Center Research Institute. Since its establishment in 1965, Scientific Reports have been published biennially to document major research activities and highlight progress in and contributions to cancer research worldwide. As illustrated in the following pages, the Research Institute consists of 8 Divisions, the Central Service Unit, the Animal Facility and the Laboratory of Translational Research, focusing overall on three areas: cancer prevention/epidemiology; preclinical/experimental therapy; and carcinogenesis/molecular biology. In addition, the Research Institute is affiliated with the Nagoya University Graduate School of Medicine and the Nagoya City University Graduate School of Pharmaceutical Sciences and at the present time 7 professors and 4 associate professors nominated from our chiefs and section heads are engaged in education of graduate school students from the two universities. A total of 76 staff members, comprising 42 researchers and 18 research assistants, as well as 16 research residents, are now conducting a wide range of studies, together with 7 graduate school students, 32 visiting research fellows and approximately 40 temporary research assistants.

The major areas being pursued are as follows: - descriptive and analytical epidemiology of cancers - primary and secondary prevention of cancer - molecular pathogenesis of intractable malignancies - peptide-based anti-cancer DDS technology - molecular oncology of respiratory tract cancer - molecular biology of translocation-junction genes of hematopoietic tumors - basic studies for cancer immunotherapy - oncogenicity, molecular biology and immunology of DNA tumor viruses - glycobiology of cancer cells in relation to metastasis - genetic dissection of signaling pathways in colon cancer - molecular mechanisms of cell proliferation and movement - involvement of repair mechanisms in carcinogenesis

More detailed descriptions of the research topics of each Division are provided in the body of the report. It is our sincere hope that the activities of the Institute will make major contributions to elucidation of the mechanisms of carcinogenesis and to development of novel clinical applications in cancer diagnosis, treatment and prevention. Finally, I would like to express my deep appreciation to the Aichi Prefectural Government for the continuous support received since this Institute was founded in 1964. Granting support from the Ministry of Education, Science, Sports, Culture and Technology, the Ministry of Health, Labor, and Welfare, and the Ministry of Economy, Trade and Industry, Japan, in addition to other related organizations, is also gratefully acknowledged.

January, 2012

Kazuo Tajima, M.D., M.P.H., D.M.Sci. Director

1 Organization of the Aichi Cancer Center Research Institute ______

2 SCIENTIFIC REPORTS From left to right First row: Dr. I. Oze, Dr. H. Ito, Dr. K. Matsuo, Dr. H. Tanaka, Dr. S. Hosono, Ms. M. Watanabe Second row: Dr. A. Sueta, Ms. T. Ito, Ms. Y. Mano, Ms. S. Inui, Ms. K. Fukaya, Ms. Y. Yama- uchi, Ms. A. Uematsu, Ms. A. Yoshida, Ms. H. Tanaka Third row: Dr. D. Chihara, Dr. M. Nakao, Ms. S. Torii, Ms. T. Nishiwaki, Ms. S. Irikura, Ms. R. Saito, Ms. M. Nakahama, Ms. M. Yamaguchi, Ms. Y. Hamajima Fourth row: Dr. D. Kawakita, Dr. M. Sawamura, Ms. Y. Matsubara, Ms. Y. Tanaka, Ms. M. Ishida, Ms. K. Saito, Ms. A. Hiraiwa, Ms. I. Kato Inset: Dr. Y. Morishima, Dr. T. Islam, Dr. K. Fukumoto, Dr. M. Higashi, Ms. K. Suganuma, Ms. C. Taniguchi, Ms. T. Sanami, Ms. M. Matsubara, Ms. M. Miyoshi

4 Division of Epidemiology and Prevention ______

Hideo Tanaka, M.D., PhD. Chief Keitaro Matsuo, M.D., PhD., S.M. Section Head (as of April 2008) Hidemi Ito, M.D., PhD., M.P.H. Section Head (as of April 2009) Takakazu Kawase, M.D., PhD. Senior Researcher (until March 2010) Satoyo Hosono, M.D., PhD. Senior Researcher (as of April 2009) Isao Oze, M.D. Research Resident (until March 2010) Daisuke Kawakita, M.D. Research Resident (as of April 2010) Makoto Nakao, M.D. Research Resident (as of April 2010) Miki Watanabe, Research Assistant (as of April 2006) Misako Nakahama, Research Assistant (as of April 2009) Yasuo Morishima, M.D. PhD. Researcher (as of April 2011)

Visiting Trainees Kiyonori Kuriki, B.P., D.M.Sc. University of Shizuoka Hideo Nakane, PhD. Toyota Central R&D Labs., Inc. Isao Oze, M.D. Research Resident of Foundation for Promotion of Cancer Research and Okayama Univer- sity Graduate School of Medicine Masao Nishira, M.D. Nagoya University Graduate School of Medicine Tania Islam, M.D. Nagoya University Graduate School of Medicine Dai Chihara, M.D. Nagoya University Graduate School of Medicine Aiko Sueta, M.D. Kumamoto University Graduate School of Medicine Makiko Higashi, M.D. Nagoya University Graduate School of Medicine Koichi Fukumoto, M.D. Nagoya University Graduate School of Medicine Chi-young Park, M.D. Nagoya University Graduate School of Medicine Toshiki Okasaka, M.D. Nagoya University Graduate School of Medicine Junya Kanda, M.D. Kyoto University Graduate School of Medicine Daisuke Ennishi, M.D. Okayama University Graduate School of Medicine Fumihito Sato, D.D., Nagoya University Graduate School of Medicine Masatsugu Sawamura, D.D., Nagoya University Graduate School of Medicine Takiko Tateishi, M.S. FALCO biosystems Ltd. Tokai central laboratory

General Summary The current research activities of the Division of Epidemiology and Prevention cover the following six subjects: (1) descriptive epidemiology of cancer incidence, mortality and survival using data from the Aichi Prefectural Cancer Registry and other population-based registries in collaborative studies; (2) analytical epidemiology based on the hospital-based epidemiologic research program at Aichi Cancer Center (HERPACC) to determine risk and protective factors for cancer, with a particular focus on gene-environmental interactions; (3) clinical epidemiology for elucidating optimal cancer treatment and prevention; (4) genome-wide associations to detect polymorphisms associated with cancer risk; and (5) anti-tobacco policy and smoking cessation therapy. Our main results in 2010 and 2011 were that; 1) international differences in population-based cancer survival are considerable in East Asian countries; 2) the15q25 region modifies the effect of smoking on the risk of lung cancer in Japanese; 3) weight gain during adulthood is associated with increased risk of endometrial cancer in Japanese women; 4) facial flushing is less useful to predict upper aerodigestive tract (UAT) cancer risk than genotyping the ALDH2 polymorphism; 5) pre-treatment high dietary folate intake is an independent prognostic factor in head and neck squamous cell carcinoma (HNSCC) patients; 6) ABO blood group alleles are significantly associated with pancreatic cancer risk in Japanese; 7) an inverse association between soy intake and non-Hodgkin lymphoma risk exists; and 8) we could establish a method of genetic risk prediction for breast cancer using a combination of low-penetrance polymorphisms in our Japanese population.

5 1. Population-based Cancer Survival in Manila and Rizal, the Philippines, Registries in Selected Registries in East Asia Cixian and Qidong, China, and the Taiwan Cancer Ito, H., Matsuo, K., Tanaka, Hid., and Study group for Registry joined in this study. The participating reg- Cancer Epidemiology and Statistics in East Asia istries submitted data on five-year relative survival Reliable population-based cancer survival data rates of the stomach, colon, rectum, liver, lung, are essential for assessment of the effectiveness of breast and uterine cervix, diagnosed from 2000 to cancer screening programs, determine the most ap- 2002. All registries satisfied the criteria for data propriate cancer therapy and provide details on quality: a percentage of death certificate only cases prevalent cancer cases. International comparisons <20% and a percentage of completeness of fol- are useful to allow societies, mass media and health low-up at 5 years after diagnosis of <5%. authorities to gain a real appreciation of the cancer Figure 1 illustrates the remarkable results ob- program in their own country and provide an impe- tained. Compared to the registries in Korea, China tus to improve registration and cancer control plan- and Philippines, all five Japanese registries showed ning. Since directly comparable survival data substantially higher five-year relative survival rates among Asian countries are presently very limited, (5-RSR) for stomach cancer (64~71% in the Japa- we performed cancer survival estimates in East nese registries; 19~53% in others) and lung cancer Asia within the framework of the 2nd cooperative (20~28% for males and 34~47% for females in the study on population-based cancer survival in East Japanese registries; 13~7% for males and 8~21% in Asia in 2010. the others). Most of the Japanese registries showed The Korean Central Cancer Registry (KCCR), higher 5-RSR for colorectal cancer than the others five Cancer Registries in Japan (Miyagi, Yama- (59~79% in the Japanese registries; 12~64% in the gata, Niigata, Fukui and Nagasaki), Registries in others). The RSRs for colorectal cancer in males

Fig. 1. Five-year relative survival rate in each cancer site (2000-2002). (A) Stomach cancer (male), (B) Lung cancer (male), (C) Female Breast cancer, (D), Cervical Cancer.

6 and females in Korea (64% and 61%) were close to *6 Department of Thoracic Surgery, Aichi Cancer Center those in the Japanese registries. Whereas most of Hospital. *7 the 5-RSRs for liver cancer in the Japanese regis- Department of Gastroenterology, Aichi Cancer Center Hospital. tries were between 21% and 29%, those in the oth- *8 ers were also relatively low (2~22%). However, the Department of Breast Oncology, Aichi Cancer Center Hospital. 5-RSR for liver cancer in males and females in *9 Department of hematology and Cell therapy, Aichi Taiwan (20% and 22%) were close to those in the Cancer Center Hospital. Japanese registries. In each registry, the 5-RSR for *10 Department of Thoracic Oncology, Aichi Cancer Cen- female breast cancer showed the highest figure ter Hospital. among all cancer sites. The Japanese and Taiwan *11 Division of Molecular Medicine, Aichi Cancer Center registries and the KCCR showed values of more Research Institute. *12 than 80%, whereas the registries in Manila & Rizal Department of Otorhinolaryngology, Head and Neck and China showed relatively low rates ranged from Surgery, Nagoya City University Graduate School of Medical Sciences. 37% to 72%. On the other hand, the 5-RSR for cer- *13 vical cancer in the KCCR and Taiwan registry were Department of Pathology and Clinical Laboratories, Nagoya University Hosopital. highest (81% and 76%, respectively), followed by *14 Department of Breast and Endocrine Surgery, Ku- the values in the Japanese registries, ranging from mamoto University Graduate School of Medical Science. 70% to 78%. Manila & Rizal (52%), Cixian (27%) and Qidong (22%) all showed relatively low 5-RSR 2.1 Association between a GWAS-identified for cervical cancer. loci and risk of lung cancer in Japanese This study is unique as the first attempt to cal- Ito, H., McKay, JD.*1, Hosono, S., Hida, T.*2, Yatabe, culate population-based cancer survival in selected Y.*3, Mitsudomi, T.*4, Brennan, P.*5, Tanaka, Hid., and registries in East Asia with a focus data quality. Our Matsuo, K. results showed substantial variation in the reported Background: Genome-wide association studies RSRs among countries in the early 2000s. These (GWAS) have identified 15q25 and 5p15 as lung differences apparently reflect differences in cancer cancer risk chromosomal regions in Caucasians. registration systems as well as in cancer control ac- The genetic structures of these loci differ between tivities and cancer care. Therefore, we need to con- Asians and Caucasians, however, indicating a need sider all of these factors in interpretation of results. for additional studies in Asian populations. To ex- amine the impact of 15p25 and 5p15 on lung cancer 2. The Hospital-based Epidemiologic risk and smoking intensity, we conducted a Research Program at Aichi Cancer case-control study in Japanese. We also focused on Center (HERPACC) Study whether these loci modify the effect of smoking Matsuo, K., Ito, H., Hosono, S., Kawase, T., Oze, I., behavior on lung cancer risk. *1 *2 Kawakita, D., Nakao, M., Hirose, K. , Suzuki T. , Methods: The subjects were 716 Japanese pa- Chihara, D., Sueta, A., Sato, F., Islam, T., Sawamura, M., tients with lung cancer and 716 controls. Associa- *3 *4 *5 Yatabe, Y. , Nakanishi T. , Hasegawa, Y. , Shinoda, tions were examined by logistic regression models *6 *6 *7 *8 M. , Mitsudomi, T. , Yamao, K. , Iwata, H. , with adjustment for potential confounders. Morishima Y.*9, Hida, T.*10, Seto, M.*11, Murakami, S.*12, Results: We found that the variants of Iida, S.*2, Sato, S.*2, Ueda, R.* 2, Nakamura, S.*13, Iwase rs12914385 and rs931794 on 15q25 modified the H.*14, Tajima, K., and Tanaka, Hid. effect of cumulative tobacco smoking on lung cancer risk, but that these two loci showed no statisti- *1 Department of Planning and Information, Aichi Pre- cally significant main influences on lung cancer risk. fectural Institute of Public Health. Compared to never-smoking without the risk allele *2 Department of Medical Oncology and Immunology, of rs931794, the OR for heavy-smoking without the Nagoya City University. *3 Department of Pathology and Molecular Diagnostics, risk allele was 4.03 (95%CI, 2.45-6.62) and that Aichi Cancer Center Hospital. with the risk allele was 8.09 (5.09-12.9), and the *4 Department of Gynecologic Oncology, Aichi Cancer joint effect of rs931734 and cumulative tobacco Center Hospital. consumption was statistically significant *5 Department of Head and Neck Surgery, Aichi Cancer (p-interaction <0.001). A similar impact was ob- Center Hospital. served with rs12914385 on chromosome 15q25

7 (p-interaction=0.021). Associations for the OR = 2.02 (95%CI = 1.38-2.96); p-trend <0.001]. TERT-CLPM1L locus on 5p15 with lung cancer Parity and BMI at age 20 appeared to modify the risk in Japanese were of a similar magnitude to effect of weight gain on endometrial cancer risk, those in Caucasians. albeit without statistical significance. This positive Conclusions: These results support contributions association of weight gain on risk was observed of 15q25 and 5p15 to lung cancer and indicate that only for endometrioid adenocarcinoma. the 15q25 region modifies the well-established ef- These results suggest the presence of a positive fect of smoking on the risk of lung cancer in our association between weight gain during adulthood Japanese population. and endometrial cancer risk among Japanese women. Further investigations of these findings are *1 Genetic Cancer Susceptibility Group, International warranted. Agency for Research on Cancer. *2 Department of Thoracic Oncology, Aichi Cancer Cen- *1 Department of Planning and Information, Aichi Pre- ter Hospital. *3 fectural Institute of Public Health. Department of Pathology and Molecular Diagnostics, *2 Department of Medical Oncology and Immunology, Aichi Cancer Center Hospital. *4 Nagoya City University. Department of Thoracic Surgery, Aichi Cancer Center *3 Department of Gynecologic Oncology, Aichi Cancer Hospital. Center Hospital. *5 Genetic Epidemiology Group, International Agency for Research on Cancer. 2.3. Self-reported facial flushing after alcohol consumption and ALDH2 2.2. Weight gain during adulthood and body Glu504Lys polymorphism as risk weight at age 20 are associated with factors for upper aerodigestive tract the risk of endometrial cancer in cancer in a Japanese population Japanese women Oze, I., Matsuo, K., Hosono, S., Ito, H., Kawase, T., Hosono, S., Matsuo, K., Hirose, K.*1, Ito, H., Suzuki T.*2, Watanabe, M., Suzuki, T., Hatooka, S.*1, Yatabe, Y.*2, Kawase, T., Watanabe, M., Nakanishi T.*3, Tajima, K., Hasegawa, Y.*3, Shinoda, M.*1, Tajima, K., and Tanaka, and Tanaka, Hid. Hid. Although current obesity is an established risk Some Japanese exhibit facial flushing after factor for endometrial cancer, the role of weight drinking alcohol, this being considered due to acet- gain during adulthood and obesity in early adult- aldehydemia. The concentration of blood acetalde- hood on endometrial cancer has not been elucidated. hyde is direcly related with the catalytic activity of Here, we conducted a case-control study with 222 acetaldehyde dehydrogenase (ALDH) and an histologically diagnosed incident endometrial can- ALDH2 polymorphism (rs671, Glu504Lys) is cer cases and 2,162 age- and menstrual-status known to be associated with upper aerodigestive matched non-cancer controls. tract (UAT) cancer due to its effects on enzyme ac- Information on current body weight, weight and tivity. It remains controversial whether facial height at age 20 years, and lifestyle/environmental flushing is useful in predicting UAT cancer risk as a factors was obtained from a self-administered ques- surrogate marker of ALDH2 polymorphism. There- tionnaire. Subjects were classified into three groups fore, we conducted the present case-control study according to change in body mass index (BMI, with 961 UAT cancer patients and 2883 matched kg/m2) from that at age 20 years to enrollment non-cancer outpatients of Aichi Cancer Center (BMI change ≤ 0 as reference, BMI change 0-3, and Hospital. Information on facial flushing and other BMI change ≥ 3). Impact of adult BMI change or lifestyle factors was collected via self-administered obesity in early adulthood was evaluated using an questionnaire. Associations between facial flushing, unconditional logistic regression model adjusted for the polymorphism and UAT cancer were assessed potential confounders. with reference to odds ratios and 95% confidence Higher BMI at age 20 (BMI ≥ 25) showed a sig- intervals using conditional logistic regression mod- nificant positive association with endometrial can- els. Facial flushing had no significant association cer risk (P = 0.005), as did BMI increase during with UAT cancer, although the ALDH2 Lys allele adulthood [0-3 BMI change, multivariate odds ratio was significantly linked to increased risk. No sig- (OR) = 1.28 (95%CI = 0.88-1.87); > 3 BMI change, nificant interaction between facial flushing and

was made for alcohol status, smoking status and early adulthood weight in the analysis as possible confounders. Soy intake was significantly associated with a reduced risk of NHL in women but not in men (OR [95%CI] for moderate and high intake: women, 0.64 [0.42-1.00] and 0.66 [0.42-1.02], respectively; men, 1.40 [0.87-2.24] and 1.33 [0.82-2.15], respectively; p-interaction = 0.02). This finding appeared consistent across NHL subtypes. No reproductive factors, including age of menarche, menopausal status, parity and age of first delivery, were found to interact with soy intake. These Fig. 3. Receiver-operating-characteristic-curves (ROC) in the risk models. results indicate the potential The straight dotted line with an AUC of 50% is the reference. The AUC of tential importance of certain in- the upper black curved line, which represents the genetic risk model in gredients in soy for lymphoma- addition to the established risk factors, is 69.33%, whereas that of the genesis. Further studies to gray line representing the established risk model only is 66.52% (P = evaluate possible underlying 1.3×10-4), and that of the lowest curved line representing the genetic risk mechanisms are warranted. model only is 59.65%. *1 Department of Pathology and that bind competitively to estrogen receptors but Clinical Laboratories, Nagoya University Hosopital. have a weak estrogenic effect nearly 0.1% of that of *2 Division of Molecular Medicine, Aichi Cancer Center estradiol. Although soy intake has been associated Research Institute. with a reduced risk of several cancers such as breast, *3 Department of hematology and Cell therapy, Aichi prostate, colorectal, and gastric cancer, its signifi- Cancer Center Hospital. cance with NHL is not known. We therefore evaluated whether soy intake 2.7. Genetic risk prediction for breast might impact on risk of NHL by conducting a hos- cancer using a combination of pital-based case-control study in 302 cancer pa- low-penetrance polymorphisms in a tients and 1510 age- and sex-matched controls. In Japanese population accordance with our a priori hypothesis that the ef- Sueta, A., Ito, H., Kawase, T., Hirose, K.*1, Hosono, S., fect of soy on NHL risk may differ by sex, as it Yatabe, Y.*2, Tajima, K., Tanaka, Hid., Iwata, H.*3, does for other types of cancer, sex-stratified analy- Iwase H.*4, and Matsuo, K. sis for evaluation of effect modification was defined Genome-wide association studies (GWASs) as the default model. Analysis after stratification by have identified genetic variants associated with reproductive factors was also performed. We also breast cancer. However, most conducted to date evaluated the impact of soy on the three histological have focused on women of European descent, and subtypes, namely diffuse large B-cell lymphoma, the contributions of variants as predictors in Japa- follicular lymphoma and marginal zone B-cell nese women have remained unclear. Here, we ana- lymphoma.. Odds ratios (OR) and 95% confidence lyzed 23 genetic variants identified in previous intervals (CI) for groups with moderate (27-51 GWASs and conducted a case-control study with g/day) to high (> 51 g/day) relative to low (< 27 697 case subjects and 1394 age- and menopausal g/day) intake were calculated using a multivariate status-matched controls. conditional logistic regression model. Adjustment Associations between variants and breast cancer

11 risk were assessed by odds ratios (ORs) and 95% 3.01 (1.97 - 4.58) and 8.69 (2.75 - 27.5), respec- -9 confidence intervals (CIs) using conditional logistic tively (P trend =1.9×10 ). The c statistic for a model regression models, adjusted for age, age at men- including the genetic risk score in addition to the arche, menopausal status, current body-mass-index conventional risk factors was 0.6933, versus 0.6652 (BMI), age at first live birth, regular exercise, fa- with conventional risk factors only (P = 1.3× 10-4) mily history of breast cancer, and referral pattern to (Figure 1). The population-attributable fraction of our hospital. In addition, we created a polygenetic the risk score was 33.0%. risk score, using those variants with a statistically In conclusion, we could establish a model for significant association with breast cancer risk, and genetic risk prediction for breast cancer in a Japa- also evaluated the contribution of these genetic pre- nese population. This approach featuring a genetic dictors using the c statistic. risk score may prove useful in distinguishing We observed significant associations between women at high risk of breast cancer from those at eleven single nucleotide polymorphisms (SNPs) low risk, particularly in the context of targeted and breast cancer risk. A dose-dependent associa- prevention. tion was observed between the risk of breast cancer *1 and the genetic risk score, which was an aggregate Department of Planning and Information, Aichi Pre- fectural Institute of Public Health. measure of alleles in seven selected variants, *2 namely FGFR2-rs2981579, Department of Pathology and Molecular Diagnostics, TOX3/TNRC9-rs3803662, C6orf97-rs2046210, Aichi Cancer Center Hospital. *3 Department of Breast Oncology, Aichi Cancer Center 8q24-rs13281615, SLC4A7-rs4973768, Hospital. LSP1-rs38137198 and CASP8-rs10931936. Com- *4 Department of Breast and Endocrine Surgery, Kuma- pared to women with scores of 3 or less, ORs for moto University Graduate School of Medical Science. women with scores of 4-5, 6-7, 8-9 and 10 or more were 1.33 (95% CI, 1.00 - 1.80), 1.71 (1.26 - 2.30),

12 From left to right First row: Dr. Susumu Nakata, Dr. Eisaku Kondo, Dr. Hayao Nakanishi Second row: Dr. Takuya Saito, Ms. Misao Niwa, Ms. Yukina Ichikawa, Ms. Kikuko Hanaoka, Ms. Noriko Saito Third row: Ms. Mayumi Yoshimura, Dr. Ken Saito, Mr. Akihiro Ito

13 Division of Oncological Pathology ______

Eisaku Kondo, M.D.,Chief Hayao Nakanishi, M.D., Section Head Susumu Nakata, M.D., Senior Researcher (as of April 2011) Ken Saito, Ph.D., Research Resident (as of August 2010) Tomoko Mori, Ph.D., Research Resident (until Oct 2011) Eriko Koya, Ph.D., Research Resident (until Jan 2011) Makoto Matsui, Ph.D., Research Resident (until March 2010) Noriko Saito, Research Assistant Keiko Nishida, Semi-regular Employee Junya Yamamoto, Semi-regular Employee (as of Jan 2012) Yuko Saito, Semi-regular Employee (as of Jan 2012) Misao Niwa, Semi-regular Employee (Until Dec 2011) Sayaka Ohsiden, Semi-regular Employee (Until March 2010) Takashi Isomura, Semi-regular Employee (Until March 2010)

Visiting Trainees Mitsuhiko Ota, D.D.S., School of Dentistry, Aichi-Gakuin University Yukiko Oshima, M.D., Dept. of Surgery II, Nagoya University School of Medicine Hiroki Murakami, M.D., Dept. of Surgery II, Nagoya University School of Medicine Maseki Shinichiro, M.D., Dept. of Otorhinolaryngology, Nagoya City University School of Medicine Takuya Saito, M.D., Dept. of Surgery, Pref. Osaka Saiseikai Izuo Hospital Yuki Tomita, M.D., Dept. of Medical Oncology and Immunology, Nagoya City University School of Medicine Daisuke Tsuchida, M.S., Graduate School of Pharmaceutical Sciences, Meijo University Mai Ogasawara, Graduate School of Pharmaceutical Sciences, Meijo University Akihiro Ito, Graduate School of Pharmaceutical Sciences, Meijo University

General Summary The aim for our research at the Division of Oncological Pathology is to disclose the pathogenesis of human malignancies of diverse origins through molecular analyses based on data obtained by morphological and biological examination of cancer tissues and human cancer cells. Our interest is now mainly focused on acquiring novel pathological findings of human intractable malignancies, including tumors of digestive, respiratory and hematopoietic organs as a basic research strategy, and on development of advanced medical technology for application in the clinic. In our basic pathological research, we are working on identification of key molecules contributing to metastasis and invasion, paying particular attention to morphological and biological characterization of cancer stem cells. Since November 2009, as the major current research subject, we are also pioneering new fields in trials to develop novel peptide-based molecular diagnostic and therapeutic systems (DDS) utilizing current information based on the medical studies. Another important responsibility of our division is to conduct autopsies. Postmortem examinations give us valuable information on the behavior of neoplasms and their response to therapy, assessing the effectiveness or failure of current therapies to clarify pathogenesis in cancer patients. Thus, our present overall aim is to promote comprehensive pathological research which can make real contributions to current tumor medicine.

14 1. Molecular delivery systems using novel carcinoma cells. These tumor-lineage-homing CPPs cell-penetrating peptides (CPPs) for were more efficiently uptaken into target cells in in anti-cancer medicine vitro and in vivo assays as compared with TAT and Saito, K., Matsushita, M.*1, and Kondo, E. other conventional CPPs. Our hope is that our The development of molecular delivery systems unique tumor-homing CPPs may prove useful for based on cell-penetrating peptides (CPPs) has re- peptide-based non-invasive medical technology cently attracted much attention because of non-invasive properties in vivo. Our interest is in developing novel tumor-lineage-homing CPPs that are highly permeable to cancer cells according to their tumor origins. A specific random peptide library was employed for isolating CPPs encoded by novel artificial amino acid sequences differing from conventional CPPs such as TAT, pAnt (antennapedia), and HIV gp41. Screening of over 40 of these CPPs, revealed 10 exhibiting specific responses that were efficiently incorporated into tumor cells of particular origins. For example, CPP2 highly penetrated cells derived from colon adenocarcinomas, while CPP44 targeted human myeloid leukemia cells as well as hepatocellular

Figure. In vivo anti-tumor activity of HOZOT in the WiDr-bearing NOD/SCID mouse model. Eradica- tion of disseminated WiDr cells was observed in HOZOT-treated mice (right panels) but not in non-treated animals (left panels) on examination of the peritoneal cavity, lung and liver.

relationship between EMT and resistance to epi- 3. miR-155, a modulator of FOXO3a protein dermal growth factor receptor (EGFR) targeting expression, is underexpressed and is drugs in head and neck squamous cell carcinomas not upregulated by stimulation with (HNSCC) remains unknown. In this study, we in- HOZOT, a multifunctional Treg line vestigated acquisition of EMT by an HNSCC cell Yamamoto, M.*1, Kondo, E., Takeuchi, M.*1, Harashima, line (UMSCC81B). We isolated a fibroblastoid A.*1, Otani, T.*1, Tsuji-Takayama, K.*1, Yamasaki, F.*2, variant (81B-Fb) from gefitinib-resistant Kumon, H.*3, Kibata, M.*1, and Nakamura, S.*1 UMSCC81B-GR3 cells obtained after increasing MicroRNAs (miRNAs) play important roles in doses of gefitinib treatment in vitro and examined regulating post-transcriptional gene repression in a EMT and its underlying mechanism. 81B-Fb cells variety of immunological processes. In particular, exhibied fibroblast-like morphology, increased mo- much attention has been focused on their roles in tility, loss of E-cadherin, and acquisition of regulatory T (Treg) cells which are crucial for vimentin and snail expression. In 81B-Fb cells, maintaining peripheral tolerance and controlling T down-regulation of EGFR, mediated by increased cell responses. Recently, we established a novel ubiquitination, and activation of downstream prote- type of human Treg cell line, termed HOZOT, fea- in kinase B (Akt), glycogen synthase kinase-beta turing multifunctional cells exhibiting a (GSK-3) signaling and up-regulation of snail ex- CD4+CD8+ phenotype. In this study, we performed pression were observed as compared with parental miRNA profiling to identify signature miRNAs of UMSCC81B cells. LY294002, but not U0126, HOZOT, and therein identified miR-155. Although suppressed fetal bovine serum or heregulin miR-155 has also been characterized as a signature 1-induced phosphorylation of Akt/GSK-3 and miRNA for FOXP3+ natural Treg (nTreg) cells, it snail expression, while inhibiting 81B-Fb cell mo- was expressed quite differently in the HOZOT case. tility. Furthermore, enforced expression of EGFR Under both stimulatory and non-stimulatory condi- resulted in partial restoration of gefitinib-sensitivity tions, miR-155 expression remained at low levels in and reversal of EMT. These results suggest that HOZOT, while its expression in nTreg and conven- EMT in gefitinib-resistant cells is mediated by tional T cells remarkably increased after stimulation. down-regulation of EGFR and compensatory acti- We next searched for candidate target genes of vation of the Akt/GSK-3/snail pathway. miR-155 through bioinformatics, and identified

FOXO3a, a negative regulator of Akt signaling, as a *1 miR-155 target gene. Further studies by gain- and Department of Otolaryngology-Head & Neck Surgery, loss-of-function experiments supported a role for Nagoya City University Graduate School of Medical Sci- ences miR-155 in the regulation of FOXO3a protein ex- *2 Department of Head & Neck Surgery,Aichi Cancer pression in conventional T and HOZOT cells. Center Central Hospital

*1 Cell Biology Institute, Research Center, Hayashibara 5. New multimodality imaging of gastric Biochemical Laboratories Inc., *2 Kurashiki Medical Center cancer peritoneal metastasis *3 Department of Urology, Okayama University Graduate combining fluorescence imaging with School of Medicine, Dentistry and Pharmaceutical Sci- near-infraredfluorophore-labeled ences antibody and MRI in mice Tsuchida, D.*1, Matsushima, S.*2, Ogasawara, M.*1, Ito, 4. Acquisition of an EMT phenotype by A.*1, Misawa, K.*3, Ito, Y.*3, Kondo, E., Kaneda , N.*1, gefitinib-resistant cells of a head and and Nakanishi, H. neck squamous cell carcinoma cell line Peritoneal metastasis is the most frequent pattern through Akt/GSK-3β/snail signaling of recurrence after curative surgery for gastric can- Maseki, S.*1, Ijichi, K.*1, Tanaka, Ha., Fujii, M., cer. However, such recurrence is difficult to detect Hasegawa, Y.*2, Murakami. S*1, Kondo, E., and by conventional computed tomography (CT) and Nakanishi, H. magnetic resonance imaging (MRI) at early stages. The epithelial mesenchymal transition (EMT) is To improve the sensitivity and specificity of diag- known to be associated with chemoresistance as nostic imaging for peritoneal metastasis, we have well as increased invasion/metastasis. However, the developed a new type of multimodality approach

17 specific optical imaging was obtained using an NIR fluorophore-labeled anti-human CEA antibody instead of cetuximab. Although whole-body fluorescence imaging had higher sensitivity for detection of small-sized peritoneal metastases (1-2 mm in diameter) than MRI, it proved difficult to accurately determine organ distribution of the metastasis. Thus, our newly developed multimodality imaging system with fusion of 3D optical and MRI images combining fluorescence with near-infrared (NIR) demonstrated improved diag- fluorophore-labeled antibodies and MRI. Dual op- improved diagnostic accuracy over either method tical imaging using luciferase-tagged gastric cancer alone. The present results suggest that multimodal- cell lines with different EGFR expression and ity imaging with NIR fluorophore-labeled EGFR or XenoLight CF750 or indocyanine-green CEA antibody in combination with MRI should al- (ICG)-labeled anti-human EGFR antibody (cetuxi- low sensitive, specific and anatomically accurate mab) allowed successful visualization of metastatic detection of peritoneal metastasis at an early stage. foci in the peritoneal cavity noninvasively even *1 with low EGFR expressing tumors. This fluores- Graduate School of Pharmaceutical Sciences, Meijo University cence imaging proved to be specific, as evidenced *2 by the fact that fluorescence signaling from metas- Department of Diagnostic and Interventional Radiol- tases was abolished by blocking with an excess ogy,Aichi Cancer Center Central Hospital *3Department of Gastroenterological Surgery, Aichi amount of unlabeled antibody. Similar sensitive and Cancer Center Central Hospital, Nagoya, Japan

18 From left to right First row: Ms. Yasue Matsudaira, Ms. Mika Yamamoto, Dr. Yoshitaka Sekido, Dr. Keiko Shinjo Second row: Dr. Ichidai Tanaka, Mr. Yoshio Tatematsu, Dr. Hirotaka Osada, Dr. Momen El- shazley, Mr. Keisuke Katsushima, Dr. Futoshi Ishiguro, Dr. Yutaka Kondo, Dr. Fumiharu Ohka Inset: Dr. Makiko Fujii

19 Division of Molecular Oncology ______

Yoshitaka Sekido M.D., Ph.D., Chief Hirotaka Osada, M.D., Ph.D., Section Head Yutaka Kondo, M.D., Ph.D., Section Head Hideki Murakami, M.D., Ph.D., Senior Researcher (until March 2011) Makiko Fujii, D.D.S., PhD., Senior Researcher Yoshio Tatematsu, B.S., Research Assistant Yasue Matsudaira, B.S., Research Assistant Ikuko Tomimatsu, Semi-regular Employee (until October 2011) Mika Yamamoto, Semi-regular Employee (as of November 2009) Kimiko Tanaka, Semi-regular Employee (as of September 2011)

Research Resident Tetsuya Mizuno, M.D., Nagoya University School of Medicine (until September 2011) Futoshi Ishiguro, M.D., Nagoya University School of Medicine

Visiting Trainees Keiko Shinjo, M.D., Nagoya University School of Medicine Yasuyuki Okamoto, M.D., Nagoya City University School of Medicine Yugo Kishida M.D., Nagoya University School of Medicine Haixing Ju, M.D., Zhejiang Cancer Hospital Keisuke Katsushima, M.S., Hokkaido University Graduate School of Life Science Daiki Ito, B.S., Nagoya University School of Medicine Fumiharu Ohka, M.D., Nagoya University School of Medicine Ichidai Tanaka, M.D., Nagoya University School of Medicine Asuki Fukatsu, M.D., Nagoya University School of Medicine Momen Elshazley, M.D., Sohag University, Egypt Hidemi Ito, Phar.B., Meijo University Mayuko Tuji, Meijo University Ryosuke Kondo, Meijo University

General Summary Our goal is to determine genetic lesions and epigenetic alterations giving rise to human solid cancers and use this information for their prevention, diagnosis and treatment. Currently, we are focusing on lung cancer, malignant mesotheliomas, colon cancer, hepatomas, gastrointestinal mesenchymal tumors (GIST) and brain tumors. In particular we are making use of opportunities to dissect biochemical and pathological pathways underlying malignant phenotypes including dysregulated cell proliferation and differentiation, invasion and metastasis. Human cancers arise because of genetic mutations in oncogenes and tumor suppressor genes and our approach is to study candidate genes, applying systematic molecular analysis of biochemical pathways, global microarrays for gene expression profiling and comparative genomic hybridization techniques for chromosomal abnormalities. Epigenetic changes like DNA methylation, histone modification, and micro- RNA expression also allow clues not only to inactivation of tumor suppressor genes but also to fundamental mechanisms for maintenance of cancer stem cell populations and differentiated cell lineages of each tissue. We also functionally analyze candidate genes by transducing wild type copies into human cancer cells and testing for their ability to suppress malignancy in vitro and in vivo as well as characterizing their protein products biochemically. Alternatively, we inactivate their expression using RNA interference (RNAi) in either tumor or normal cells and then dissect the resulting phenotypes. Understanding the functions of mutated genes and disrupted signaling pathways will provide foundations for a translational research approach to human malignancies, from bench to bedside.

20 1. LATS2 is a tumor suppressor gene data indicate that Hippo pathway dysregula- for malignant mesothelioma tion is frequent, with inactivation of LATS2 Murakami, H., Mizuno, T., Taniguchi, T.*1, Fujii, or an upstream regulator of this pathway, M., Ishiguro, F., Fukui, T.*2, Akatsuka, S.*3, Merlin, which is encoded by NF2. Thus, our Horio, Y.*4, Hida, T.*4, Kondo, Y., Toyokuni, S.*3, results suggest that the inactivation of Osada, H., and Sekido, Y. LATS2 is one of the key mechanisms for Malignant mesothelioma (MM) is an ag- constitutive activation of YAP, which in- gressive neoplasm associated with asbestos duces deregulation of MM cell proliferation. exposure. To identify new MM cancer-related genes we carried out ge- 1Department of Cardio-Thoracic Surgery, Na- nome-wide array-based comparative genomic goya University Graduate School of Medicine, hybridization analysis using 14 MM cell lines Nagoya, Japan that we established in our laboratory. Three 2Department of Thoracic Surgery, Aichi Cancer cell lines showed overlapping homozygous Center Hospital, Nagoya, Japan deletion at chromosome 13q12, harboring the 3Department of Pathology and Biological Large tumor suppressor homolog 2 (LATS2) Responses, Graduate School of Medicine, gene. With 6 other MM cell lines and 25 MM Nagoya University, Nagoya, Japan tumors, we found 10 inactivating homozygous deletions or mutations of LATS2 among 4Department of Thoracic Oncology, Aichi Cancer 45 MMs. LATS2 encodes a serine/threonine Center Hospital, Nagoya, Japan kinase, a component of the Hippo tumor suppressive signaling pathway, and we trans- 2. miR-375 is activated by ASH1 and duced LATS2 in MM cells featuring mutation. inhibits YAP1 in a Transduction of LATS2 inactivated oncopro- lineage-dependent manner in lung tein YAP, a transcriptional co-activator, via cancer phosphorylation and inhibited MM cell pro- Nishikawa, E.*1, Osada, H., Okazaki Y.*2, Arima, liferation. Additional immunohistochemical C.*1, Tomida, S.*1, Tatematsu, Y., Taguchi A.*1, analysis showed 13 of 45 MM tumors to ex- Shimada, Y.*1, Yanagisawa, Y.*3, Yatabe, Y.*4, hibit low expression of LATS2. Since NF2 is Toyokuni, S.*2, Sekido, Y., and Takahashi, Ta.*1 genetically mutated in 40-50% of MM, our

Fig. 1. Lineage-dependent growth-regulating effects of YAP1 in lung cancers. A, time course of expression of each miR-375 predicted target gene after Pre-miR-375 transfection. Among the miR-375 predicted target genes, transcriptional coactivator YAP1 was the most significantly repressed after miR-375 transfection. B, flow cytometric analysis of lentivirus-infected A549 and ACC-LC- 172 fluorescence-positive cells. Cells were infected with YAP1- expressing or empty lentiviruses carrying the fluorescent protein Venus, and analyzed at 3 and 9 days after infection. The fluorescence-positive population was moderately increased in the A549 cells infected with the YAP1-virus, whereas it almost disappeared in similarly infected ACC-LC-172 cells.

21 Lung cancers with neuroendocrine (NE) to establish epigenetic profiles associated features are often very aggressive but the with the malignant transformation of GISTs. underlying molecular mechanisms remain Methylation of four tumor suppressor genes, elusive. The transcription factor RASSF1A, p16, CDH1, and MGMT was ASH1/ASCL1 is a master regulator of pul- analyzed in GISTs. Additionally, genome- monary NE cell development that is suspect- wide DNA methylation profiles were com- ed to be involved in the pathogenesis of lung pared between small, malignant-prone, and cancers with NE features (NE-lung cancers). malignant GISTs using methylated GpG is- Here we report definition of microRNA land amplification microarrays (MCAM) in a miR-375 as a key downstream effector of training set (n=40). Relationships between ASH1 function in NE-lung cancer cells. the methylation status of genes identified by miR-375 was thus markedly induced by MCAM and clinical features of the disease ASH1 in lung cancer cells where it proved were tested in a validation set (n=75). sufficient to induce NE differentiation. Methylation of RASSF1A progressively in- miR-375 upregulation was a prerequisite for creased from small to malignant GISTs. p16 ASH1-mediated induction of NE features. was specifically methylated in malig- The transcriptional coactivator YAP1 was nant-prone and malignant GISTs. MCAM determined to be a direct target of miR-375. analysis showed that more genes were meth- Furthermore, YAP1 showed a negative cor- ylated in advanced than in small GISTs (av- relation with miR-375 in a panel of lung can- erage of 473 genes vs 360 genes, respectively, cer cell lines and growth inhibitory activity in P=0.012). Interestingly, the methylation pro- NE-lung cancer cells. Our results have eluci- file of malignant GISTs was prominently af- dated an ASH1 effector axis in NE-lung fected by their location. Two genes, REC8 cancers that is functionally pivotal in con- and PAX3, which were newly-identified via trolling NE features and the alleviation from MCAM analysis, were differentially YAP1-mediated growth inhibition. methylated in small and malignant GISTs in the training and validation sets. Patients with *1 Division of Molecular Carcinogenesis, Nagoya methylation of at least REC8, PAX3, or p16 University Graduate School of Medicine, Nagoya, had a significantly poorer prognosis Japan (P=0.034). Our results suggest that GIST is *2Department of Pathology and Biological Re- not, in epigenetic terms, a uniform disease sponses, Graduate School of Medicine, Nagoya University, Nagoya, Japan and that DNA methylation in a set of genes is *3Institute for Advanced Research, Nagoya Uni- associated with aggressive clinical behavior versity, Nagoya, Japan and unfavorable prognosis. The genes identi- *4Department of Pathology and Molecular Diag- fied may potentially serve as biomarkers for nostics, Aichi Cancer Center Hospital predicting aggressive GISTs with poor sur- vivability. 3. Aberrant DNA methylation *1 associated with aggressiveness Department of Gastroenterology, Aichi Cancer of gastrointestinal stromal tumor Center Hospital *2Department of Gastroenterological surgery, Ai- Okamoto, Y., Sawaki, A.*1, Ito, S.*2, Nishida, T.*3, *4 *5 *6 chi Cancer Center Hospital Takahashi, T. , Toyota, M. , Suzuki, H. , *3Department of Surgery, Osaka Police Hospital, *6 *7 Shinomura, Y. , Takeuchi, I. , Shinjo, K., An, B., Osaka, Japan Ito, H.*8, Yamao, K.*1, Fujii, M., Murakami, H., *4Department of Surgery, Osaka General Medical Osada, H., Kataoka, H.*9, Joh ,T.*9, Sekido, Y., Center, Osaka, Japan *5 and Kondo, Y.*10 Department of Biochemistry, Sapporo Medical University, Hokkaido, Japan The majority of gastrointestinal stromal *6 tumors (GISTs) have KIT mutations; how- First Department of Internal Medicine, Sapporo ever, epigenetic abnormalities that could Medical University, Hokkaido, Japan *7Department of Computer Science, Nagoya In- conceivably potentiate the aggressiveness of stitute of Technology, Nagoya, Japan GISTs are largely unidentified. Our aim was *8Division of Epidemiology and Prevention, Aichi

22 Cancer Center Research Institute *10Precursory Research for Embryonic Science *9Department of Gastroenterology and Metabo- and Technology (PRESTO), Japan Science and lism, Nagoya City University Graduate School of Technology Agency Medical Sciences, Nagoya, Japan

Fig. 2. Methylated GpG island amplification microarray (MCAM) analysis in GISTs. A, Box-and-whisker plots of the number of methylated genes in small GISTs, as well as advanced GISTs (malignant-prone and malignant GISTs). The mean is marked by a horizontal line inside the box whose ends denote the upper and lower quartiles. Error bars represent 5- and 95-percentile values. *, P=0.012. B, Heat-map overview of 766 genes, which are commonly methylated in more than 30% of the cases in either small GISTs, malignant-prone GISTs or malignant GISTs (left). Venn diagram of this set of 766 genes showing the relationship of methylation target genes in the three classified groups (right). A number of genes were specifically methylated in advanced GISTs; malignant-prone GISTs (172 genes) and malignant GISTs (155 genes, highlighted in blue). C, Principal Components Analysis of the 155 genes associated with gastric GISTs (left) and small intestinal GISTs (middle), along with merged image of gastric malignant GISTs and small intestinal malignant GISTs (right). The first three principal components accounted for 81.7% of the total variance. Ellipsoids of standard deviations (75% prob- ability) are drawn with respect to each group. Magenta, small GISTs; deep green, malignant-prone GISTs from stomach; green, malignant GISTs from stomach; orange, malignant-prone GISTs from small intestine; yellow, malignant GISTs from small intestine.

23 From left to right First row: Ms. Y. Kasugai, Dr. K. Karube, Dr. M. Seto, Dr. S. Tsuzuki Second row: Dr. K. Yamamoto, Dr. L. Fang, Dr. H. Kato, Ms. K. Hirano, Dr. N. Yoshida, Ms. S. Sato, Dr. K. Arita

24 Division of Molecular Medicine ______

Masao Seto, M.D., Ph.D. Chief Shinobu Tsuzuki, M.D., Ph.D. Section Chief Masao Nakagawa, M.D., Ph.D. Senior Researcher (until June 2011) Kennosuke Karube, M.D., Ph.D. Senior Researcher Keiichiro Honma, M.D., Ph.D. Senior Researcher (until June 2011) Yumiko Kasugai, B.S. Research Assistant Kiyoko Yamamoto, M.D. Research Resident Liu Fang, M.D. Research Resident Akira Umino, M.D. Research Resident (until March 2011) Kyoko Hirano, B.S. Research Assistant

Visiting Trainees Harumi Kato, M.D. Nagoya University Graduate School of Medicine Kotaro Arita, M.D. Toyama University Graduate School of Medicine Noriaki Yoshida, M.D. Hiroshima University School of Medicine

General Summary Research in this laboratory is focused on elucidating genetic and molecular bases of human cancer, with a view to applying the obtained knowledge to clinical oncology. We are currently working on hematological malignancies, in collaboration with physicians in the Department of Hematology and Cell Therapy (Chief, Dr. Tomohiro Kinoshita) of Aichi Cancer Center Central Hospital. Hematological malignancies are highly associated with genetic changes so that some hematological malignancies can be classified according to genetic changes specific to given disease entities. Such close association provides evidence that the genetic changes play pivotal roles in disease development and/or clinicopathological manifestations. Over the last two years, we have studied several issues in particular: (1) identification of candidate tumor suppressor genes residing in the common region of deletion in chromosome 6q in natural killer cell lymphomas; (2) co-operativity of Bcl2, Myc and Ccnd1 in transforming immature B lymphocytes; (3) establishment of a novel culture system allowing functional screening of genes involved in genesis of malignant lymphomas; (4) clonal evolution of adult T cell leukemias/lymphomas with special reference to lymph nodes as sites of occurrence; (5) functional roles of ERG, a transcription factor whose high expression is associated with poor prognosis in T cell acute lymphoblastic leukemia and cytogenetically normal acute myelogenous leukemia; and (6) ability of a short isoform of a transcription factor AML1/RUNX1 to effect expansion of hematopoietic stem/progenitor cells.

1. Identification of FOXO3 and PRDM1 as other included LACE1 and FOXO3 (see Figure). tumor-suppressor gene candidates in All genes located in these regions, except for NK-cell neoplasms by genomic and POPDC3 and AIM1, were down-regulated in neo- functional analyses plastic samples, as determined by gene-expression Karube, K., Nakagawa, M., Tsuzuki, S., Takeuchi, I.*1, analysis, and were therefore considered to be can- Honma, K., Nakashima, Y.*2, Shimizu, N.*3, Ko, Y.H.*4, didate tumor-suppressor genes. A20 and HACE1, Morishima, Y.*5, Oshima, K.*6, Nakamura, S.*7, and Seto, the well-known tumor-suppressor genes located on M. 6q21-23, were included as candidates because they Oligo-array comparative genomic hybridization also demonstrated frequent genomic deletion and (CGH) and gene-expression profiling of natural down-regulated expression. The Tet-Off NK cell killer (NK)-cell neoplasms were used in an effort to line NKL was subsequently established for func- delineate the underlying molecular pathogenesis. tional analyses. Seven candidate genes were trans- Oligo-array CGH identified two 6q21 regions that duced into Tet-Off NKL and forced re-expression were most frequently deleted (14 of 39 or 36% of was induced. Re-expression of FOXO3 and cases). One of these regions included POPDC3, PRDM1 suppressed NKL proliferation, but this was PREP, PRDM1, ATG5, and AIM1, whereas the not the case after re-expression of the other genes. This effect was confirmed using another NK cell

25 ture, colony formation and oncogenicity in vivo were assessed in mouse primary B cells exogenously expressing various combinations of Bcl2, Myc and Ccnd1. For the functional screening, Bcl2- and Myc-expressing primary B cells were infected with a retroviral cDNA library. Inserted cDNA of transformed cells in culture were then identified. RESULTS: Primary B cells exogenously expressing Bcl2, Myc and Ccnd1 showed factor-independent growth, an enhanced capacity for colony-formation and aggressive oncogenicity, unlike the cases observed with expression of any combination of only two of the genes. line, SNK10. Furthermore, genomic analyses de- We thus could identify CCND3 and NRAS as genes tected nonsense mutations of PRDM1 that led to cooperating with Bcl2 and Myc through the func- functional inactivation in one cell line and one tional screening. CONCLUSIONS: Bcl2, Myc and clinical sample. Our findings provide evidence that Ccnd1 or Bcl2, Myc and CCND3 synergistically PRDM1 and FOXO3 play important roles in the transform mouse primary B cells into aggressive pathogenesis of NK-cell neoplasms. malignant cells. Our new synergism-based method *1 Department of Computer Science/Scientific and En- should prove useful for the identification of other gineering Simulation, Nagoya Institute of Technol- synergistic gene combinations in tumor develop- ogy, Nagoya, Japan ment, and thus expand our systematic understand- *2 Department of Hematology, Iizuka Hospital, Iizuka, ing of a wide range of cancer-causing elements. Japan *3 Department of Virology, Division of Virology and 3. Establishment of an in vitro assay Immunology, Medical Research Institute, Tokyo system allowing functional analysis of Medical and Dental University, Tokyo, Japan oncogenes in mature B cells *4 Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Su- Honma, K., and Seto, M. won, Korea We have been searching for genomic alterations *5 Department of Hematology and Cell Therapy, Aichi in lymphoma clinical samples using an array CGH Cancer Center, Nagoya, Aichi, Japan method. Candidate oncogenes or oncosuppressor *6 Department of Pathology, School of Medicine, Ku- genes thus found need to be functionally validated rume University, Kurume, Japan to reveal their detailed roles in lymphoma devel- *7 Department of Pathology and Clinical Laboratories, opment. Although we have established a system Nagoya University Hospital, Nagoya, Japan using immature B cells as a target of genetic manipulation, an equivalent system using mature B 2. Synergistic effects of Bcl2, Myc and cells, normal counterparts of lymphoma cells, has Ccnd1 in transforming mouse primary hitherto been lacking. Here, we established a cul- B cells into malignant cells ture system that allows maturation of immature B to Nakagawa, M., Honma, K., Tsuzuki, S., and Seto, M. mature B cells in vitro, which is usable for func- BACKGROUND: Synergistic effects resulting tional analysis of candidate onco- or oncosupressor from combination of BCL2 and MYC or MYC and genes. Specifically, immature B cells derived from CCND1 have been implicated in genesis of human mouse fetal liver were cultivated on a layer of B-cell lymphomas. Although the identification of NIH-3T3 cells engineered to express CD40ligand, other cooperative genes involved is important, our Baff, and CXCL13, and then stimulated with present understanding of this area remains scant. anti-IgM antibodies in combination with lipopoly- The objective of this study was therefore to identify saccharides. By doing so, the immature B cells were any additional cooperative gene(s) associated with found to efficiently differentiate into IgG-positive BCL2 and MYC or MYC and CCND1. First, we mature B cells. Our preliminary use of this culture assessed whether Bcl2, Myc and Ccnd1 could co- system allowed identification of a truncated SIRT1 operate. Next, we developed a synergism-based gene, expressed in a lymphoma cell line, as a can- functional screening method for the identification of didate gene involved in constitutive activation of other oncogene(s) that act in concert with Bcl2 and HIF1alpha; HIF1 alpha is a known oncogene in Myc. DESIGN AND METHODS: Growth in cul- lymphoma development. Overall, the results sug-

26 gest that our in vitro system will provide a novel differentiation and induces expansion of T and tool for the analysis of functional roles of onco- erythroid cells and increases frequencies of myeloid genes or onco-suppressors in lymphomas. progenitors in mouse BM transplantation models. The expanded T cells then give rise to T-cell acute 4. Clonal evolution of adult T-cell lymphoblastic leukemia after acquisition of muta- leukemia/lymphoma takes place in tions in the Notch1 gene. Targeted expression of lymph nodes ERG in B cells also altered differentiation and Umino, A., Nakagawa, M., Utsunomiya, A.*8, Tsukasaki, promoted growth of precursor B cells. Overall, the- K.*9, Taira, N.*10, Katayama, N.*11, and Seto, M. se findings suggest a general role of ERG in pro- Adult T-cell leukemia/lymphoma (ATLL) is the moting growth of adult hematopoietic cells of vari- neoplasm caused by human T-cell leukemia virus type ous lineages. In line with this concept, 1 (HTLV-1). On performance of oligo-array compara- shRNA-mediated silencing of ERG expression at- tive genomic hybridization (CGH) with paired pe- tenuated growth of a variety of human leukemia cell ripheral blood (PB) and lymph node (LN) samples lines. Thus, ERG is capable of promoting the de- from 13 patients with acute ATLL, we found that the velopment of leukemia and appears crucial for dis- genome profiles frequently differed. Some 9 of 13 ease maintenance. cases investigated had a log2 ratio imbalance among chromosomes, and such imbalance was more frequent 6. Expansion of functionally defined mouse in LN specimens. Detailed analysis revealed that the hematopoietic stem and progenitor likely cause was presence of multiple subclones. Five cells by a short isoform of of the 13 cases showed homozygous loss of regions in RUNX1/AML1 PB samples, which were not found in their LN coun- Tsuzuki, S., and Seto, M. terparts, indicating that tumors in the PB were derived Self-renewal is essential for the maintenance from LN subclones in most cases. Southern blot and regeneration of the hematopoietic system. analysis of TCR showed that multiple subclones to However, the search for molecules capable of pro- have originated from common clones. We conclude moting self-renewal and expanding hematopoietic that in many ATLL cases, multiple subclones in LNs stem cells (HSCs) has hitherto met with limited originate from a common clone, and that particular LN success. Here we show that a short isoform subclones then appear in the PB. (AML1a) of RUNX1/AML1 has such activity. Thus enforced AML1a expression expanded functionally defined HSCs with an efficiency that was at least 20 *8 Department of Hematology, Imamura Bun-in Hospital, Kagoshima, Japan times greater than that of the control in vivo and by *9 Department of Hematology and Molecular Medicine 18-fold within 7 days ex vivo. The ex Unit, Atomic Bomb Disease Institute, Nagasaki University vivo-expanded HSCs could repopulate hosts fol- Graduate School of Biomedical Sciences, Nagasaki, Japan lowing secondary transplantations. Moreover, *10 Department of Internal Medicine, Heartlife Hospital, AML1a expression resulted in vigorous and Okinawa, Japan long-term (>106-fold at 4 weeks) ex vivo expansion *11 Hematology and Oncology, Mie University Gradu- of progenitor cell populations capable of ate School of Medicine, Tsu, Japan multi-lineage differentiation. Gene expression analysis revealed AML1a expression to be associated 5. Promotion and maintenance of leukemia with upregulation of many genes, including Hoxa9, by ERG Meis1, Stat1 and Ski. shRNA-mediated silencing of Tsuzuki, S., and Seto, M. these genes attenuated AML1a-mediated activities. The Ets-related gene (ERG) located on human Overall, these findings establish AML1a as an iso- chromosome 21 encodes a transcription factor form-specific molecule that can influence several thought to be causally related to Down syn- transcriptional regulators associated with HSCs, drome-associated acute megakaryocytic leukemia in leading to enhanced self-renewal activity and he- childhood. In clinical adult cases, while increased matopoietic stem/progenitor cell expansion ex vivo expression of ERG is indicative of poor prognosis and in vivo. The results may have implications for in T-cell acute lymphoblastic leukemia and cytoge- HSC transplantation and transfusion medicine, netically normal acute myeloid leukemia, an eti- given that effects can also be obtained by ological role remains elusive. Here, we show that cell-penetrating AML1a protein. enforced expression of ERG in adult BM cells alters

27 From left to right First row: Ms. M. Nishizawa, Dr. Y. Uemura, Dr. K. Kuzushima, Dr. M. Fujita, Dr. A. De- machi-Okamura Second row: Mr. H. Maki, Ms. K. Shiraishi, Dr. R. Zhang, Ms. T. Tsuboi, Ms. R. Miura, Dr. S. Kondo, Ms. M. Tatsumi Inset: Dr. Y. Akatsuka

28 Division of Immunology ______

Kiyotaka Kuzushima, M.D. Chief Mitsugu Fujita, M.D. Section Head (as of April 2011) Yasushi Uemura, D.D.S. Senior Researcher (as of April 2010) Nobuhiro Nishio, M.D. Senior Researcher (as of April 2010 until June 2011) Ayako Demachi-Okamura, Ph.D. Researcher Tomoya Hirosawa, M.D. Research Resident (until Dec 2010) Shinji Kondo, M.D. Research Resident (as of April 2010) Rong Zhang, Ph.D. Research Resident (as of May 2011) Miwako Nishizawa, B.P. Senior Research Assistant Jingwen Wang, Ph.D. Semi-regular Employee (until March 2010) Rieko Miura, Semi-regular Employee (as of July 2011) Minako Tatsumi, Semi-regular Employee (as of July 2011) Keiko Shiraishi, Semi-regular Employee Yasuko Watarai, Semi-regular Employee (until August 2010) Tomiko Tsuboi, Semi-regular Employee

Visiting Researcher Yoshiki Akatsuka, M.D. Department of Hematology & Oncology, Fujita Health University

Visiting Trainees Tomoya Hirosawa, M.D. Department of Gynecologic Oncology, Aichi Cancer Center (as of January 2011) Hiroyuki Maki, Nagoya University Graduate School of Medicine (as of March 2010)

General Summary The object of our research is to establish molecular and cellular bases for future establishment of novel cancer therapies applying immune responses. The achievements during past two years are here summarized. Firstly, we established a novel artificial antigen presenting cell (aAPC) system which could overcome the absence of autologous tumor cell lines for CTL stimulation. Briefly, it is a combination of:1) silencing endogenous HLA class I expression by means of a short interfering RNA (siRNA); and 2) transducing siRNA-resistant HLA-A24 cDNA harboring synonymous codons for the targeted sequence of siRNA into a cancer cell line. Resultant lines can express a single allele of HLA molecules that makes it eligible for aAPC to induce CTL restricted to that allele. In fact we have already established CTL lines and derivative clones from CD8+ T cells stimulated with an ovarian cancer cell line equipped with such HLA modifications. Now, identification of genes recognized by the CTL clones is on going by expression cloning methods applying a cDNA library made from mRNA of the cancer cell line. Secondly, we have focused on γδT cells and neuroblastoma, a childhood malignancy, and shown that zoledronate efficiently sensitizes tumor cells to lysis by ex vivo-expanded γδT cells. Importantly, outgrowth of neuroblastoma tumor-initiating cells was inhibited by the T cells, in both in-vitro and in-vivo experiments. We hope that the findings will facilitate development of anti-neuroblastoma immunotherapy using ex vivo-expanded γδT cells. Thirdly, we have established invariant natural killer T cell lines expressing TCR α/β genes specific to an MAGE A4 peptide presented by HLA-A24 molecules. The T cells could be shown to be reactive to the peptide and α-galactosylceramide presented by CD1d molecules. Their anti-tumor potential is currently being evaluated in vitro and in a mouse model. Fourthly, we have characterized an interesting CD8+ CTL clone restricted to HLA-DR molecules isolated from a patient following an HLA-DR-mismatched hematopoietic stem cell transplantation. The clone exerted anti-leukemia activity in mouse experiments. Precursor frequency analysis revealed a significant fraction of the total donor CTL responses towards the individual mismatched HLA-DR antigen in two patients, underscoring unexpectedly significant CTL responses in the context of HLA class II. In addition, a minor

29 histocompatibility antigen was identified with the aid of International HapMap Project B-lymphoid cell lines and their genetic data in collaboration of the Fred Hutchinson Cancer Research Center and the Graduate School of Medicine, University of Tokyo. Lastly, we have started to investigate de novo glioma tumorigenesis and immunological interventions using mice intracerebrally transfected with NRAS and a short hairpin RNA against P53.

1. Induction of cancer-specific CTL using *1 Department of Obstetrics and Gynecology, Nagoya artificial antigen presenting cells University Graduate School of Medicine expressing an extrinsic HLA of interest while silencing intrinsic counterparts 2. Zoledronate sensitizes ‘tumor-initiating Demachi-Okamura, A., Kondo, S., Nishizawa, M., cells’ from neuroblastoma lines to lysis Yamamoto, E. *1, Shibata, K. *1, Kikkawa, F. *1, and by human gammadelta T cells *1 Kuzushima, K. Nishio, No., Tanaka, Y. , Zhang, R., Hirosawa, T., Cytotoxic T lymphocytes (CTLs) exert Demachi-Okamura, A., Uemura, Y., Fujita, M., Taguchi, *2 *3 *3 anti-tumor effects through recognition of tumor an- O. , Takahashi, Y. , Kojima, S. , and Kuzushima, K. tigen-derived peptides bound to human leukocyte Neuroblastoma, the most common extracranial antigens (HLAs) on cell surfaces. Examples which solid tumor in children, is refractory to intensive can be used to probe for such peptides are important multimodal therapy. Some neuroblastoma cell lines, reagents to define tumor antigens. Generating tu- when cultured in serum-free medium with certain mor-specific CTLs usually requires autologous tu- growth factors, form spheres which contain tumor cell lines. However, we have successfully in- mor-initiating cells (TICs). They are believed to be duced cancer-specific CTLs using artificial antigen responsible for tumor formation, as well as resis- presenting cells (aAPCs) that have endogenous tu- tance to conventional therapy, so that an optimal mor-associated peptides on given HLA molecules strategy of tumor immunotherapy should target this but not their own HLAs. With this approach, aAPC minor population. has now been developed for application for any We have shown that zoledronate efficiently cells having their own HLAs. We have sought to sensitizes both neuroblastoma cells and such use aAPCs expressing a single HLA molecule pre- sphere-forming cells to lysis by ex vivo-expanded senting tumor-associated peptides. Vγ9Vδ2 T cells, a T cell population that recognizes TOV21G is an HLA-A24-negative cell line de- and kills tumor cells irrespective of their surface rived from an ovarian cancer. We first abolished in- HLA expression. Cytotoxic activity could be aug- trinsic HLA expression of TOV21G cells by trans- mented by addition of IL-15 and/or IL-18 during T fection of a short interfering RNA (siRNA) for the cell culture. Furthermore, outgrowth of neuroblas- HLA class I gene. We next lentivirally introduced a toma TICs was inhibited by co-culture with CD86 gene and siRNA-resistant HLA-A24 cDNA Vγ9Vδ2 T cells, as demonstrated by in-vitro colony having synonymous codons for the targeted se- formation assay and in an in-vivo mouse model. We quence of siRNA into TOV21G cells. Naïve CD8+ conclude that ex vivo-expanded Vγ9Vδ2 T cells are T lymphocytes of HLA-A24-positive donors were a promising tool for anti-neuroblastoma immuno- stimulated by the TOV21G-based aAPCs, and sev- therapy. eral CTL clones were obtained. Most produced IFN-γ against HLA-A24-expressing TOV21G cells, *1 Center for Innovation in Immunoregulative Technol- but not the parental TOV21G and ogy and Therapeutics, Graduate School of Medicine, HLA-A24-positive fibroblast cells. Some of clones Kyoto University *2 Division of Molecular Pathology, Aichi Cancer Center lysed not only HLA-A24-expressing TOV21G but Research Institute also other ovarian cancer cell lines. Currently, we *3 Department of Pediatrics, Nagoya University Graduate are attempting to identify antigens recognized by School of Medicine the CTL clones, employing a cDNA expression cloning method. 3. Development of a novel adoptive These results suggest that aAPC featuring immunotherapy using invariant natural siRNA-mediated HLA modification provides a killer T cells expressing a useful tool to establish CTL lines targeting a variety tumor-reactive T cell receptor of cancer cells with restriction to a desired HLA al- Uemura, Y., Mineno, J. *1, Ikeda, H. *2, Shiku, H. *2, and lele. Kuzushima, K.

30 Antigen-specific cancer immunotherapy is a *1Center for Cell and Gene Therapy, Takara Bio Inc. *2 promising strategy to achieve therapeutic responses. Department of Immuno-Gene Therapy, Mie University Currently, T cell epitope-peptide vaccines are under Graduate School of Medicine development by many different groups and clinical trials have already been performed. Unfortunately, 4. Mismatched human leukocyte antigen outcomes have been disappointing. It is conceivable class II-restricted CD8⁺ cytotoxic T cells that this is at least partly due to: 1) difficulty in ex- may mediate selective panding antigen-specific T cells in vivo; and 2) graft-versus-leukemia effects following immune suppressive effects of tumor microenvi- allogeneic hematopoietic cell ronments. transplantation Improvement of T cell-mediated immune re- Hirosawa, T.*1, Torikai, H. *2, Yanagisawa, M.*3, Kamei, sponses can hopefully be achieved by development M.*4, Imahashi, N.*3, Demachi-Okamura, A., Tanimoto, of a new adoptive immunotherapy in which T cells M., Shiraishi, K., Ito M.*5, Miyamura, K.*3, Shibata, K.*1, with an adjuvant property to overcome the immune Kikkawa, F.*1, Morishima, Y.*6, Takahashi, To.*7, Emi, suppressive network and an ability to kill cancer N.*8, Kuzushima, K., and Akatsuka, Y.*8 cells are expanded ex vivo and given back to the pa- Allogeneic hematopoietic stem cell transplan- tient. To address this possibility, we have focused tation (allo-HSCT) is established as one of the most on a unique subpopulation of T lymphocytes, effective therapeutic options for hematopoietic ma- namely, invariant natural killer T (iNKT) cells. lignancies. Its major benefits are obtained from the These are characterized by the expression of an in- allo-immunity directed against patients' leukemia variant T cell receptor (TCR) that recognizes cells known as the graft-versus-tumor (GVT) effect. α-galactosylceramide (α-GalCer) presented by However, the same allo-immune reactions can also CD1d on antigen-presenting cells. They comprise be directed against normal host tissues resulting in two major distinct subsets displaying either graft-versus-host disease (GVHD). Although HSCT CD4-CD8β- (double negative: DN) or CD4+CD8β- from human leukocyte antigen (HLA)-identical sib- (CD4). We previously reported that activated DN lings or unrelated donors is feasible to minimize the iNKT cells exerted cytotoxic activity while activat- risk of acute graft-versus-host disease (aGVHD), ed CD4 iNKT cells exhibited adjuvant properties by HSCT from HLA-mismatched donors can also be inducing IL-12p70 production in dendritic cells performed when a patient has advanced disease and (DCs). no HLA matched donor is available. In these cases, In the current study, CD8α/β and TCRα/β CD8+ or CD4+ T cell responses are induced de- genes obtained from an HLA-A24-restricted and pending on the mismatched HLA class I or II al- MAGE-A4-specific T cell clone were retrovirally lele(s). Herein, we describe an + transduced into iNKT cells. The resultant HLA-DRB1*08:03-restricted CD8 CTL clone, TCR-transduced iNKT cells showed cytotoxicity in named CTL-1H8, isolated from a patient following response to MAGE-A4 peptide-loaded T2-A24 an HLA-DR-mismatched HSCT from his brother. cells and tumor cells expressing both MAGE-A4 Lysis of a patient Epstein-Barr virus-transformed B and HLA-A24. The cytotoxic activity of the cell line (B-LCL) by CTL-1H8 was inhibited after TCR-transduced DN iNKT cells was comparable to the addition of blocking antibodies against that of TCR-transduced conventional CD8 T cells. HLA-DR and CD8, whereas antibodies against In contrast, although TCR-transduced CD4 iNKT pan-HLA class I or CD4 had no effect. The cells exhibited marginal cytotoxic activity, they 1H8-CTL clone did not lyse recipient dermal fibro- acted as a cellular adjuvant inducing robust blasts whose HLA-DRB1*08:03 expression was IL-12p70 production in α-GalCer-loaded but not upregulated after a 1 week cytokine treatment. En- MAGE-A4 peptide-loaded DCs. The importance of graftment of HLA-DRB1*08:03-positive primary the α-GalCer recognition by the intrinsic invariant leukemic stem cells in non-obese diabetic/severe γ TCR implies that the additional transfer of the combined immunodeficient/ c-null (NOG) mice α-GalCer-loaded DCs is beneficial for induction of was completely inhibited by in vitro preincubation IL-12p70 in vivo, leading to an improved efficacy of cells with CTL-1H8, suggesting that of cellular immunity. HLA-DRB1*08:03 is expressed on leukemic stem cells. Finally, analysis of the precursor frequency of In summary, tumor-reactive TCR transfer of + iNKT cells with both cytotoxic activity and adju- CD8 CTL specific for recipient antigens in vant properties is a powerful strategy for applica- post-HSCT peripheral blood T cells revealed a sig- tion as adoptive immunotherapy of cancer. nificant fraction of total donor CTL responses towards individual mismatched HLA-DR antigens in

31 two patients. These findings underscore unexpect- their genetic data. The P2RX7 gene on chromosome edly significant CD8 T cell responses in the context 12q was found to encode the minor H antigen rec- of HLA class II. It remains to be determined in fu- ognized by a CTL clone inducing severe pulmonary ture studies whether targeting a mismatched GVHD. Analysis of P2RX7 minigenes and syn- HLA-DR molecule, especially late after HSCT thetic P2RX7-derived peptides defined the when inflammatory conditions have subsided, nonameric P2RX7265-273 peptide WFHHCHPKY. might induce detrimental GVHD. In addition, po- Differential recognition of antigen-positive and tential targeting of an HLA-DP molecule whose -negative cells proved attributable to the nonsyn- disparity is almost permissive following HSCT onymous A↔G SNP rs7958311, which creates a should be examined. His↔Arg amino acid polymorphism at residue 270 in P2RX7. Real-time PCR showed P2RX7 to be *1 Department of Obstetrics and Gynecology, Nagoya highly expressed in spleen, ovary, lung, and pe- University Graduate School of Medicine. *2 ripheral blood cells, with particularly high levels in Department of Immunology, The University of Texas CD14+ blood cells. Immunohistochemistry of lung MD Anderson Cancer Center. tissue revealed P2RX7 protein expression restricted *3 Department of Hematology, Japanese Red Cross Na- goya First Hospital. to the alveolar epithelium, suggesting that unex- *4 Department of Pediatrics and Neonatology, Nagoya pected expression of P2RX7 transcripts in the lung City University, Graduate School of Medical Science. was attributable to pulmonary GVHD. The results *5 Central Institute for Experimental Animals. of these studies illustrate the potential to selectively *6 Department of Hematology and Cell Therapy, Aichi enhance graft-versus-leukemia activity by adoptive Cancer Center Hospital. transfer of minor H antigen-specific T-cell clones *7 Aichi Comprehensive Health Science Center, Aichi after careful selection of target antigens and CTL Health Promotion Foundation. dose escalation. *8 Division of Hematology, Fujita Health University, School of Medicine. *1 Division of Hematology, Fujita Health University, School of Medicine. 5. Identification of minor histocompatibility *2 Program in Immunology, Clinical Research Division, antigens responsible for pulmonary Fred Hutchinson Cancer Research Center *3 graft-versus-host disease following Department of Hematology/Oncology, Graduate School of Medicine, University of Tokyo. adoptive immunotherapy *4 *1 *2 *2 21st Century COE Program, Graduate School of Medi- Akatsuka, Y. , Warren, E.H. , Fujii, N. , Brown, *2 *3, 4, 5 *3 cine, University of Tokyo. M.L. , Ogawa, S. , Matsubara, A. , and Riddell, *5 *2 Core Research for Evolutional Science and Technol- S.R. ogy, Japan Science and Technology Agency. The adoptive transfer of donor T cells that rec- ognize recipient minor histocompatibility (H) anti- 6. Role of type 1 IFNs in antiglioma gens has potential as a strategy for preventing or immunosurveillance - using mouse treating leukemic relapse after allogeneic hema- studies to guide examination of novel topoietic cell transplantation (HSCT). Patients with prognostic markers in humans recurrent leukemia after HLA-matched allogeneic *1 *2 Fujita, M., Scheurer, M.E. , Decker, S.A. , McDonald, HSCT were treated with infusions of donor-derived, H.A.*3, Kohanbash, G.*3, Kastenhuber, E.R.*3, Kato, H.*3, ex vivo-expanded CD8+ cytotoxic T lymphocyte Bondy, M.L.*4, Ohlfest, J.R.*2, and Okada, H.*3 (CTL) clones specific for tissue-restricted recipient We have hypothesized that type-1 interferons minor H antigens at Fred Hutchinson Cancer Re- (IFN) play a pivotal role in anti-glioma immuno- search Center. Molecular characterization of the surveillance through promotion of type-1 adaptive minor H antigens recognized by CTL clones ad- immunity and suppression of immunoregulatory ministered to 3 patients was performed to provide cells. To evaluate this hypothesis, we induced de insight into the anti-leukemic activity and safety of -/- novo gliomas in Ifnar1 (deficient for type-1 IFN T-cell therapy. Using CTL clones that might be as- receptors) or wild type (WT) mice by intracerebral sociated with pulmonary toxicity following CTL transfection of NRAS and a short hairpin RNA infusion in 3 patients, which was severe in 1 patient against P53 using the Sleeping Beauty (SB) trans- at a higher CTL dose level, we first identified genes -/- poson system. Ifnar1 mice exhibited accelerated encoding the respective minor H antigens and ex- tumor growth and death. Analyses of their brain amined their levels of expression in lung tissue. tumor-infiltrating lymphocytes revealed an increase Associations were assessed by means of Interna- + + of cells positive for CD11b Ly6G and tional HapMap Project B-lymphoid cell lines and + + CD4 FoxP3 , markers which represent mye-

32 Fig. 1. Overall survival was evaluated in 304 glioma patients with grade 2-3 gliomas with reference to Fig. 2. Gliomas were induced in neonatal C57BL/6 the IFNAR1 rs1041868 genotype. Patients with mice by intraventricular transfection of the following AA genotype (red line) exhibited a significantly plasmids: pT2/C-Luc//PGK-SB1.3 (0.2 µg), shorter survival than those with the AG/GG geno- pT/CAG-NRas (0.4 µg), and pT/shp53 (0.4 µg). The types (black line). The P-value is based on the mice with growing tumors received daily treatment with ASA (solid line) or vehicle control (dashed line) log-rank test. as indicated. Symptom-free survival was moni- tored. The P-value is based on the log-rank test. myeloid-derived suppressor cells and regulatory “Med” in the tables stands for median survival. T-cells, respectively, but a decrease of CD8+ CTLs -/- as compared with WT mice. Ifnar1 mouse-derived 7. COX-2 blockade suppresses glioma tissues exhibited a decrease in mRNA for a gliomagenesis by inhibiting CTL-attracting chemokine CXCL10, but increases accumulation of myeloid-derived of CCL2 and CCL22, both of which are known to suppressor cells attract immunoregulatory cell populations. Den- Fujita, M., Kohanbash, G.*1, Fellows-Mayle, W.*1, dritic cells generated from bone marrow of Ifnar1-/- Hamilton, R.L.*1, Komohara, Y.*2, Decker, S.A.*3, mice failed to function as effective antigen present- Ohlfest, J.R.*3, and Okada, H.*1 ing cells. Moreover, depletion of Ly6G+ cells pro- Epidemiologic studies have highlighted associa- longed the survival of mice bearing gliomas. Fur- tions between regular use of nonsteroidal thermore, analyses of type-1 IFN-related sin- anti-inflammatory drugs (NSAID) and reduced gle-nucleotide polymorphisms (SNPs) in 291 pa- glioma risk in humans. Most NSAIDs function as tients with low-grade gliomas or anaplastic gliomas COX-2 inhibitors that prevent production of prosta- revealed that examples in IFNAR1 and IFNA8 are glandin E (PGE ). Because PGE induces expansion associated with significantly altered overall survival. 2 2 of myeloid-derived suppressor cells (MDSC), we Our new SB-induced murine glioma model led us to hypothesized that COX-2 blockade would suppress discover a pivotal role for the type-1 IFN pathway gliomagenesis by inhibiting MDSC development in anti-glioma immunosurveillance and relevant and accumulation in the tumor microenvironment human SNPs that may represent novel prognostic (TME). In mouse models of glioma, treatment with markers. the COX-2 inhibitors acetylsalicylic acid (ASA) or *1 Department of Pediatrics, Baylor College of Medicine, celecoxib inhibited systemic PGE2 production and USA delayed glioma development. ASA treatment also *2 Department of Pediatrics, University of Minnesota reduced the MDSC-attracting chemokine CCL2 Cancer Center, USA (C-C motif ligand 2) in the TME along with num- *3 Brain Tumor Program, University of Pittsburgh Can- bers of CD11b+Ly6GhiLy6Clo granulocytic MDSCs cer Institute, USA in both the bone marrow and the TME. In support *4 Department of Epidemiology, University of Texas MD of the conclusion that COX-2 blockade inhibited Anderson Cancer Center, USA systemic development of MDSCs and their CCL2-mediated accumulation in the TME, defects

33 in these processes were observed in glioma-bearing CXCL10 limited this pathway of immunosuppres- Cox2-deficient and Ccl2-deficient mice. Conversely, sion. Taken together, our findings show that the these mice and ASA-treated wild-type mice dis- COX-2 pathway promotes gliomagenesis by direct- played enhanced expression of CXCL10 (C-X-C ly supporting systemic development of MDSCs and motif chemokine 10) and infiltration of cytotoxic T their accumulation in the TME, where they limit lymphocytes (CTL) in the TME, consistent with a CTL infiltration. relief of MDSC-mediated immunosuppression. An- *1 tibody-mediated depletion of MDSCs delayed glio- Brain Tumor Program, University of Pittsburgh Can- cer Institute, USA ma growth in association with an increase in *2 CXCL10 and CTLs in the TME, underscoring a Department of Pathology, Kumamoto University critical role for MDSCs in glioma development. School of Medicine, JP *3 Department of Pediatrics, University of Minnesota Finally, Cxcl10-deficient mice exhibited reduced Cancer Center, USA CTL infiltration of tumors, providing evidence that

34 From left to right First row: Ms. Toshiko Gamano, Ms. Chieko Noda, Ms. Atsuko Sugimoto. Second row: Mr. Yohei Narita, Dr. Takayuki Murata, Dr. Tatsuya Tsurumi. Third row: Dr. Hiroki Isomura, Dr. Daisuke Kawashima, Dr. Sho Nakasu, Dr. Shinichi Saito, Dr. Teru Kanda.

35 Division of Virology ______

Tatsuya Tsurumi, M.D. Chief Teru Kanda, M.D. Section Head Hiroki Isomura, M.D. Section Head (until December 2011) Sho Nakasu, PhD. Senior Researcher Takayuki Murata, Vet. M.D. Researcher Sanae Nakayama, Ph.D. Research Resident (until March 2010) Shigeki Chiba, Ph.D. Research Resident (until March 2010) Shinichi Saito, Ph.D. Research Resident (as of April 2010) Daisuke Kawashima, Ph.D. Research Resident (as of May 2010) Chikako Sakuragi, Research Assistant (until March 2010) Chieko Noda, Research Assistant (as of April 2010) Naoe Hotta, Semi-regular employee (until May 2010) Toshiko Gamano, Semi-regular employee (as of June 2010)

Visiting Trainees Atsuko Sugimoto, Nagoya University Graduate School of Medicine Shigenori Toyama, Graduate School of Pharmaceutical Sciences, Nagoya City University Chieko Noda, Graduate School of Pharmaceutical Sciences, Nagoya City University Yohei Narita, Nagoya University Graduate School of Medicine

General Summary Approximately 15% of all human cancers have a viral etiology, but only seven viruses have actually been unequivocally implicated in neoplastic development. Among these the Epstein-Barr virus (EBV) is the primary object of our own studies. EBV is a ubiquitous gamma herpesvirus associated with several malignant diseases, including Burkitt’s lymphoma, nasopharyngeal lymphoma, a subset of Hodgkin’s lymphomas, some gastric cancers, and B cell lymphomas in immunosuppressed patients. Our research aims are to elucidate the molecular mechanisms of viral proliferation and oncogenesis of EBV as part of the world-wide effort to combat virus-infected cancers. During the period 2010-2011, our research interest was concentrated on the following issues: 1) Identification and characterization of CCAAT enhancer-binding protein (C/EBP) as a transcriptional activator for Epstein-Barr virus oncogene latent membrane protein 1; 2) Involvement of Jun dimerization protein 2 (JDP2) in the maintenance of Epstein-Barr virus latency; 3) Transcriptional repression by sumoylation of Epstein-Barr virus BZLF1 protein and its association with histone deacetylase; 4) Necessity of tetrameric ring formation of Epstein-Barr virus polymerase processivity factor for viral replication; 5) Participation of spatiotemporally differing DNA repair systems in Epstein-Barr virus genome maturation; 6) Unexpected instability of family of repeats (FR), the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems; 7) Assembly of human cytomegalovirus gene products essential for late viral gene expression into prereplication complexes before viral DNA replication.

1. Identification and characterization of lular signals, such as NF-B, MAPK, JAK/STAT CCAAT enhancer-binding protein and AKT. While transcription of LMP1 in latent (C/EBP) as a transcriptional activator type III cells is generally under the control of the for Epstein-Barr virus oncogene latent viral coactivator EBNA2, little is known about membrane protein 1 EBNA2-independent LMP1 expression in type II Noda, C., Murata, T., Kanda, T. and Tsurumi, T. latency. We thus screened a cDNA library for fac- Epstein-Barr virus LMP1, a major oncoprotein tors that can activate the LMP1 promoter in an expressed in latent infection, is critical for primary EBNA2-independent manner, using a reporter assay B cell transformation, functioning as a TNFR fa- system. So far, we have screened >20,000 clones, mily member by aggregation in the plasma mem- and newly identified C/EBP as a transcriptional brane and resulting in constitutive activation of cel- activator. Exogenous expression of C/EBP,

36 Figure 1. Proposed model for EBV LMP1 promoter activation by C/EBP. C/EBP binds to the functional binding site in the ED-L1 promoter region, and mediates activation of both TR-L1 and ED-L1 promoters. or efficiently augmented LMP1 mRNA and the ZII cis-element, a binding site for protein levels in EBV-positive cell lines, while CREB/ATF/AP-1. Suppression of Zp activity by other members of the C/EBP family exhibited only JDP2 correlated with HDAC3 association and re- modest or little activity. It has been demonstrated duced levels of histone acetylation. Although in- that LMP1 gene transcription depends on two pro- troduction of point mutations into the ZII element moter regions: proximal (ED-L1) and distal of the viral genome did not increase the level of (TR-L1). Interestingly, although we first used the BZLF1 production, silencing of endogenous JDP2 proximal promoter for screening, we found that gene expression by RNA interference increased the C/EBP increased transcription from both promoters levels of viral early gene products and viral DNA in latent EBV-positive cells. Mutagenesis in re- replication. These results indicate that JDP2 plays a porter assays and EMSA identified only one func- role as a repressor of Zp and that its replacement by tional C/EBP binding site, through which activation CREB/ATF/AP-1 at ZII is crucial to triggering re- of both proximal and distal promoters is mediated. activation from latency to lytic replication. Introduction of point mutations into the identified C/EBP site in EBV-BAC caused reduced LMP1 3. Transcriptional repression by transcription from both LMP1 promoters in epithe- sumoylation of Epstein-Barr virus lial cells. In conclusion, C/EBP is a newly identi- BZLF1 protein correlates with histone fied transcriptional activator of the LMP1 gene, in- deacetylase association dependent of the EBNA2 coactivator. Murata, T. and Tsurumi, T. The transition from latent to lytic phases of the 2. Involvement of Jun dimerization protein 2 Epstein-Barr virus life cycle is triggered by expres- (JDP2) in the maintenance of sion of a viral transactivator, BZLF1, that then in- Epstein-Barr virus latency duces expression of the viral immediate-early and Murata, T. and Tsurumi, T. early genes. The BZLF1 protein is Reactivation of the Epstein-Barr virus from post-translationally modified by a small ubiq- latency is dependent on expression of the BZLF1 uitin-related modifier-1 (SUMO-1). Here we found viral immediate-early protein. The BZLF1 promoter that BZLF1 is conjugated at lysine 12 not only by (Zp) normally exhibits only low basal activity but is SUMO-1 but also by SUMO-2 and 3. The K12R activated in response to chemical inducers such as mutant of BZLF1, which no longer becomes su- 12-O-tetradecanoylphorbol-13-acetate and calcium moylated, exhibited stronger transactivation than ionophore. We found that Jun dimerization protein the wild-type BZLF1 in a reporter assay system as 2 (JDP2) plays a significant role in suppressing Zp well as in the context of ther virus genome with nu- activity. Reporter, EMSA and ChIP assays of a Zp cleosomal structures. Furthermore, exogenous sup- mutant virus revealed JDP2 association with Zp at ply of a SUMO-specific protease, SENP, caused

37 de-sumoylation of BZLF1 and enhanced creased as lytic replication progressed. Based on BZLF1-mediated transactivation. Immunoprecipita- these observations, we conclude that sumoylation of tion experiments proved histone deacetylase 3 to be BZLF1 regulates its transcriptional activity through preferentially associated with the sumoylated form histone modification during Epstein-Barr virus of BZLF1. Levels of the sumoylated BZLF1 in- productive replication.

4. Tetrameric ring formation of Epstein-Barr virus polymerase processivity factor is crucial for viral replication Nakayama, S. and Tsurumi, T. The Epstein-Barr virus BMRF1 DNA polymerase processivity factor, which is essential for viral genome replication, exists mainly as a C-shaped head-to-head homodimer but partly forms ring-shaped tetramers through tail-to-tail association. Based on its molecular structure, several BMRF1 mutant viruses were constructed to exam- ine their influence on viral replication. The R256E virus, which has a severely impaired capacity for DNA binding and polymerase processivity, failed to form replication compartments, resulting in interference with viral replication, while the C95E mutation, which impairs head-to-head contact in vitro, unexpectedly hardly affected the viral replication. Also surprisingly, replication of the C206E virus, which would be expected to impair tail-to-tail contact, was severely restricted, although the mutant protein possesses the same in vitro biochemical activity as the wild type. Since the tail-to-tail contact surface is smaller than that of the head-to-head contact area, its contribution to ring formation might be essential for viral replication.

5. Spatiotemporally different DNA repair systems participate in Epstein-Barr virus genome maturation Sugimoto, A. and Tsurumi, T. Productive replication of Epstein-Barr virus occurs in discrete sites in nuclei, called replication compartments, where viral DNA replication proteins and host homologous recombinational repair (HRR) and mismatch repair (MMR) factors are recruited. Three-dimensional (3D) surface reconstruc- Figure 2. Mutated amino acid residues of EBV tion imaging clarified the spatial arrangements of BMRF1 (amino acids [aa] 1 to 314). (A) The these factors within the replication compartments. ring-shaped crystal structure of a tetramer of Subnuclear domains, designated BMRF1 cores C-terminally truncated BMRF1 protein (RCSB which are highly enriched in viral polymerase Protein Data Bank accession no. 2Z0L) is drawn processivity factor BMRF1, could be identified in- as a surface model, in which the mutated amino side the replication compartments. Pulse-chase ex- acid residues are displayed in color. (B) The mu- periments revealed that newly synthesized viral tated amino acid residues are displayed in color genomes organized around BMRF1 cores were on a gray surface model. The partner molecule forming a homodimer is drawn as a gray ribbon. transferred inward. HRR factors could be demon- The lower panel provides an alternative view of strated mainly outside BMRF1 cores, where de no- the complex. vo synthesis of viral DNA was ongoing, whereas MMR factors were found predominantly inside.

38 These results imply that de novo synthesis of viral DNA is coupled with HRR outside the cores, followed by MMR inside cores for quality control of 6. Unexpected instability of Family of replicated viral genomes. Thus, our approach un- Repeats (FR), the critical cis-acting veiled a viral genome manufacturing plant. sequence required for EBV latent infection, in EBV-BAC systems Kanda, T. and Tsurumi, T. A group of repetitive sequences, known as the Family of Repeats (FR), is a critical cis-acting sequence required for EBV latent infection. The FR sequences are heterogeneous among EBV strains, and they are sometimes subject to partial deletion when subcloned in E. coli-based cloning vectors. However, FR stability in the EBV-BAC (bacterial artificial chromosome) system has never been investigated. We found that full length FR of the Akata strain EBV was not stably maintained in a BAC vector. In contrast, newly obtained BAC clones of the B95-8 strain of EBV stably maintained the full length FR during recombinant virus production and B-cell transformation. Investigation of primary DNA sequences of Akata-derived EBV-BAC clones indicated that FR instability is most likely due to a putative secondary structure of the FR region. We conclude that the FR instability in EBV-BAC clones can be a pitfall in E. coli-mediated EBV genetics.

Figure 3. Newly synthesized viral DNA genomes localized outside the BMRF1 core move to the inside. Pulse-chase labeling experiments were performed with Tet-BZLF1/B95-8 cells at 24 h after induction. Outlines of the experimental protocol are given at the tops of panels A and B. (A) Tet-BZLF1/B95-8 cells were pulse-labeled with CldU for 10 min at 24 h postinduction. (B) The pulse-labeled cells were washed, and then chased for 1 h (B). For the 2D images, cells were treated with 0.5% mCSK buffer and stained with anti-BMRF1 or -BALF2 (green) and anti-CldU (red) antibodies. These images show bright- est-point projections of 60 images collected at 0.26-µm steps in the z axis. The same data are displayed as 3D topographical reconstructions of the BMRF1 or BALF2 protein and CldU-labeled viral DNA (left and middle panels, respectively). The right panels show 3D surface reconstruction images.

39 7. Human cytomegalovirus gene products were not detected unless the wild-type viral proteins essential for late viral gene expression were expressed in trans. At a high MOI, mutation of assemble into prereplication complexes ORF UL79, -87, or -95 had no effect on the level of before viral DNA replication major immediate-early (MIE) gene expression or Isomura, H. and Tsurumi, T. viral DNA replication, but late viral gene expres- Regulation of human cytomegalovirus (HCMV) sion from the UL44, -75, and -99 ORFs was not late gene expression by viral proteins is poorly detected. At a low MOI, preexpression of UL79 or understood, although such proteins could be targets -87, but not UL95, in human fibroblast cells nega- for novel antiviral agents. HCMV open reading tively affected the level of MIE viral gene expres- frames (ORFs) UL79, -87, and -95 encode proteins sion and viral DNA replication. The products of with homology to late gene transcription factors of ORFs UL79, -87, and -95 were expressed as early murine gammaherpesvirus 68 ORFs 18, 24, and 34, viral proteins and recruited to prereplication com- respectively. To determine whether these HCMV plexes (pre-RCs), along with UL44, before the ini- proteins are also essential for late gene transcription tiation of viral DNA replication. All three HCMV of a beta herpesvirus, we mutated HCMV ORFs ORFs appear indispensable for late viral gene ex- UL79, -87, and -95. Cells were infected with the pression and viral growth. The roles of UL79, -87, recombinant viruses at high and low multiplicities and -95 in pre-RCs for late viral gene expression of infection (MOIs). While viral DNA was detected clearly warrant further attention. with the recombinant viruses, infectious viruses

40 From left to right Dr. Yasushi Kojima, Dr. Masahiro Aoki, Dr. Keiichiro Sakuma, Ms. Yoshiko Goto

41 Division of Molecular Pathology ______

Reiji Kannagi, M.D., D.M.Sc., Chief (until March, 2010) Masahiro Aoki, M.D., D.M.Sc., Chief (as of December, 2010) Mamoru Kyogashima, M.D., D.M.Sc., Section Head (until March, 2011) Keiichiro Sakuma, M.D., D.M.Sc., Senior Researcher Akiko Kanamori, Ph.D., Senior Researcher (until March, 2010) Yasushi Kojima, M.D., D.M.Sc., Senior Researcher (as of April, 2011) Akiko Yusa, Ph.D., Research Resident (until March, 2011) Yoshiko Goto, D.V.M.S., Research Assistant Keiko Tamiya-Koizumi, Ph.D., Semi-regular Employee (until March, 2011)

Visiting Scientists Osamu Taguchi, D.M.Sc., (until March, 2011)

Visiting Trainees Kouji Tanaka, B.Ph., Nagoya City University (until March, 2011) Sachiko Kondo, M.E., Nagoya City University

General Summary The incidence of colon cancer is increasing steadily in Japan; the disease is predicted to become the most common cancer by 2020. Accumulating evidence suggests that in addition to genetic and epigenetic changes in the genome of cancer cells, interactions of cancer cells with non-cancer stromal cells play essential roles in support of colon cancer progression. Until November 2010, we were focused on the roles of cell adhesion molecules and cell surface glycan molecules, including sialyl Lewis X and sialyl Lewis A, in colon cancer. Our research since December 2010 has aimed mainly to identify novel molecular targets for prevention and/or therapy of colon cancer through careful analyses of the intestinal tumor progression in genetically engineered mouse models, followed by evaluation of the findings using clinical samples. We are currently interested in the following subjects: (1) Genetic dissection of the signaling pathways that play pivotal roles in colon carcinogenesis; (2) Elucidation of complex tumor-stroma interactions that promote invasion and metastasis of colon cancer; and (3) Iden- tification of novel metastasis suppressor genes in colon cancer.

1. Induction of selectin-ligand glycans ability to maintain colon cancer stem cells, during the epithelial-mesenchymal considered to have high metastatic ability. transition of colon cancer From these observations, we hypothesized x Sakuma, K., Aoki, M. and Kannagi, R. that EGF and bFGF might enhance sLe and Sialyl Lewis X (sLex) and sialyl Lewis A sLea expression. We were able to demon- (sLea) promote extravasation of cancer cells strate that EGF/bFGF-treated CRC cells in- via binding to E-selectin. High expression deed had markedly increased expression of levels of these glycans are correlated with a sLex and sLea, resulting in enhanced poor prognosis in cases of colorectal cancer E-selectin binding. RT-PCR analysis indi- (CRC). However, the regulatory mechanisms cated altered levels of glycogene?? expres- underlying their expression remain unclear. sion in the EGF/bFGF treated cells, which Clinically, levels of EGFR and FGFR are re- explained the elevated glycan expression. lated with the incidence of metastasis. Inter- Prolonged stability of c-Myc and estingly, EGF and bFGF are known for their down-regulation of CDX2 were associated

42 Fig. 1. Results of flow cytometric analysis indicating induction of selectin-ligand glycan expression on colon cancer cells by EGF/bFGF-treatment.

with these alterations. Furthermore, cancer cells. To elucidate molecular mecha- the treated cells exhibited drastic EMT-like nisms contributing to the impaired sulfation, changes. These results suggest that EGF and we studied the expression levels of mRNAs bFGF may confer metastatic ability to CRC for 6-O-sulfotransferase isoenzymes, PAPS cells by inducing EMT and expression of se- synthases and transporters, and the cell mem- lectin-ligand glycans in vitro. Since immuno- brane sulfate transporter DTDST in cancer histochemical study of clinical samples de- tissues. The most striking decrease in cancer tected CRC cells undergoing EMT and ex- cells compared with non-malignant epithelial a hibiting high sLe expression, our findings cells was noted for the mRNA level of may help to explain the molecular mecha- DTDST encoding a cell membrane sulfate nisms responsible for metastasis of CRC transporter (P = 0.0000014; n = 20). Most cells. cultured colon cancer cells showed reduced mRNA levels of DTDST, which were re- 2. Induction of sialyl Lewis x by stored on culture with histone deacetylase in- epigenetic silencing of DTDST hibitors such as butyrate. In contrast, inhibi- expression tors of DNA methylation, like Yusa, A., Miyazaki, K., Lim, K.T., Yin, J., Yagi, 5-aza-2-deoxycytidine, failed to induce H. and Kannagi, R. expression of DTDST mRNA. ChIP assays Sialyl Lewis x (sLex) is expressed on showed that silencing of the DTDST gene in cancer cells and promotes hematogenous cultured colon cancer cells was correlated metastasis via binding to E-selectin. Normal with histone H3 lysine 9 deacetylation and colon epithelial cells virtually do not express sLex, while they feature high levels of sialyl 6-sulfo Lex, a sulfated derivative of sLex. In contrast, colon cancer cells express sLex highly, while they exhibit markedly reduced expression of sialyl 6-sulfo Lex, compared with normal colon epithelial cells. These observations suggest impaired sulfation during the course of malignant transformation of colon epithelial cells, resulting in increased sLex expression in

Fig. 2. Induction of DTDST reduces the level of sLex and increases the level of sialyl 6-sulfo Lex. A, Results of RT-PCR indicating tet-off inducible DTDST transcription. B, DOX-treated colon cancer cells exhibit markedly decreased sLex and increased sialyl 6-sulfo Lex expression.

43 Fig.3. JNK activation stimu- lates mTOR signaling in the ∆716 intestinal polyps of Apc∆ mice. JNK can activate mTORC1 signaling through phosphorylation of Raptor. Activation of JNK helps increase the levels of both mRNA and protein associated with tumor growth through stimulation of c-Jun and mTORC1 signaling, respectively. loss of H3 lysine 4 methylation of the In the present study, we found that c-Jun DTDST gene promoter. Methylation of H3 N-terminal kinase (JNK) is involved in the lysine 9, H3 lysine 27, and H4 lysine 20 was mTORC1 activation. Both JNK and also noted in some cultured colon cancer mTORC1 showed tumor-size dependent cells with diminished DTDST transcription. activation in intestinal polyps of Apc 716 Finally, we demonstrated that induction of mice, and the JNK inhibitor SP600125 sig- DTDST expression resulted in reduction in nificantly suppressed the tumor formation. sLex and increase in sialyl 6-sulfo Lex. Un- The JNK activator anisomycin caused expectedly, the growth rate of the cancer mTORC1 activation in colon cancer cell lines, cells was markedly enhanced when DTDST whereas SP600125 and siRNAs against JNKs was suppressed. These results show that de- suppressed the signaling. JNK stimulated the crease in the transcription of the sulfate mTORC1 kinase activity in vitro through di- transporter gene is the major cause of de- x rect phosphorylation of Raptor at serine 863. creased expression of sialyl 6-sulfo Le and Interestingly, the mTORC1 inhibitor increased expression of sLex in colon cancer. RAD001 reduced the protein levels of cyclin The results also suggest that diminished E, proliferin, and osteopontin, while knock- DTDST expression is closely related to en- down of c-Jun or by SP600125 reduced their hanced proliferation of cancer cells. mRNA levels. These results suggest that JNK may promote intestinal tumor formation by 3. Roles of the JNK/mTORC1 signaling inducing expression of growth regulatory in intestinal tumorigenesis in Apc proteins via two mechanisms: (1) enhanced mutant mice *1 *1 transcription via phosphorylation of c-Jun; Aoki, M., Fujishita, T. and Taketo, MM. and (2) increased mRNA translation via Most human colorectal cancers exhibit activation of mTORC1. somatic mutations in the adenomatous polyposis coli (APC) tumor suppressor gene, *1 Department of Pharmacology, Graduate School leading to Wnt signaling activation, and loss of Medicine, Kyoto University of APC function is considered the earliest event in multi-step colon cancer progression. Apc716 mice develop numerous adenomatous polyps in the intestines, and serve as a valuable model that can effectively recapitu- late human colon tumorigenesis. We previously reported activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling in intestinal tumors of Apc716 mice.

44 From left to right First row: Ms. E. Kawamoto, Dr. M. Inagaki, Ms. Y. Takada Second row: Dr. Y. Hayashi, Ms. Z. Wang, Ms. C. Yuhara, Ms. K. Kobori Third row: Dr. K. Kasahara, Dr. P. Li, Dr. H. Goto, Dr. A. Inoko Forth row: Dr. I. Izawa, Dr. M. Matsuyama, Dr. D. He, Dr. H. Tanaka

45 Division of Biochemistry ______

Masaki Inagaki, M.D., Ph.D. Chief Ichiro Izawa, M.D., Ph.D. Section Head Hidemasa Goto, M.D., Ph.D. Section Head Akihito Inoko, M.D., Ph.D. Senior Researcher Kousuke Kasahara, Ph.D. Researcher Yuko Hayashi, Ph.D. Research Assistant Takashi Shiromizu, Ph.D. Research Resident (until March 2010) Miho Ibi, D.D.S., Ph.D. Research Resident (until March 2010) Ping Li, M.D., Ph.D. Research Resident (until March 2011) Makoto Matsuyama, Ph.D. Research Resident (as of April 2011) Dongwei He, Ph.D. Research Resident (as of April 2011) Hiroki Tanaka, D.D.S., Ph.D. Research Resident (as of May 2011) Kyoko Kobori, Semi-regular Employee Eriko Kawamoto, Semi-regular Employee (as of April 2011)

Visiting Scientists Miho Ibi, D.D.S., Ph.D. Global COE Postdoctoral Fellow, Nagoya University Graduate School of Medicine (as of April 2010 until March 2011) Ping Li, M.D., Ph.D. Global COE Postdoctoral Fellow, Nagoya University Graduate School of Medicine (as of April 2011)

Visiting Trainees Yuki Ohmuro-Matsuyama, Ph.D. Research Resident of the Foundation for Promotion of Cancer Research Makoto Matsuyama, Ph.D. Research Resident of the Foundation for Promotion of Cancer Research Masato Enomoto, Ph.D. Department of Cellular Oncology, Graduate School of Medicine, Nagoya University

General Summary How cells coordinate proliferation and differentiation is a fundamental problem in cell and developmental biology, as well as being prominent altered features in cancer. As cells progress toward cell division, a number of cell-cycle checkpoints ensure that cells do not prematurely undergo cell-cycle transitions such as exit from G0 and entry into S phase or mitosis. In the normal development of multi-cellular organisms, signals controlling these cell-cycle checkpoints are thought to come from both intrinsic (e.g. centrosomes or cell polarity signaling) and extrinsic (e.g. cell-cell contacts or planar cell polarity signaling) sources. We are seek to define the mechanisms by which cells link the polarity signaling (tissue architecture) to cell-cycle control. Identification of such regulatory mechanisms and their failure to operate in cancer cells should contribute to novel strategies for cancer therapy. Our attention is focused on 2 specific areas: (1) Identification and functional analysis of protein kinases involved in cell cycle checkpoints; (2) Regulation of cytoskeletal proteins (especially intermediate filaments) and associated elements active in cell adhesion and determination of cell polarity. Specific projects/results are described in detail below.

1. Novel regulation of Checkpoint kinase 1 to activate DNA damage/replication checkpoints. In (Chk1) fact, Chk1 inhibition induces mitotic cell death Goto, H., Kasahara, K., Enomoto, M., Matsuyama, M., (mitotic catastrophe) under conditions of DNA Li, P., Tomono, Y*1., Kiyono, T*2. and Inagaki, M. damage or replication stress. Since Chk1 inhibitors Defects in p53 and/or pRb (retinoblastoma pro- have been demonstrated to sensitize tumors to a tein) function occur frequently in human cancers variety of DNA-damaging agents or due to mutations in or deletion of TP53 and/or RB1 anti-metabolites in preclinical models a number of or down-regulation of their associated pathways. examples are now being evaluated in Phase I or II Such defects render cancer cells dependent on Chk1 clinical trials.

46 In spite of the importance of Chk1 for molecular target therapy, the means by which it is regulated had remained largely unknown. However, Chk1 phosphorylation at Ser317 and Ser345 occurs with ATR (ataxia telangiecta- sia mutated- and rad3-related) and over the last 5 years we have therefore focused on Chk1 regulation by other protein kinases and elucidated related mechanisms. First, we showed significance of phosphorylates Chk1 specifically at Ser280. Chk1-Ser296 autophosphorylation after DNA Although Chk1 constantly shuttles between the cy- damage. After ATR-induced phosphorylation of toplasm and nucleus, Ser280 phosphorylation pro- Chk1 (by implication, catalytic activation of Chk1) motes nuclear retention of Chk1. Such nuclear ac- dephosphorylation is then promoted at the ATR cumulation contributes greatly to abrogation of the sites. A phosphorylation shift from ATR sites to DNA damage checkpoint. Since the Ras-MAPK Ser296 not only plays an important role in the pathway is up-regulated in a wide spectrum of hu- spread of Chk1 signals throughout the nucleoplasm man cancers, our observations point to a possibility but also induces Chk1 binding to 14-3-3, which that the p90 RSK-Chk1 pathway may serve as a serves as a platform between Cdc25A and barrier to protect genomic integrity in the case of Ser296-phosphorylated Chk1. Such ternary com- Ras-MAPK up-regulation. plex formation enables phosphorylation of Cdc25A Our studies have shown that Chk1 regulation is at Ser76 by Chk1, which causes generation of a more complex than previously considered. Further βTrCP recognition motif (phosphodegron) by analyses are required to evaluate Chk1 as a molecular target for cancer therapy. NEK11. This in turn results in protea- some-dependent Cdc25A degradation, preventing *1 Division of Molecular and Cell Biology, Shigei premature activation of cyclin-dependent kinases Medical Research Institute, Japan (Cdks) and inducing cell cycle arrest after DNA *2 Division of Virology, National Cancer Center Re- damage. search Institute, Japan Second, we demonstrated the importance of a novel positive feedback loop between Cdk1 and 2. Novel mitotic signaling from Akt/PKB to Chk1 at the G2/M transition. Before mitosis (at S or Polo-like kinase 1 G2 phase), Chk1 has basal activity in unperturbed Kasahara, K., Goto, H., Izawa, I., Kawamoto E., cells. Since Chk1 is predominantly localized in the Watanabe, N.*1, Tomono, Y.*2, Kiyono, T.*3 and Inagaki, nucleus of interphase cells, it inhibits nuclear Cdc25 M. phosphatase activity and thus blocks premature Chromosomal instability is a hallmark of ag- Cdk1 activation in interphase cells. Once initial ac- gressive malignancies. Recent analyses have tivation of Cdk1 occurs (likely in late G2), Cdk1 provided evidence that chromosomal instability it- starts to phosphorylate Chk1 at Ser286 and Ser301. self in fact plays important roles in tumour initia- This phosphorylation induces Chk1 translocation tion and/or progression. In support of this concept, from the nucleus to the cytoplasm. The elimination accumulating data have demonstrated a direct rela- of Chk1 kinase activity from the nucleus triggers a tionship between chromosomal stability and several positive feedback loop of Cdk1 activation in the tumour suppressor gene products, such as p53, pRb nucleus, which promotes mitosis. (regulators at the G1/S transition), Adenomatous Third, we have elucidated the consequences of Polyposis Coli (APC) protein (a regulator in Wnt Chk1 phosphorylation at Ser280 by p90 RSK (90 signalling) and BRCA2 (a regulator in DNA repair kDa ribosomal S6 kinase) downstream of growth signalling). The PI3K-Akt signalling pathway trig- factor stimulation. Following stimulation of recep- gers a cascade of responses from cell growth and tor tyrosine kinase with growth factor, p90 RSK is proliferation, at the same time enhancing cell sur- activated downstream of the MAPK (mito- vival and motility, which drive tumour initiation gen-activated protein kinase) cascade and then

47 and progression. However, its relationship to chro- ously demonstrated site-specific phosphorylation to mosomal instability remains unknown. induce disassembly of vimentin filaments in vitro, Polo-like kinase 1 (Plk1) is an essential kinase and that in cultured cells, mitotic vimentin phos- regulating multiple aspects of mitosis. It has been phorylation is essential for efficient segregation of reported to be catalytically activated through phos- vimentin filaments into daughter cells. However, phorylation at Thr210 by Aurora A and that hBora, there is little information on the significance of this which accumulates toward the G2/M transition, is process in vivo. essential for this phosphorylation. In the present To clarify the biological function of vimentin study we found that Akt controls Plk1 functions in phosphorylation in vivo, we generated knock-in mitosis. Akt induces Plk1 phosphorylation at Ser99, mice with point mutations of mitotic phosphoryla- which creates a 14-3-3γ-binding site on Plk1. The tion sites in the head domain of vimentin. Here, we binding between Plk1 and 14-3-3γ is required for demonstrated that mice homozygous for mutations Plk1-induced phosphorylation of known substrates showed reduced lens size and increased cataract such as BubR1 and Wee1 in mitosis. Disturbance of formation. Histologic examination revealed a de- this process induces prometaphase or metaphase crease in the number of lens epithelial cells, associ- arrest, as does Plk1 depletion. Akt can phosphory- ated with disorganization of the lens fiber cells in late Ser99 on Plk1 only in the presence of hBora. both anterior and equatorial regions. Immunofluo- This accounts for the mitosis-specific phosphoryla- recence analyses critically demonstrated an emer- tion of Plk1-Ser99 by Akt. Therefore, Akt induces gence of multinucleated cells in lens epithelia, sug- Plk1 association with 14-3-3γ which then elevates gesting that the lens epithelial cells were not able to Plk1 activity. divide completely in mitosis. Moreover, we found In contrast to the abundant data regarding the polyploidy in the lens fiber cells of homozygous PI3K-Akt pathway, evidence for amplification or mutant mice. Taken together, these results indicate mutation of PLK1 in human tumours or tumour cell that mitotic vimentin phosphorylation is indis- lines is minimal. It is likely that PLK1 itself is not pensable for cells to divide efficiently, and thus is an oncogene. However, Plk1 is overactivated in a required for lens morphogenesis. variety of human cancers and this is mostly associ- Our study demonstrated a novel mechanism in ated with a poor prognosis. In addition, Plk1 is con- the requirement of mitotic vimentin phosphoryla- sidered to control spindle formation, probably tion for maintenance of lens morphogenesis. The through both centrosome maturation and formation observed relationship between cataract formation of stable kinetochore–microtubule attachments. and mitotic defects could open a new field to study Therefore, the Akt-Plk1 pathway is likely to play a control of lens morphology. critical role in promoting chromosomal instability in cancer cells in which the PI3K-Akt pathway is 4. Trichoplein blocks aberrant primary cilia up-regulated. assembly in proliferating cells Inoko, A., Matsuyama, M., Goto, H., Hayashi, Y., Ibi, *1 Chemical Library Validation Team, Chemical Biology M., He, D., Izawa, I., Urano, T.*1, Yonemura, S. *2, Department, RIKEN Advanced Science Institute (ASI) Kiyono, T. *3 and Inagaki, M. *2 Department of Molecular and Cell Biology, Shigei The primary cilium is an antenna-like organelle Medical Research Institute *3 which modulates differentiation, sensory functions Division of Virology, National Cancer Center Re- and signal transduction. Once cilia are disassembled search Institute at G0/G1 transition, ciliary reproduction is strictly inhibited in proliferating cells. However, the 3. Functional analyses of vimentin mechanisms responsible for this inhibition remain phosphorylation in mice largely unknown. Matsuyama, M., Tanaka Hir., Kobori K., Izawa I., and We previously reported that trichoplein, a novel Inagaki M. keratin-binding protein, localizes to the centriole in Intermediate filaments (IFs), together with actin proliferating cells, in addition to its localization on filaments and microtubules, form the cytoskeletal cell-cell contacts in polarized epithelial cells. Re- network maintaining cellular integrity. Vimentin is cently, we found that the centriolar trichoplein dis- the most abundant IF protein expressed in mesen- appears from ciliated mother centrioles in quiescent chymal cells and also in a variety of tumors. Cyto- cells. Exogenous expression of trichoplein inhibited plasmic vimentin is reorganized dramatically during primary cilia assembly in serum-starved cells mitosis, and this reorganization is considered to be whereas RNAi-mediated depletion accelerated as- controlled by protein phosphorylation. We previ- sembly under cultivation with serum. In vitro and in

48 vivo kinase assays revealed that trichoplein controls Aurora A (AurA) activation at the centrioles predominantly in G1 phase. In vitro analyses further confirmed that trichoplein binds and activates AurA directly. Using trichoplein fragments and mutants, we could demonstrate that 1-130 a.a. of trichoplein is a minimum requirement for centriolar localization and blocking cilia assembly. However, the mutant with Ala52 and Trp54 to Asp, featuring weak- ened binding to AurA, failed to block cilia assembly despite its centriolar localization. Collectively, our results indicate that suppression of primary cilia assembly by trichoplein necessitates not only the ability to localize to centrioles but also binding and activation of AurA. With respect to cell cycle regulation, trichoplein- or AurA-knockdown also induced G1 arrest but this phenotype was reversed by simultaneous knockdown of IFT20, a treatment resulting in an inability to form cilia in cells. These data suggest that the trichoplein-AurA pathway is integral to continuous suppression of primary cilia assembly, which is required for G1 progression.

*1 Department of Biochemistry, Shimane University School of Medicine *2 RIKEN Center for Developmental Biology *3 Virology Division, National Cancer Center Research Institute

49 From left to right Dr. E. Ito, Dr. H. Nakamura, Mr. Ha. Tanaka, Mr. Y. Nishikawa, Mr. Y. Minoura, and Dr. H. Kumimoto.

50 Central Service Unit ______

Masaki Inagaki, M.D. Chief Hiroshi Kumimoto, Ph.D. Senior Researcher Yasushi Minoura, B.P., Senior Research Assistant Harunari Tanaka, B.P. Research Assistant Yasuhiro Nishikawa, Research Assistant

Visiting Trainee Erika Ito, M.D. Nagoya City University, Graduate School of Medical Sciences (as of October 2010)

General Summary Our main research project concerns molecular epidemiologic analysis of human esophageal cancer. In particular, we are focusing on the relationship between numbers of polymorphisms in mitochondrial DNA (mtDNA) and risk of esophageal cancer development.

1.Mitochondria DNA polymorphism the cells. This elevated ROS may introduce muta- Kumimoto, H. tions into mtDNA and genomic DNA. We there- Mitochondria are the organelles responsible for fore have analyzed a number of polymorphisms in producing energy in cells but they also play major mtDNA as surrogate markers of ROS with a view roles in committing cells to apoptosis. Recently, to evaluating relationships with risk of esophageal mutations in mitochondrial DNA (mtDNA) have cancer. been found in various of cancers, including exam- We performed sequencing analysis of D-loop ples in the breast and stomach. Previously, we region in mtDNA using DNA samples from found frequent somatic mutations in the D-loop re- esophageal cancer subjects and non-cancer controls, gion in esophageal tumors (in 34 % of cases). We collected in the HERPACC study. With also assessed nuclear genomic instability, but did re-sequencing primers sets made by ABI, mito- not find any correlation with somatic mtDNA muta- SEQr the entire D-loop region could be se- tions, suggesting that instability of mtDNA in quenced. After sequencing, I identified polymor- esophageal cancer might be independent of nuclear phisms by comparison with the common mtDNA genomic instability. sequence. Energy as ATP is produced in mitochondria with The initial data for 24 esophageal cancer patients reactive oxygen species (ROS) as a byproduct. and 29 non-cancer controls, which demonstrated Polymorphisms in the genes related to oxidative totals of 12.3 and 12.0 on average, respectively phosphorylation may elevate ROS production by (Table 1). leaking electron. Therefore, the number of polymorphisms in mtDNA might influence ROS level in

Table 1. Summary of analyses of polymorphism in mtDNA Total Number of subjects Insertion Deletion SNP polymorphisms analyzed

Esophageal 12.3 1.8 0.67 9.8 24 cancer

Non-cancer 12.0 1.7 0.41 9.9 29

Data are expressed as average numbers on each subject, except for the 'Number of subjects analyzed'.

51 Librarians ______

From Left to right Ms. T. Shibata, Ms. T. Yasuda, Ms. M. Sakou, Ms. K. Horayama, supporting scientific and medical information.

52 Publications ______publication, 2011. (PMID: 22080964) Journals J008. Bargagna-Mohan, P., Paranthan, R.R., Hamza, A., Dimova, N., Trucchi, B., Srinivasan, J001. Akita, K., Yoshida, S., Ikehara, Y., C., Elliott, G.I., Zhan, C.G., Lau, D.L., Zhu, H., Shirakawa, S., Toda, M., Inoue, M., Kitawaki, J., Kasahara, K., Inagaki, M., Cambi, F., Mohan, R.: Nakanishi, H., Narimatsu, H., Nakada, H.: Withaferin A targets intermediate filaments glial Different levels of Sialyl-Tn Antigen expressed on fibrillary acidic protein and vimentin in a model of MUC16 in Endometriosis and Ovarian Cancer retinal gliosis. J Biol Chem, 285: 7657-7669, 2010. Patients. Int J Gynecol Cancer, in press. (PMID: 20048155) J002. Ali, A.H., Takizawa, H., Kondo, K., J009. Bhattacharyya, S., Deb, J., Patra, A.K., Matsuoka, H., Toba, H., Nakagawa, Y., Kenzaki, Thuy, Pham, D.A., Chen, W., Vaeth, M., K., Sakiyama, S., Kakiuchi, S., Sekido, Y., Sone, S., Berberich-Siebelt, F., Klein-Hessling, S., Lamperti, Tangoku, A.: 5-Aminolevulinic acid-induced E.D., Reifenberg, K., Jellusova, J., Schweizer, A., fluorescence diagnosis of pleural malignant tumor. Nitschke, L., Leich, E., Rosenwald, A., Brunner, Lung Cancer, 74: 48-54, 2011. (PMID: 21353719) C., Engelmann, S., Bommhardt, U., Avots, A., J003. An, B., Kondo, Y., Okamoto, Y., Shinjo, K., Muller, M.R., Kondo, E. Serfling, E.:NFATc1 Kanemitsu, Y., Komori, K., Hirai, T., Sawaki, A., affects mouse splenic B cell function by controlling Tajika, M., Nakamura, T., Yamao, K., Yatabe, Y., the calcineurin-NFAT signaling network. J Exp Fujii, M., Murakami, H., Osada, H., Tani, T., Med, 208: 823-839, 2011. (PMID: 21464221) Matsuo, K., Shen, L., Issa, J.P., Sekido, Y.:A J010. Boffetta, P., McLerran, D., Chen, Y., characteristic methylation profile in CpG island Inoue, M., Sinha, R., He, J., Gupta, P.C., Tsugane, methylator phenotype-negative distal colorectal S., Irie, F., Tamakoshi, A., Gao, Y.T., Shu, X.O., cancers. Int J Cancer, 127: 2095-2105, 2010. Wang, R., Tsuji, I., Kuriyama, S., Matsuo, K., (PMID: 20131317) Satoh, H., Chen, C.J., Yuan, J.M., Yoo, K.Y., J004. An, J., Fujiwara, H., Suemori, K., Niiya, Ahsan, H., Pan, W.H., Gu, D., Pednekar, M.S., T., Azuma, T., Tanimoto, K., Ochi, T., Akatsuka, Sasazuki, S., Sairenchi, T., Yang, G., Xiang, Y.B., Y., Mineno, J., Ozawa, H., Ishikawa, F., Nagai, M., Tanaka, Hid., Nishino, Y., You, S.L., Kuzushima, K., Yasukawa, M.: Activation of T-cell Koh, W.P., Park, S.K., Shen, C.Y., Thornquist, M., receptor signaling in peripheral T-cell lymphoma Kang, D., Rolland, B., Feng, Z., Zheng, W., Potter, cells plays an important role in the development of J.D.: Body mass index and diabetes in Asia: a lymphoma-associated hemophagocytosis. Int J cross-sectional pooled analysis of 900,000 Hematol, 93: 176-185, 2011. (PMID: 21229399) individuals in the Asia cohort consortium. PLoS One, 6: e19930, 2011. (PMID: 21731609) J005. Antonov, AS., Antonova, GN., Fujii, M., Dijke, PT., Handa, V., Catravas, JD., Verin, AD.: J011. Cai, Q., Long, J., Lu, W., Qu, S., Wen, W., Regulation of endothelial barrier function by TGF-β Kang, D., Lee, J.Y., Chen, K., Shen, H., Shen, type I receptor ALK5: Potential role of contractile C.Y., Sung, H., Matsuo, K., Haiman, C.A., Khoo, mechanisms and heat shock protein 90. J Cell U.S., Ren, Z., Iwasaki, M., Gu, K., Xiang, Y.B., Physiol, 227: 759-71, 2012. (PMID: 21465483) Choi, J.Y., Park, S.K., Zhang, L., Hu, Z., Wu, P.E., Noh, D.Y., Tajima, K., Henderson, B.E., Chan, J006. Aoki, K., Kakizaki, F., Sakashita, H., K.Y., Su, F., Kasuga, Y., Wang, W., Cheng, J.R., Manabe, T., Aoki, M., Taketo, M.M.: Suppression Yoo, K.Y., Lee, J.Y., Zheng, H., Liu, Y., Shieh, of colonic polyposis by homeoprotein CDX2 Y.L., Kim, S.W., Lee, J.W., Iwata, H., Le, through its nontranscriptional function that Marchand. L., Chan, S.Y., Xie, X., Tsugane, S., stabilizes p27Kip1. Cancer Res, 71: 593-602, 2011. Lee, M.H., Wang, S., Li, G., Levy, S., Huang, B., (PMID: 21224344) Shi, J., Delahanty, R., Zheng, Y., Li, C., Gao, Y.T., J007. Asai, D., Osone, S., Imamura, T., Shu, X.O., Zheng, W.: Genome-wide association Sakaguchi, H., Nishio, No., Kuroda, H., Kojima, study identifies breast cancer risk variant at S., Hosoi, H.: Allo-SCT in a patient with CRMCC 10q21.2: results from the Asia Breast Cancer eith aplastic anemia using a reduced intensity Consortium. Hum Mol Genet, 20: 4991-4999, 2011. conditioning regimen. Bone Marrow Transplant, in (PMID: 21908515)

53 J012. Cai, Q.Y., Wen, W.Q., Qu, S.M., Li, G.L., consortium. Front Cancer Epidemiol Prev, 1: 1-15, Egan, K.M., Chen, K., Deming, S.L., Shen, H., 2011. Shen, C.Y., Gammon, M., Blot, W.J., Matsuo, K., J017. Deguchi, A., Miyoshi, H., Kojima, Y., Haiman, C.A., Khoo, U.S., Iwasaki, M., Santella, Okawa, K., Aoki, M., Taketo, M.M.: LKB1 R., Zhang, L., Fair, A.M., Hu, Z.B., Wu, P.E., suppresses p21-activated kinase-1 (PAK1) by Signorello, L.B., Titus-Ernstoff, L., Tajima, K., phosphorylation of Thr109 in the p21-binding Henderson, B.E., Chan, Y.K., Kasuga, Y., domain. J Biol Chem, 285: 18282-18290, 2010. Newcomb, .P.A, Zheng, H., Cui, Y., Wang, F., (PMID: 20400510) Sheeh, Y.L., Iwata, H., Le Marchand, L., Chan, S.U., Shrubsole, M.J., Trentham-Dietz, A, J018. Ebisuno, Y., Katagiri, K., Katakai, T., Tsugane, S., Garcia-Closas, M., Long, J.R., Li, Ueda, Y., Nemoto, T., Inada, H., Nabekura, J., Chun, Shi, J.J., Huang, B., Xiang, Y.B., Gao, Y.T., Okada, T., Kannagi, R., Tanaka, T., Miyasaka, M., Lu, W., Shu, X.O., Zheng, W.: Replication and Hogg, N., Kinashi, T.: Rap1 controls lymphocyte functional genomic analyses of the breast cancer adhesion cascade and interstitial migration within susceptibility locus at 6q25.1 generalize its lymph nodes in RAPL-dependent and -independent importance in women of Chinese, Japanese and manners. Blood, 115: 804-814, 2010. (PMID: European ancestry. Cancer Res, 71: 1344-1355, 19965628) 2011. (PMID: 21303983) J019. Ennishi, D., Asai, H., Maeda, Y., J013. Cheng, A.S., Lau, S.S., Chen, Y., Kondo, Shinagawa, K., Ikeda, K., Yokoyama, M., Terui, Y., Y., Li, M.S., Feng, H., Ching, A.K., Cheung, K.F., Takeuchi, K., Yoshino, T., Matsuo, K., Hatake, K., Wong, H.K., Tong, J.H., Jin, H., Choy, K.W., Yu, Tanimoto, M.: Statin-independent prognosis of J., To, K.F., Wong, N., Huang, T.H., Sung, J.J.: patients with diffuse large B-cell lymphoma EZH2-mediated concordant repression of Wnt receiving rituximab plus CHOP therapy. Ann Oncol, antagonists promotes β-catenin-dependent 21: 1217-1221, 2010. (PMID: 19884246) hepatocarcinogenesis. Cancer Res, 71: 4028-4039, J020. Ennishi, D., Maeda, Y., Niitsu, N., Kojima, 2011. (PMID: 21512140) M., Izutsu, K., Takizawa, J., Kusumoto, S., J014. Cheng, Z., Ito. S., Nishio, Na., Xiao, H., Okamoto, M., Yokoyama, M., Takamatsu, Y., Zhang, R., Suzuki, H., Okawa, Y., Murohata, T., Sunami, K., Miyata, A., Murayama, K, Sakai, A., Isobe, K.: Establishment of induced pluripotent Matsumoto, M., Shinagawa, K., Takaki, A., stem cells from aged mice using bone Matsuo, K., Kinoshita, T, Tanimoto, M.: Hepatic marrow-derived myeloid cells. J Mol Cell Biol, 3: toxicity and prognosis in hepatitis C virus-infected 91-98, 2011. (PMID: 21228011) patients with diffuse large B-cell lymphoma treated with rituximab-containing chemotherapy regimens: J015. Chihara, D., Oki, Y., Matsuo, K., Onoda, a Japanese multicenter analysis. Blood, 116: H., Taji, H., Yamamoto, K., Morishima, Y.: 5119-5125, 2010. (PMID: 20823454) Incidence and risk factors for central nervous system relapse in patients with diffuse large B-cell J021. Ennishi, D., Shitara, K., Ito, H., Hosono, lymphoma: analyses with competing risk regression S., Watanabe, M., Ito, S., Sawaki, A., Yatabe, Y., model. Leukemia Lymphoma, 52: 2270-2275, 2011. Yamao, K., Tajima, K., Tanimoto, M., Tanaka, (PMID: 21745170) Hid., Hamajima, N., Matsuo, K.: Association between insulin-like growth factor-1 J016. Chuang, S.C., Agudo, A., Ahrens, W., polymorphisms and stomach cancer risk in a Anatharaman, D., Benhamou, S., Boccia, S., Japanese population. Cancer Sci, 102: 2231-2235, Chen, C., Conway, D., Fabianova, E., Hayes, R.B., 2011. (PMID: 21854509) Healy, C., Holcatova, I., Kjaerheim, K., Lagiou, P., Lazarus, P., Macfarlane, T.V., Mahimkar, M., J022. Ernst, A., Aigner, M., Nakata, S., Engel, Mates, D., Matsuo, K., Merletti, F., Metspalu, A., F., Schlotter, M., Kloor, M., Brand, K., Schmitt, S., Mogenstern, H., Muscat, J., Cadoni, G., Olshan, Steinert, G., Rahbari, N., Koch, M., Radlwimmer, A.F., Purdue, M., Ramroth, H., Rudnai, P., B., Weitz, J., Lichter, P.: A gene signature Schartz, S.M., Simonato, L., Smith, E.M., Sturgis, distinguishing CD133hi from CD133- colorectal E.M., Szeszenia-Dabrowska, N., Talamini, R., cancer cells: essential role for EGR1 and Thomson, P., Wei, Q., Zaidze, D., Zhang, Z.F., downstream factors. Pathology, 43: 220-227, 2011. Znaor, A., Brennan, P., Boffetta, P., Hashibe, M.: (PMID: 21436631) Sequence variants and the risk of head and neck J023. Estecio, M.R., Gallegos, J., Vallot, C., cancer: pooled analysis in the INHANCE Castoro, R.J., Chung, W., Maegawa, S., Oki, Y.,

54 Kondo, Y., Jelinek, J., Shen, L., Hartung, H., Relationship between growth hormone 1 genetic Aplan, P.D., Czerniak, B.A., Liang, S., Issa, J.P.: polymorphism and susceptibility to colorectal Genome architecture marked by retrotransposons cancer. J Hum Genet, 55: 163-166, 2010. (PMID: modulates predisposition to DNA methylation in 20168335) cancer. Genome Res, 10: 1369-1382, 2010. (PMID: J031. Girard, N., Lou, E., Azzoli, C.G., Reddy, 20716667) R., Robson, M., Harlan, M., Orlow, I., Yatabe, Y., J024. Fujii, M., Toyoda, T., Nakanishi, H., Nafa, K., Landanyi, M., Miale, A., Kris, M.G., Yatabe, Y., Sato, A., Matsudaira, Y., Ito, H., Riely, G., Miller, V., Kleing, R., Matsuo, K., Pao, Murakami, H., Kondo, Y., Kondo, E., Hida, T., W.: Analysis of genetic variants in never smokers Tsujimura, T., Osada, H., Sekido, Y.; TGF-b with lung cancer facilitated by an Internet-based synergizes with defects in the Hippo pathway to blood collection protocol: a preliminary report. Clin stimulate human malignant mesothelioma growth. J Cancer Res, 16: 755-763, 2010. (PMID: 20068085) Exp Med, in press. (PMID: 22329991) J032. Goidts, V., Bageritz, J., Puccio, L., Nakata, J025. Fujii, M., Yusa, A., Yokoyama, Y., S., Zapatka, M., Barbus, S., Toedt, G., Campos, B., Kokuryo, T., Tsunoda, N., Oda, K., Nagino, M., Korshunov, A., Momma, S., Van Schaftingen, E., Ishimaru, T., Shimoyama, Y., Utsunomiya, H., Reifenberger, G., Herold-Mende, C., Lichter, P., Iwata, H., Itoh, Y., Itoh, J., Kannagi, R., Radlwimmer, B.: RNAi screening in glioma Kyogashima, M.: Cytoplasmic expression of the stem-like cells identifies PFKFB4 as a key JM403 antigen GlcA-GlcNH3+ on heparan sulfate molecule important for cancer cell survival. glycosaminoglycan in mammary carcinomas - A Oncogene, in press, 2012. (PMID: 22056879) novel proliferative biomarker for breast cancers J033. Goto, Y., Hishida, A., Matsuo, K., Tajima, with high malignancy. Glycoconj J, 27: 661-672, K., Morita, E., Naito, M., Wakai, K., Hamajima, 2010. (PMID: 21046235) N.: PRKCH gene polymorphism is associated with J026. Fujishita, T., Aoki, M., Taketo, M.M.: the risk of severe gastric atrophy. Gastric Cancer, JNK signaling promotes intestinal tumorigenesis 13: 90-94, 2010. (PMID: 20602195) through activation of mTOR complex 1 in Apc 716 J034. Guo, H., Liu, T-Y., Uemura, Y., Jiao. S., mice. Gastroenterology, 140: 1556-1563, 2011. Wang, D., Lin, Z., Narita, Y., Suzuki, M., (PMID: 21320501) Hirosawa, M., Ichihara, Y., Ishihara, O., Kikuchi J027. Fujita, M., Kohanbash, G., Fellows-Mayle, H., Sakamoto, Y., Senju, S., Zhang, Q., Ling, F.: W., Hamilton, R.L., Komohara, Y., Decker, S.A., Bisphenol A in combination with TNFa selectively Ohlfest, J.R., Okada, H.: COX-2 blockade induces Th2 cell-promoting dendritic cells in vitro suppresses gliomagenesis by inhibiting with an estrogen-like activity. Cell Mol Immunol, myeloid-derived suppressor cells. Cancer Res, 71: 7: 227-234, 2010. (PMID: 20383177) 2664-2674, 2011. (PMID: 21324923) J035. Hagiwara, M., Niimi, M., Kawahara, T., J028. Fujita, M., Scheurer, M.E., Decker, S.A., Yamanishi, Y., Nakanishi, H., Arai, F.:On-Chip McDonald, H.A., Kohanbash, G., Kastenhuber, Particle Sorting into Multiple Channels by E.R., Kato, H., Bondy, M.L., Ohlfest, J.R., Okada, Magnetically Driven Microtools. Journal of H.: Role of type-1 interferons in anti-glioma Robotics andMechatronics, 23: 370-377, 2011. immunosurveillance - using mouse studies to guide J036. Hanioka, T., Ojima, M., Tanaka, Hid., examination of novel prognostic markers in humans. Naito, M., Hamajima, N., Matsuse, R.: Intensive Clin Cancer Res, 16: 3409-3419, 2010. (PMID: smoking-cessation intervention in the dental setting. 20472682) J Dent Res, 89: 66-70, 2010. (PMID: 19966043) J029. Fukagawa, T., Sasako, M., Ito, S., J037. Hanioka, T., Ojima, M., Tanaka, K., Nakanishi, H., Iinuma, H., Natsugoe, S., Katai, H., Matsuo, K., Sato, F., Tanaka, Hid.:Causal Shimoda, T.: The prognostic significance of assessment of smoking and tooth loss: A systematic isolated tumor cells in the lymph nodes of gastric review of observational studies. BMC Public Health, cancer patients. Gastric Cancer, 13: 191-196, 2011. 11: 221, 2011. (PMID: 21477320) (PMID: 20820989) J038. Hase, T., Sato, M., Yoshida, K., Girard, L., J030. Gao, C.M., Gong, J.P., Wu, J.Z., Cao, Takeyama, Y., Horio, M., Elshazley, M., Oguri, T., H.X., Ding, J.H., Zhou, J., Liu, Y.T., Li, S.P., Cao, Sekido, Y., Shames, D.S., Gazdar, A.F., Minna, J., Matsuo, K., Takezaki, T., Tajima, K.: J.D., Kondo, M., Hasegawa, Y.: Pivotal role of

55 epithelial cell adhesion molecule in the survival of J045. Hishida, A., Matsuo, K., Goto, Y., Naito, lung cancer cells. Cancer Sci, 102: 1493-1500, M., Wakai, K., Tajima, K., Hamajima, N.:No 2011. (PMID: 21535318) associations of Toll-like receptor 2 (TLR2) -196 to -174del polymorphism with the risk of Helicobacter J039. Hayashi, H., Yamashiro, K., Gotoh, N., pylori seropositivity, gastric atrophy, and gastric Nakanishi, H., Nakata, I., Tsujikawa, A., Otani, A., cancer in Japanese. Gastric Cancer, 13: 251-257, Saito, M., Iida, T., Matsuo, K., Tajima, K., 2010. (PMID: 21128061) Yamada, R., Yoshimura, N.: CFH and ARMS2 variations in age-related macular degeneration, J046. Hishida, A., Matsuo, K., Goto, Y., polypoidal choroidal vasculopathy, and retinal Hamajima, N.: Genetic predisposition to angiomatous proliferation. Invest Ophthalmol Vis Helicobacter pylori-induced gastric precancerous Sci, 51: 5914-1919, 2010. (PMID: 20574013) conditions. World j gastroentero, 2: 369-379, 2010. (PMID: 21160888) J040. Helfand, B.T., Mendez, M.G., Murthy, J047. Hishida, A., Matsuo, K., Goto, Y., Mitsuda, S.N., Shumaker, D.K., Grin, B., Mahammad, S., Y., Hiraki, A., Naito, M., Wakai, K., Kazuo, T., Aebi, U., Wedig, T., Wu, Y.I., Hahn, K.M., Hamajima, N.: No association between AICDA Inagaki, M., Herrmann, H., Goldman, R.D.: 7888 C/T polymorphism, Helicobacter pylori Vimentin organization modulates the formation of seropositivity, and the risk of atrophic gastritis and lamellipodia. Mol Biol Cell, 22: 1274-1289, 2011. gastric cancer in Japanese. Gastric Cancer, 13: (PMID: 21346197) 43-49, 2010. (PMID: 20373075) J041. Hibi, S., Goto, Y., Ando, T., Matsuo, K., J048. Hishida, A., Matsuo, K., Goto, Y., Naito, Wakai, K., Tajima, K., Goto, H., Hamajima, N.: M., Wakai, K., Tajima, K., Hamajima, N.: No association between angiotensin I converting Smoking ehavior and risk of Helicobacter pylori enzyme (ACE) I/D polymorphism and gastric infection, gastric atrophy and gastric cancer in cancer risk among Japanese. Nagoya J Med Sci, 73: Japanese. Asian Pac J Cancer Prev, 11: 313-317, 169-175, 2011. (PMID: 21928698) 2010. (PMID: 20843107) J042. Higashi, M., Kajiyama, H., Shibata, K., J049. Hishida, A., Matsuo, K., Goto, Y., Naito, Mizuno, M., Mizuno, K., Hosono, S., Kawai, M., M., Wakai, K., Tajima, K., Hamajima, N.: Nakanishi, T., Nagasaka, T., Kikkawa, F.: Combined effect of miR-146ars2910164 G/C Survival impact of capsule rupture in stage I clear polymorphism and Toll-like Receptor 4 +3725 G/C cell carcinoma of the ovary in comparison with polymorphism on the risk of severe gastric atrophy other histological types. Gynecol Oncol, 123: in Japanese. Digest Dis Sci, 56: 1131-1137, 2011. 474-478, 2011. (PMID: 21955484) (PMID: 20721625) J043. Hijioka, S., Matsuo, K., Mizuno, N., Hara, J050. Hishida, A., Terazawa, T., Mamiya, T., Ito, K., Mekky, M.A., Vikram, B., Hosoda, W., Yatabe, H., Matsuo, K., Tajima, K., Hamajima, N.: Y., Shimizu, Y., Kondo, S., Tajika, M., Niwa, Y., Efficacy of genotype notification to Japanese Tamada, K., Yamao, K.: Role of endoscopic smokers on smoking cessation - an intervention ultrasound and endoscopic ultrasound-guided study at workplace. Cancer Epidemiol, 34: 96-100, fine-needle aspiration in diagnosing metastasis to 2010. (PMID: 20022836) the pancreas: a tertiary center experience. Pancreatology, 11: 390-398, 2011. (PMID: J051. Hishizawa, M., Kanda, J., Utsunomiya, A., 21894056) Taniguchi, S., Eto, T., Moriuchi, Y., Tanosaki, R., Kawano, F., Miyazaki, Y., Masuda, M., Nagafuji, J044. Hirosawa, T., Torikai, H., Yanagisawa, M., K., Hara, M., Takanashi, M., Kai, S., Atsuta, Y., Kamei, M., Imahashi, N., Demachi-Okamura, A., Suzuki, R., Kawase, T., Matsuo, K., Tanimoto, M., Shiraishi, K., Ito, M., Miyamura, K., Nagamura-Inoue, T., Kato, S., Sakamaki, H., Shibata, K., Kikkawa, F., Morishima, Y., Morishima, Y., Okamura, J., Ichinohe, T., Takahashi, T., Emi, N., Kuzushima, K., Akatsuka, Uchiyama. T.: Transplantation of allogeneic Y.: Mismatched human leukocyte antigen class hematopoietic stem cells for adult T-cell leukemia: II-restricted CD8+ cytotoxic T cells may mediate a nationwide retrospective study. Blood, 116: selective graft-versus-leukemia effects following 1369-1376, 2010. (PMID: 20479287) allogeneic hematopoietic cell transplantation. Cancer Sci, 102: 1281-1286, 2011. (PMID: J052. Horinaka, M., Yoshida, T., Nakata, S., 21466613) Shiraishi, T., Tomosugi, M., Yoshikawa, S., Wakada, M., Sakai, T.: Aclarubicin enhances

56 tumor necrosis factor-related apoptosis-inducing Nutr, 64: 400-409, 2010. (PMID: 20197786) ligand-induced apoptosis through death receptor 5 J060. Hotta, K., Takigawa, N., Matsuo, K., upregulation. Cancer Sci, 103: 282-287, 2012. Kiura, K.: Cure- or Care-Oriented Regimen for (PMID: 22077238) Stage III Non-Small-Cell Lung Cancer? J Clin J053. Horio, M., Sato, M., Takeyama, Y., Oncol, 29: e320, 2011. (PMID: 21402605) Elshazley, M., Yamashita, R., Hase, T., Yoshida, J061. Huang, Q.M., Tomida, S., Masuda, Y., K., Usami, N., Yokoi, K., Sekido, Y., Kondo, M., Arima, C., Cao, K., Kasahara, T.A., Osada, H., Toyokuni, S., Gazdar, A.F., Minna, J.D., Yatabe, Y., Akashi, T., Kamiya, K., Takahashi, T., Hasegawa, Y.: Transient but not stable ZEB1 Suzuki, M.: Regulation of DNA polymerase knockdown dramatically inhibits growth of POLD4 influences genomic instability in lung malignant pleural mesothelioma cells: Implications cancer. Cancer Res, 70: 8707-8417, 2010. (PMID: of gene expression changes resulting from ZEB1 20861182) knockdown. Ann Surg Oncol, in press. (PMID: 22086445) J062. Hwang, J.H., Takagi, M., Murakami, H., Sekido, Y., Shin-Ya, K.: Induction of tubulin J054. Horio, Y., Osada, H., Shimizu, J., Ogawa, polymerization and apoptosis in malignant S., Hida, T., Sekido, Y.: Relationship of mRNA mesothelioma cells by a new compound JBIR-23. expressions of RanBP2 and topoisomerase II Cancer Lett, 300: 189-196, 2011. (PMID: isoforms to cytotoxicity of amrubicin in human 21055871) lung cancer cell lines. Cancer Chemoth Pharm, 66: 237-243, 2010. (PMID: 19809814) J063. Ibi, M., Zou, P., Inoko, A., Shiromizu, T., J055. Hoshino, Y., Nishikawa, K., Ito, Y., Matsuyama, M., Hayashi, Y., Enomoto, M., Mori, Kuzushima, K., Kimura, H.: Kinetics of D., Hirotsune, S., Kiyono, T., Tsukita, S., Goto, H., Epstein-Barr virus load and virus-specific CD8+ T Inagaki, M.: Trichoplein controls microtubule cells in acute infectious mononucleosis. J Clin Virol, anchoring at the centrosome by binding to Odf2 and 50: 244-246, 2011. (PMID: 21185773) ninein. J Cell Sci, 124: 857-864, 2011. (PMID: 21325031) J056. Hosono, S., Kajiyama, H., Mizuno, K., Sakakibara, K., Matsuzawa, K., Takeda, A., Kawai, J064. Ichijima, Y., Yoshioka, K., Yoshioka, Y., M., Nagasaka, T., Kikkawa, F.: Comparison Shinohe, K., Fujimori, H., Unno, J., Takagi, M., between serous and non-serous ovarian cancer as a Goto, H., Inagaki, M., Mizutani, S., Teraoka, H.: prognostic factor in advanced epithelial ovarian DNA lesions induced by replication stress trigger carcinoma after primary debulking surgery. Int J mitotic aberration and tetraploidy development. Clin Oncol, 16: 524-532, 2011. (PMID: 21431342) PLoS ONE, 5: e8821, 2010. (PMID: 20098673) J057. Hosono, S., Kawase, T., Matsuo, K., J065. Igarashi, Y., Kannagi, R.: Watanabe, M., Kajiyama, H., Hirose, K., Suzuki, Glycosphingolipids as mediators of phenotypic T., Kidokoro, K., Ito, H., Nakanishi, T., Yatabe, Y., changes associated with development and cancer Hamajima, N., Kikkawa, F., Tajima, K., Tanaka, progression. J Biochem, 147: 3-8, 2010. (PMID: Hid.: HLA-A alleles and the risk of cervical 20051388) squamous cell carcinoma in Japanese women. J J066. Inaba, T., Ino, K., Kajiyama, H., Shibata, Epidemiol, 20: 295-301, 2010. (PMID: 20501960) K., Yamamoto, E., Kondo, S., Umezu, T., Nawa, A., J058. Hosono, S., Matsuo, K., Hirose, K., Ito, H., Takikawa, O., Kikkawa, F. Indoleamine Suzuki, T., Kawase, T., Watanabe, M., Nakanishi, 2,3-dioxygenase expression predicts impaired T., Tajima, K., Tanaka, Hid.: Weight gain during survival of invasive cervical cancer patients treated adulthood and body weight at age 20 are associated with radical hysterectomy. Gynecol Oncol, 117: with the risk of endometrial cancer in Japanese 423-428, 2010. (PMID: 20350764) women. J Epidemiol, 21: 466-473, 2011. (PMID: 21986192) J067. Inoko, A., Matsuyama, M., Goto, H., Ohmuro-Matsuyama, Y., Hayashi, Y., Enomoto, J059. Hosono, S., Matsuo, K., Kajiyama, H., M., Ibi, M., Urano, T., Yonemura, S., Kiyono, T., Hirose, K., Suzuki, T., Kawase, T., Kidokoro, K., Izawa, I., Inagaki, M.: Trichoplein and Aurora A Nakanishi, T., Hamajima, N., Kikkawa, F., block aberrant primary cilia assembly in Tajima, K., Tanaka, Hid.: Association between proliferating cells. J Cell Biol, in press. dietary calcium and vitamin D intake and cervical carcinogenesis among Japanese women. Eur J Clin J068. Inoue, T., Taguchi, I., Abe, S., Li, G., Hu,

57 R., Nakajima, T., Hara, A., Aoyama, T., Kannagi, applying a propensity score. J Thorac Cardiov Surg, R., Kyogashima, M., Node, K.: Sulfatides are 139: 1001-1006, 2010. (PMID: 19733863) associated with neointimalthickening after vascular J074. Ishiguro, F., Murakami, H., Mizuno, T., injury. Atherosclerosis, 211: 291-296, 2010. Fujii, M., Kondo, Y., Usami, N., Yokoi, K., Osada, (PMID: 20176357) H., Sekido, Y.: Activated leukocyte cell adhesion J069. Inoue, T., Tashiro, Y., Takeuchi, M., molecule (ALCAM) promotes malignant Otani, T., Tsuji-Takayama, K., Okochi, A., Mukae, phenotypes of malignant mesothelioma. J Thorac Y., Koreishi, M., Yamasaki, F., Kumon, H., Oncol, in press. Nakamura, S., Kibata, M., Kondo, E.:Potent J075. Isomura, H., Stinski, M.F., Murata, T., anti-tumor killing activity of the multifunctional Nakayama, S., Chiba, S., Akatsuka, Y., Kanda, T., Treg cell line HOZOT against human tumors with Tsurumi, T.: The human cytomegalovirus UL76 diverse origins. Int J Oncol, 38: 1299-1306, 2011. gene regulates the level of expression of the UL77 (PMID: 21373756) gene. PLoS One, 5: e11901, 2010. (PMID: J070. Iqbal, J., Weisenburger, D.D., Chowdhury, 20689582) A., Tsai, M.Y., Srivastava, G., Greiner, T.C., J076. Isomura, H., Stinski, M.S., Murata, T., Kucuk, C., Deffenbacher, K., Vose, J., Smith, L., Yamashita, Y., Kanda, T., Toyokuni, S., Tsurumi, Au, W.Y., Nakamura, S., Seto, M., Delabie, J., T.: Human cytomegalovirus gene products essential Berger, F., Loong, F., Ko, Y.H., Sng, I., Liu, X., for late viral gene expression assemble into pre- Loughran, T.P., Armitage, J., Chan, W.C.: replication complexes before viral DNA replication. International Peripheral T-cell Lymphoma Project. J Virol, 85: 6629-6644, 2011. (PMID: 21507978) Natural killer cell lymphoma shares strikingly similar molecular features with a group of J077. Ito, H., Matsuo, K., Hosono, S., Watanabe, non-hepatosplenic γδ T-cell lymphoma and is M., Kawase, T., Suzuki, T., Hirai, T., Yatabe, Y., highly sensitive to a novel aurora kinase A inhibitor Tanaka, Hid., Tajima, K.: Association between in vitro. Leukemia, 25: 348-358, 2011. (PMID: CYP7A1 loci and the risk of proximal colorectal 21052088) cancer in Japanese. Int J Mol Epi Genet, 1: 35-46, 2010. (PMID: 21537415) J071. Iqbal, J., Weisenburger, D.D., Greiner, T.C., Vose, J.M., McKeithan, T., Kucuk, C., Geng, J078. Ito, H., Matsuo, K., Tanaka, Hid., Koestler, H., Deffenbacher, K., Smith, L., Dybkaer, K., D.C., Ombao, H., Fulton, J., Shibata, A., Fujita, Nakamura, S., Seto, M., Delabie, J., Berger, F., M., Sugiyama, H., Soda, M., Sobue, T., Mor, V.: Loong, F., Au, W.Y., Ko, Y.H., Sng, I., Armitage, Nonfilter and filter cigarette consumption and the J.O., Chan, W.C.: International Peripheral T-Cell incidence of lung cancer by histological type in Lymphoma Project.: Molecular signatures to Japan and the United States: Analysis of 30-year improve diagnosis in peripheral T-cell lymphoma data from population-based cancer registries. Int J and prognostication in angioimmunoblastic T-cell Cancer, 128: 1918-1928, 2011. (PMID: 20589676) lymphoma. Blood, 115: 1026-1036, 2010. (PMID: J079. Ito, S., Kodera, Y., Mochizuki, Y., Kojima, 19965671) T., Nakanishi, H., Nakao, A.: Phase II clinical trial J072. Ishida, D., Nawa, A., Tanino, T., Goshima, of postoperative S-1 monotherapy for gastric cancer F., Luo, C.H., Iwaki, M., Kajiyama, H., Shibata, patients with Free intraperitoneal cancer cells K., Yamamoto, E., Ino, K., Tsurumi, T., detected by Real-Time RT-PCR. World J surg, 34: Nishiyama, Y., Kikkawa, F.: Enhanced cytotoxicity 2083-2089, 2010. (PMID: 20379713) with a novel system combining the J080. Ito, Y., Nakanishi, H., Kodera, Y., Hirai, paclitaxel-2'-ethylcarbonate prodrug and an HSV T., Nakao, A., Kato, T.: Characterization of a novel amplicon with an attenuated replication-competent lymph node metastasis model from human colonic virus, HF10 as a helper virus. Cancer Lett, 288: cancer and its preclinical use for comparison of 17-27, 2010. (PMID: 19604626) anti-metastatic efficacy between oral S-1 and J073. Ishiguro, F., Matsuo, K., Fukui, T., Mori, UFT/LV. Cancer Sci, 101: 1853-1860, 2010. S., Hatooka, S., Mitsudomi, T.: Effect of selective (PMID: 20500514) lymph node dissection based on patterns of J081. Iwamuro, M., Komaki, T., Kubota, Y., lobe-specific lymph node metastases on patient Seita, M., Kawamoto, H., Yuasa, T., Shahid, J.M., outcome in patients with resectable non-small cell Hassan, R.A., Hassan, W.A., Nakaji, S., lung cancer: a large-scale retrospective cohort study Nishikawa, Y., Kondo, E., Yamamoto, K.,

58 Kobayashi, N.: Comparative analysis of endoderm Prev, 19: 429-434, 2010. (PMID: 20086115) formation efficiency between mouse ES cells and J089. Kanda, J., Matsuo, K., Inoue, M., Iwasaki, iPS cells. Cell Transplant, 19: 831-839, 2010. M., Sawada, N., Shimazu, T., Yamaji, T., Sasazuki, (PMID: 20955658) S., Tsugane, S.: Association of anthropometric J082. Iwamuro, M., Komaki, T., Kubota, Y., characteristics with the risk of malignant lymphoma Seita, M., Kawamoto, H., Yuasa, T., Shahid, J.M., and plasma-cell myeloma in a Japanese population: Hassan, R.A., Hassan, W.A., Nakaji, S., A population-based cohort study. Cancer Epidemiol Nishikawa, Y., Kondo, E., Yamamoto, K., Fox, I.J., Biomarker Prev, 19: 1623-1631, 2010. (PMID: Kobayashi, N.: Hepatic differentiation of mouse 20501768) iPS cells in vitro. Cell Transplant, 19: 841-847, J090. Kanda, J., Matsuo, K., Suzuki, T., Hosono, 2010. (PMID: 20955659) S., Ito, H., Ichinohe, T., Seto, M., Morishima, Y., J083. Iwata, S., Wada, K., Tobita, S., Gotoh, K., Tajima, K., Tanaka, Hid.: Association between Ito, Y., Demachi-Okamura, A., Shimizu, N., obesity and the risk of malignant lymphoma in Nishiyama, Y., Kimura, H.: Quantitative analysis Japanese: a case-control study. Int J Cancer, 126: of Epstein-Barr virus (EBV)-related gene 2416-2425, 2010. (PMID: 19821488) expression in patients with chronic active EBV J091. Kanda, T., Shibata, S., Saito, S., Murata, infection. J Gen Virol, 91: 42-50, 2010. (PMID: T., Isomura, H., Yoshiyama, H., Takada, K., 19793909) Tsurumi, T.: Unexpected instability of Family of J084. Ju, H.X., An, B., Okamoto, Y., Shinjo, K., Repeats (FR), the critical cis-acting sequence Kanemitsu, Y., Komori, K., Hirai, T., Shimizu, Y., required for EBV latent infection, in EBV-BAC Sano, T., Sawaki, A., Tajika, M., Yamao, K., Fujii, systems. PLoS One, 6: e27758, 2011. (PMID: M., Murakami, H., Osada, H., Ito, H., Takeuchi, I., 22114684) Sekido, Y., Kondo, Y.: Distinct profiles of J092. Kannagi, R., Sakuma, K., Miyazaki, K., epigenetic evolution between colorectal cancers Lim, K.T., Yusa, A., Yin, J., Izawa, M.: Altered with and without metastasis. Am J Pathol, 178: expression of glycan genes in cancers induced by 1835-1846, 2011. (PMID: 21406167) epigenetic silencing and tumor hypoxia: clues in the J085. Kajiyama, H., Shibata, K., Mizuno, M., ongoing search for new tumor markers. Cancer Sci, Hosono, S., Kawai, M., Nagasaka, T., Kikkawa, 101: 586-593, 2010. (PMID: 20085584) F.: Fertility-sparing surgery in patients with J093. Karube, K., Nakagawa, M., Tsuzuki, S., clear-cell carcinoma of the ovary: Is it possible? Takeuchi, I., Honma, K., Nakashima, Y., Shimizu, Hum Reprod, 26: 3297-3302, 2011. (PMID: N., Ko, Y.H., Morishima, Y., Ohshima, K., 22016417) Nakamura, S., Seto M.: Identification of FOXO3 J086. Kakizaki, F., Aoki, K., Miyoshi, H., and PRDM1 as tumor suppressor gene candidates in Carrasco, N., Aoki, M., Taketo, M.M.: CDX NK cell neoplasms by genomic and functional transcription factors positively regulate expression analyses. Blood, 118: 3195-3204, 2011. (PMID: of Solute Carrier Family 5, Member 8 in the colonic 21690554) epithelium. Gastroenterology, 138: 627-635, 2010. J094. Kasahara, K., Goto, H., Enomoto, M., (PMID: 19900445) Tomono, Y., Kiyono, T., Inagaki, M.: J087. Kamiyama, S., Ichimiya, T., Ikehara, Y., 14-3-3gamma mediates Cdc25A proteolysis to Takase, T., Fujimoto, I., Suda, T., Nakamori, S., block premature mitotic entry after DNA damage. Nakamura, M., Nakayama, F., Irimura, T., EMBO J, 29: 2802-2812, 2010. (PMID: 20639859) Nakanishi, H., Watanabe, M., Narimatsu, H., J095. Katayama, T., Matsuo, K., Kosaka, T., Nishihara, S.: Expression and the role of Sueda, T., Yatabe, Y., Mitsudomi, T.: Effect of 3'-phosphoadenosine 5'-phosphosulfate transporters gefitinib on the survival of patients with recurrence in human colorectal carcinoma. Glycobiology, 21: of lung adenocarcinoma after surgery: A 235-346, 2011. (PMID: 20978009) retrospective case-matching cohort study. J088. Kanda, J., Matsuo, K., Inoue, M., Iwasak, Surg Oncol, 19: e144-9, 2010. (PMID: 20705455) M., Sawada, N., Shimazu, T., Yamaji, T., Sasazuki, J096. Kato, H., Kagami, Y, Nakagawa, M., S., Tsugane, S.: Association of alcohol intake with Karnan, S, Yatabe, Y., Morishima, Y., Seto, M.: the risk of malignant lymphoma and plasma-cell IL-4/CD40L Co-Stimulation Induces Long-Term myeloma in Japanese: A population-based cohort Proliferation for CD10-Positive Germinal Center B study (JPHC study). Cancer Epidemiol Biomarker

59 Cell-Like Diffuse Large B-CeIl Lymphoma. The (PMID: 19798093) Open Leukemia Journal, 3: 60-68, 2010. J104. Kimura, Y., Sato, K., Arakawa, F., Karube, J097. Kato, H., Kagami, Y., Kodaira, T., Oka, S., K., Nomura, Y., Shimizu, K., Aoki, R., Hashikawa, Oki, Y., Chihara, D., Taji, H., Yatabe, Y., K., Yoshida, S., Kiyasu, J., Takeuchi, M., Nino, D., Nakamura, T., Nakamura, S., Seto, M., Yamamoto, Sugita, Y., Morito, T., Yoshino, T., Nakamura, S., K., Morishima, Y.: Nodal relapse after helicobacter Kikuchi, M., Ohshima, K.: Mantle cell lymphoma pylori eradication in a patient with primary shows three morphological evolutions of classical, localized gastric mucosa-associated lymphoid tissue intermediate, and aggressive forms, which occur in lymphoma. Am J Gastroenterol, 106: 549-551, parallel with increased labeling index of cyclin D1 2011. (PMID: 21378774) and Ki-67. Cancer Sci, 101: 806-814, 2010. (PMID: 20002441) J098. Kato, H., Yamamoto, K., Matsuo, K., Oki, Y., Taji, H., Kuwatsuka, Y., Seto, M., Kagami, Y., J105. Kishida, Y., Natsume, A., Kondo, Y., Morishima, Y.: Clinical impact and predisposing Takeuchi, I., An, B., Okamoto, Y., Shinjo, K., factors of delayed-onset neutropenia after Saito, K., Ando, H., Ohka, F., Sekido, Y., autologous hematopoietic stem-cell transplantation Wakabayashi, T.: Epigenetic subclassification of for B-cell non-Hodgkin lymphoma: association meningiomas based on genome-wide DNA with an incremental risk of infectious events. Ann methylation analyses. Carcinogenesis, 33: 436-441, Oncol, 21: 1699-1705, 2010. (PMID: 20172906) 2011. (PMID: 22102699) J099. Kato, H., Yamamoto, K., Taji, H., Oki, Y., J106. Kishida, Y., Natsume, A., Toda, H., Toi, Y., Chihara, D., Seto, M., Kagami, Y., Morishima, Y.: Motomura, K., Koyama, H., Matsuda, K., Interstitial pneumonia after autologous Nakayama, O., Sato, M., Suzuki, M., Kondo, Y., hematopoietic stem cell transplantation in B-cell Wakabayashi, T.: Correlation between quantified non-hodgkin lymphoma. Clin Lymphoma Myeloma promoter methylation and enzymatic activity of O Leuk, 11: 483-489, 2011. (PMID: 21978956) (6)-methylguanine-DNA methyltransferase in glioblastomas. Tumour Biol, in press. (PMID: J100. Kawamoto, H., Yuasa, T., Kubota, Y., 22274924) Seita, M., Sasamoto, H., Javed, S.M., Hayashi, T., Nakahara, H., Hassan, R., Iwamuro, M., Kondo, J107. Kitamura, T., Fujishita, T., Loetscher, P., E., Nakaji, S., Tanaka, N., Kobayashi, N.: Revesz, L., Hashida, H., Kizaka-Kondoh, S., Aoki, Characteristics of CD133(+) human colon cancer M, Taketo, M.M.: Inactivation of chemokine (C-C SW620 cells. Cell Transplant, 19: 857-864, 2010. motif) receptor 1 (CCR1) suppresses colon cancer (PMID: 20587144) liver metastasis by blocking accumulation of immature myeloid cells in a mouse model. Proc J101. Kawase, T., Matsuo, K., Suzuki, T., Hirose, Natl Acad Sci USA, 107: 13063-13068, 2010. K., Hosono, S., Watanabe, M., Inagaki, M., Iwata, (PMID: 20616008) H., Tanaka, Hid., Tajima, K.: Association between vitamin D and calcium intake and breast cancer risk J108. Kitawaki, T., Kadowaki, N., Fukunaga, K., according to menopausal status and receptor status Kasai, Y., Maekawa, T., Ohmori, K., Itoh, T., in Japan. Cancer Sci, 101: 1234-1240, 2010. Shimizu, A., Kuzushima, K., Kondo, T., Ishikawa, (PMID: 20151981) T., Uchiyama, T.: Cross-priming of CD8(+) T cells in vivo by dendritic cells pulsed with autologous J102. Kimura, T., Kohno, H., Matsuoka, Y., apoptotic leukemic cells in immunotherapy for Murakami, M., Nakatsuka, R., Hase, M., Yasuda, elderly patients with acute myeloid leukemia. Exp K., Uemura, Y., Sasaki, Y., Fukunaga, S., Sonoda, Hematol, 39: 424-433, 2011. (PMID: 21216276) Y.: CXCL8 enhances the angiogenic activity of umbilical cord blood-derived outgrowth endothelial J109. Kitawaki, T., Kadowaki, N., Fukunaga, K., cells in vitro. Cell Biol Int, 35: 201-208, 2011. Kasai, Y., Maekawa, T., Ohmori, K., Kondo, T., (PMID: 20958269) Maekawa, R., Takahara, M., Nieda, M., Kuzushima, K., Ishikawa, T., Uchiyama, T.: A J103. Kimura, T., Matsuoka, Y., Murakami, M., phase I/IIa clinical trial of immunotherapy for Kimura, T., Takahashi, M., Nakamoto, T., Yasuda, elderly patients with acute myeloid leukaemia using K., Matsui, K., Kobayashi, K., Imai, S., Asano, H., dendritic cells co-pulsed with WT1 peptide and Nakatsuka, R., Uemura, Y., Sasaki, Y., Sonoda, zoledronate. Brit J Haematol, 153: 796-799, 2011. Y.: In vivo dynamics of human cord blood-derived (PMID: 21477159) CD34- SCID-repopulating cells using intra-bone marrow injection. Leukemia, 24: 162-168, 2010. J110. Ko, Y.H., Karnan, S., Kim, K.M., Park,

60 C.K., Kang, E.S., Kim, Y.H., Kang, W.K., Kim, Huang, C.X., Hou, M.F., Shen, H., Hu, Z., Wang, S.J., Kim, W.S., Lee, W.Y., Chun, H.K., Seto, M.: F., Deming, S.L., Kelley, M.C., Shrubsole, M.J., Enteropathy-associated T-cell lymphoma- A Khoo, U.S., Chan, K.Y.K., Chan, S.Y., Haima, clinicopathologic and array comparative genomic C.A., Henderson, B.E., Le, Marchand, L., Iwasaki, hybridization study. Hum Pathol, 41: 1231-1237, M., Kasuga, Y., Tsugane, S., Matsuo, K., Tajima, 2010. (PMID: 20399483) K., Iwata, H., Huang, B., Shi, J., Li, G., Wen, W., Gao, Y.T., Lu, W., Zheng, W.: Identification of a J111. Kobayashi, Y., Niwa, Y., Tajika, M., Kawai, functional genetic variant at 16q12.1 for breast H., Kondo, S., Hara, K., Mizuno, N., Hijioka, S., cancer risk: Results from the Asia Breast Cancer Sawaki, A., Matsuo, K., Nakagawa, H., Nakamura, Consortium. Plos Genet, 6: e1001002, 2010. Y., Yamao, K.: Serum tumor antigen REG4 as a (PMID: 20585626) useful diagnostic biomarker in gastric cancer. Hepato-gastroenterol, 57: 1631-1634, 2010. (PMID: J118. Maeda, O., Shibata, K., Hosono, S., 21443133) Fujiwara, S., Kajiyama, H., Ino, K., Nawa, A., Tamakoshi, K., Kikkawa, F.: Spectrin αII and βII J112. Konishi, K., Watanabe, Y., Shen, L., Guo, Y., Castoro, R.J., Kondo, K., Chung, W., Ahmed, tetramers contribute to platinum anticancer drug S., Jelinek, J., Boumber, Y.A., Estecio, M.R., resistance in ovarian serous adenocarcinoma. Int J Maegawa, S., Kondo, Y., Itoh, F., Imawari, M., Cancer, 130: 113-121, 2012. (PMID: 21328338) Hamilton, S.R., Issa, J.P.: DNA methylation J119. Maseki, S., Ijichi, K., Tanaka, Ha., Fujii, profiles of primary colorectal carcinoma and M., Hasegawa, Y., Ogawa T, Murakami, S., matched liver metastasis. PLoS One,6: e27889, Kondo, E., Nakanishi, H.: Acquisition of EMT 2011. (PMID: 22132162) phenotype in the gefitinib-resistant cells of a head J113. Kumar, V., Matsuo, K., Takahashi, A., and neck squamous cell carcinoma cell line through Hosono, N., Tsunoda, T., Kamatani, N., Kong, Akt/GSK-3β/snail signaling pathway. Br J Cancer, S.Y., Nakagawa, H., Cui, R., Tanikawa, C., Seto, in press. (PMID: 22315058) M., Morishima, Y., Kubo, M., Nakamura, Y., J120. Matsui, M., Shimizu, Y., Ikehara, Y., Matsuda, K.: Common variants on 14q32 and Kondo, E., Kodera, Y., Nakanishi, H.: Targeted 13q12 are associated with DLBCL susceptibility. J delivery of oligomannose-coated liposome to the Hum Genet, 56: 436-439, 2011. (PMID: 21471979) omental micrometastasis by peritoneal J114. Kuroda, A., Tsukamoto, Y., Nguyen, L.T., macrophages from patients with gastric cancer. Noguchi, T., Takeuchi, I., Uchida, M., Uchida, T., Cancer Sci, 101: 1670-1677, 2010. (PMID: Hijiya, N., Nakada, C., Okimoto, T., Kodama, M., 20507320) Murakami, K., Matsuura, K., Seto, M., Ito, H., J121. Matsuoka, Y., Sasaki, Y., Nakatsuka, R., Fujioka, T., Moriyama, M.: Genomic profiling of Takahashi, M., Iwaki, R., Uemura, Y., Sonoda, Y.: submucosal-invasive gastric cancer by array-based The low level of c-kit expression marks deeply comparative genomic hybridization. PLoS One, 6: quiescent murine hematopoietic stem cells. Stem e22313, 2011. (PMID: 21811585) Cells, 29: 1783-1791, 2011. (PMID: 21898688) J115. Li, P., Goto, H., Kasahara, K., Matsuyama, J122. Matsuse, M., Takahashi, M., Mitsutake, M., Wang, Z., Yatabe, Y., Kiyono, T. and Inagaki, N., Nishihara, E., Hirokawa, M., Kawaguchi, T., M.: P90 RSK arranges Chk1 in the nucleus for Rogounovitch, T., Saenko, V., Bychkov, A., Suzuki, monitoring of genomic integrity during cell K., Matsuo, K., Tajima, K., Miyauchi, A., Yamada, proliferation. Mol Biol Cell, in press. (PMID: R., Matsuda, F., Yamashita, S.: The FOXE1 and 22357623) NKX2-1 loci are associated with susceptibility to J116. Lin, Y., Yagyu, K., Egawa, N., Ueno, M., papillary thyroid carcinoma in the Japanese Mori, M., Nakao, H., Ishii, H., Nakamura, K., population. J Med Genet, 48: 645-648, 2011. Wakai, K., Hosono, S., Tamakoshi, A., Kikuchi, (PMID: 21730105) S.: An overview of genetic polymorphisms and J123. Matsuyama, M., Goto, H., Kasahara, K., pancreatic cancer risk in molecular epidemiologic Kawakami, Y., Nakanishi, M., Kiyono, T., studies. J Epidemiol, 21: 2-12, 2011. (PMID: Goshima, N., Inagaki, M.: Nuclear Chk1 prevents 21071884) premature mitotic entry. J Cell Sci, 124: 2113-2119, J117. Long, J., Cai, Q., Shu, X.O., Qu, S., Li, C., 2011. (PMID: 21628425) Zheng, Y., Gu, K., Wang, W., Xiang, Y.B., Cheng, J124. Miyata, T., Yonekura, K., Utsunomiya, A., J., Chen, K., Zhang, L., Zheng, H., Shen, C.Y.,

61 Kanekura, T., Nakamura, S., Seto, M.: Cutaneous 19903244) type adult T-cell leukemia/lymphoma is a J131. Muramatsu, H., Kojima, S., Yoshimi, A., characteristic subtype and includes erythema/papule Atsuta, Y., Kato, K., Nagatoshi, Y., Inoue, M., and nodule/tumor subgroups. Int J Cancer, 126: Koike, K., Kawase, T., Ito, M., Kurosawa, H., 1521-1528, 2010. (PMID: 19739121) Tanizawa, A., Tono, C., Hamamoto, K., Hotta, N., J125. Mizuno, T., Murakami, H., Fujii, M., Watanabe, A., Morishima, Y., Kawa, K., Shimada, Ishiguro, F., Tanaka, I., Kondo, Y., Akatsuka, S., H.: Outcome of 125 children with CML who Toyokuni, S., Yokoi, K., Osada, H., Sekido, Y.: received transplants from unrelated donors: the YAP induces malignant mesothelioma cell Japan Marrow Donor Program (JMDP). Biol Blood proliferation by upregulating transcription of cell Marrow Tr, 16: 231-238, 2010. (PMID: 19800016) cycle promoting genes. Oncogene, in press. (PMID: J132. Muramatsu, H., Makishima, H., 22286761) Jankowska, A.M., Cazzolli, H., O'Keefe, C., J126. Mizuta, S., Matsuo, K., Yagasaki, F., Yoshida, N., Xu, Y., Nishio, No., Hama, A., Yujiri, T., Hatta, Y., Kimura, Y., Ueda, Y., Yagasaki, H., Takahashi, Y., Kato, K., Manabe, A., Kanamori, H., Usui, N., Akiyama, H., Miyazaki, Kojima, S., Maciejewski, J.P.: Mutations of an E3 Y., Ohtake, S., Atsuta, Y., Sakamaki, H., Kawa, K., ubiquitin ligase c-Cbl but not TET2 mutations are Morishima, Y., Ohnishi, K., Naoe, T., Ohno, R.: pathogenic in juvenile myelomonocytic leukemia. Pre-transplantation Imatinib-based therapy Blood, 115: 1969-1975, 2010. (PMID: 20008299) improves the clinical outcomes of allogeneic J133. Muramatsu, H., Takahashi, Y., Sakaguchi, hematopoietic stem cell transplantation in patients H., Shimada, A., Nishio, No., Hama, A., Doisaki, with newly diagnosed BCR-ABL-positive acute S., Yagasaki, H., Matsumoto, K., Kato, K., Kojima, lymphoblastic leukemia: a comparative study. S.: Excellent outcomes of children with CML Leukemia, 25: 41-47, 2011. (PMID: 20944676) treated with imatinib mesylate compared to that in J127. Motomura, K., Natsume, A., Kishida, Y., pre-imatinib era. Int J Hematol, 93: 186-191, 2011. Higashi, H., Kondo, Y., Nakasu, Y., Abe, T., (PMID: 21234820) Namba, H., Wakai, K., Wakabayashi, T.: Benefits J134. Muramatsu, H., Takahashi, Y., of interferon-β and temozolomide combination Shimoyama, Y., Doisaki, S., Nishio, No., Ito, Y., therapy for newly diagnosed primary glioblastoma Hama, A., Shimada, A., Yagasaki, H., Ito, M., with the unmethylated MGMT promoter: a Kojima, S.: CD20-negative Epstein-Barr multicenter study. Cancer, 117: 1721-1730, 2011. virus-associated post-transplant lymphoproliferative (PMID: 21061328) disease refractory to rituximab in a patient with J128. Murakami, H., Mizuno, T., Taniguchi, T., severe aplastic anemia. Int J Hematol, 93: 779-781, Fujii, M., Ishiguro, F., Fukui, T., Akatsuka, S., 2011. (PMID: 21559814) Horio, Y., Hida, T., Kondo, Y., Toyokuni, S., J135. Murata, T., Hotta, N., Toyama, S., Osada, H., Sekido, Y.: LATS2 is a tumor Nakayama, S., Chiba, S., Isomura, H., Ohshima, suppressor gene of malignant mesothelioma. T., Kanda, T., Tsurumi, T.: Transcriptional Cancer Res, 71: 873-883, 2011. (PMID: 21245096) repression by sumoylation of Epstein-Barr virus J129. Murakami, H., Nakanishi, H., Tanaka, BZLF1 protein correlates with association of Ha., Ito, S., Misawa, K., Ito, Y., Ikehara, Y., histone deacetylase. J Biol Chem, 285: Kondo, E., Kodera, Y.: Establishment and 23925-23935, 2010. (PMID: 20516063) characterization of novel gastric signet-ring cell and J136. Murata, T., Kondo, Y., Sugimoto, A., non signet-ring cell, poorly-differentiated Kawashima, D., Saito, S., Isomura, H., Kanda, T., adenocarcinoma cell lines with low and high Tsurumi, T.: Epigenetic histone modification of malignant potential. Gastric Cancer, in press. Epstein-Barr virus BZLF1 promoter during latency J130. Murakami, M., Ito, H., Hagiwara, K., and reactivation in Raji cells. J Virol, in press. Yoshida, K., Sobue, S., Ichihara, M., Takagi, A., J137. Murata, T., Noda, C., Saito, S., Kojima, T., Tanaka, K., Tamiya-Koizumi, K., Kawashima, D., Sugimoto, A., Isomura, H., Kanda, Kyogashima, M., Suzuki, M., Banno, M., Nozawa, T., Yokoyama, K., Tsurumi, T.: Involvement of Jun Y., Murate, T.: ATRA inhibits ceramide kinase dimerization protein 2 (JDP2) in the maintenance of transcription in a human neuroblastoma cell line, Epstein-Barr virus latency. J Biol Chem, 286: SH-SY5Y cells: the role of COUP-TFI. J 22007-22016, 2011. (PMID: 21525011) Neurochem, 112: 511-520, 2010. (PMID:

62 J138. Nagafuji, K., Matsuo, K., Teshima, T., J145. Nakao, M., Hosono, S., Ito, H., Watanabe, Mori, S., Sakamaki, H., Hidaka, M., Ogawa, H., M., Mizuno, N., Yatabe, Y., Yamao, K., Ueda, R., Kodera, Y., Kanda, Y., Maruta, A., Mori, T., Tajima, K., Tanaka, Hid., Matsuo, K.: Interaction Yoshida, F., Ichinohe, T., Kasai, M., Takatsuka, between IGF-1 polymorphisms and overweight for Y., Kubo, K., Sao, H., Atsuta, Y., Suzuki, R., the risk of pancreatic cancer in Japanese. Int J Mol Yoshida, T., Tsuchida, M., Harada, M.: Peripheral Epidemiol Genet, 2: 354-366, 2011. (PMID: blood stem cell versus bone marrow transplantation 22199998) from HLA-identical sibling donors in patietns with J146. Nakao, M., Matsuo, K., Hosono, S., Ogata, leukemia: A propensity score-based comparison S., Ito, H., Watanabe, M., Mizuno, N., Iida, S., from the Japanese Society of Hematopoietic Stem Satoh, S., Yatabe, Y., Yamao, K., Ueda, R., Tajima, Cell Transplantation Registry. Int J Hematol, 91: K., Tanaka, Hid.: ABO blood group alleles and the 855-864, 2010. (PMID: 20464644) risk of pancreatic cancer in Japanese population. J139. Nagai, R., Hashimoto, R., Tanaka, Y., Cancer Sci, 102: 1076-1080, 2011. (PMID: Taguchi, O., Sato, M., Matsukage, A., Yamaguchi, 21306478) M.: Syntrophin-2 is required for eye development J147. Nakao, M., Matsuo, K., Ito, H., Shitara, K., in Drosophila. Exp Cell Res, 316: 272-285, 2010. Hosono, S., Watanabe, M., Ito, S., Sawaki, A., Iida, (PMID: 19836389) S., Sato, S., Yatabe, Y., Yamao, K., Ueda, R., J140. Nagashio, R., Arai, E., Ojima, H., Kosuge, Tajima, K., Hamajima, N., Tanaka, Hid.:ABO T., Kondo, Y., Kanai, Y.: Carcinogenetic risk Genotype and the Risk of Gastric Cancer, Atrophic estimation based on quantification of DNA Gastritis, and Helicobacter pylori Infection. Cancer methylation levels in liver tissue at the Epidemiol Biomarkers Prev, 20: 1665-1672, 2011. precancerous stage. Int J Cancer, 129: 1170-1179, (PMID: 21680535) 2011. (PMID: 21400512) J148. Nakata, I., Yamashiro, K., Yamada, R., J141. Nakada, C., Tsukamoto, Y., Matsuura, K., Gotoh, N., Nakanishi, H., Hayashi, H., Tsujikawa, Nguyen, TL., Hijiya, N., Uchida, T., Sato, F., A., Otani, A., Saito, M., Iida, T., Oishi, A., Matsuo, Mimata, H., Seto, M., Moriyama, M.: K., Tajima, K., Matsuda, F., Yoshimura, N.: Overexpression of miR-210, a downstream target of Association between the SERPING1 gene and HIF1α, causes centrosome amplification in renal age-related macular degeneration and polypoidal carcinoma cells. J Pathol, 224: 280-288, 2011. choroidal vasculopathy in Japanese. PLoS One, 6: (PMID: 21465485) e19108, 2011. (PMID: 21526158) J142. Nakagawa, M., Tsuzuki, S., Honma, K., J149. Nakatsuka, R., Nozaki T., Uemura, Y., Taguchi, O., Seto, M.: Synergistic effect of Bcl2, Matsuoka, Y., Sasaki Y., Shinohara, M., Ohura, Myc and Ccnd1 transforms mouse primary B-cells K., Sonoda, Y.: 5-Aza-2’-deoxycytidine treatment into malignant cells. Haematologica, 96: 1318-1326, induces skeletal myogenic differentiation of mouse 2011. (PMID: 21606168) dental pulp stem cells. Arch Oral Biol, 55: 350-357, 2010. (PMID: 20362276) J143. Nakanishi, H., Hayamashi, H., Yamada, R., Yamashiro, K., Nakata, I., Shimada, N., J150. Nakaya, T., Kuwahara, K., Ohta, K., Ohno-Matsui, K., Mochizuki, M., Ozaki, M., Kitabatake, M., Toda, T., Takeda, N., Tani, T., Yoshitake, S., Kuriyama, S., Saito, M., Iida, T., Kondo, E., Sakaguchi, N.: Critical role of Pcid2 in Matsuo, K., Matsuda, F., Yoshimura, N.: Single B cell survival through the regulation of MAD2 nucleotide polymorphisms in promoter region of expression. J Immunol, 185: 5180-5187, 2010. matrix metalloproteinase 1, 2, and 3 in Japanese (PMID: 20870947) with High Myopia. Invest Ophthalmol Vis Sci, 51: J151. Nakayama, S., Murata, T., Yasui, Y., 4432-4436, 2010. (PMID: 20435584) Murayama, K., Isomura, H., Kanda, T., Tsurumi, J144. Nakanishi, H., Yamashiro, K., Yamada, R., T.: Tetrameric ring formation of Epstein-Barr virus Gotoh, N., Hayashi, H., Nakata, I., Saito, M., Iida, polymerase processivity factor is crucial for viral T., Oishi, A., Kurimoto, Y., Matsuo, K., Tajima, K., replication. J Virol, 84: 12589-12598, 2010. Matsuda, F., Yoshimura, N.: Joint effect of (PMID: 20926567) cigarette smoking and CFH and J152. Narita, A., Muramatsu, H., Takahashi, Y., LOC387715/HTRA1 polymorphisms on polypoidal Sakaguchi, H., Doisaki, S., Nishio, No., Hama, A., choroidal vasculopathy. Invest Ophthalmol Vis Sci, Shimada, A., Ito, M., Kojima, S.:. 51: 6183-6187, 2010. (PMID: 20688737) Autoimmune-like hepatitis following unrelated

63 BMT successfully treated with rituximab. Bone dyskeratosis congenita. Pediatr Transplant, 15: Marrow Transplant, in publication, 2011. (PMID: 161-166, 2011. (PMID: 21176016) 21666737) J159. Nishio, No., Takahashi, Y., Tanaka, M., J153. Navarro-Alvarez, N., Kondo, E., Xu, Y., Yoshida, N., Sakaguchi, H., Doisaki, S., Soto-gutierrez, A., Kawamoto, H., Hassan, W., Hama, A., Muramatsu, H., Shimada, A., Kojima, Yuasa, T., Kubota, Y., Seita, M., Nakahara, H., S.: Aberrant phosphorylation of STAT5 by Hayashi, T., Nishikawa, Y., Hassan, R.A., Javed, granulocyte0macrophage colony-stimulating factor S.M., Noguchi, H., Matsumoto, S., Nakaji, S., in infant cytomegalovirus infection mimicking Tanaka, N., Kobayashi, N.: Isolation and juvenile myelomonocytic leukemia. Leuk Res, 35: propagation of a human CD133-negative colon 1261-1264, 2011. (PMID: 21571368) tumor-derived cell line withtumorigenic and J160. Noda, C., Murata, T., Kanda, T., angiogenic properties. Cell Transplant, 19: 865-877, Yoshiyama, H., Sugimoto, A., Kawashima, D., 2010. (PMID: 20587145) Saito, S., Isomura, H., Tsurumi, T.: Identification J154. Navarro-Alvarez, N., Soto-Gutierrez, A., and characterization of CCAAT enhancer-binding Chen, Y., Caballero-Corbalan, J., Hassan, W., protein (C/EBP) as a transcriptional activator for Kobayashi, S., Takei, J., Kondo, Y., Iwamuro, M., Epstein-Barr virus oncogene latent membrane Yamamoto, K., Kondo, E., Tanaka, N., FoxI, J., protein 1. J Biol Chem, 286: 42524-42533, 2011. Kobayashi, N.: Intramuscular transplantation of (PMID: 22013073) engineered hepatictissueconstructs corrects acute J161. Ochi, T., Fujiwara, H., Okamoto, S., An, and chronic liver failure in mice. J Hepatol, 52: J., Nagai, K., Shirakata, T., Mineno, J., 211-219, 2010. (PMID: 20022655) Kuzushima, K., Shiku, H., Yasukawa, M.: Novel J155. Nicolotti, N., Chuang, S.C., Cadoni, G., adoptive T-cell immunotherapy using a Arzani, D., Petrelli, L., Bosetti, C., Brenner, H., WT1-specific TCR vector encoding silencers for Hosono, S., La, Vecchia, C., Matsuo, K., Muller, endogenous TCRs shows marked antileukemia H., Muscat, J., Paludetti, G., Ricciardi, G., reactivity and safety. Blood, 118:1495-1503, 2011. Boffetta, P., Hashibe, M., Boccia, S.: Recreational (PMID: 21673345) physical activity and risk of head and neck cancer: a J162. Ohka, F., Natsume, A., Motomura, K., pooled analysis within the international head and Kishida, Y., Kondo, Y., Abe, T., Nakasu, Y., neck cancer epidemiology (INHANCE) Consortium. Namba, H., Wakai, K., Fukui, T., Momota, H., Eur J Epidemiol, 26: 619-628, 2011. (PMID: Iwami, K., Kinjo, S., Ito, M., Fujii, M., 21842237) Wakabayashi, T.: The global DNA methylation J156. Niibe, Y., Watanabe, J., Tsunoda, S., Arai, surrogate LINE-1 methylation is correlated with M., Arai, T., Kawaguchi, M., Ono, S., Numata, A., MGMT promoter methylation and is a better Matsuo, K., Jobo, T., Unno, N., Hayakawa, K.: prognostic factor for glioma. PLoS One,6: Concomitant expression of HER2 and HIF-1alpha e23332,2011. (PMID: 21829728) is a predictor of poor prognosis in uterine cervical J163. Ohmura, K., Terao, C., Maruya, E., carcinoma treated with concurrent Katayama, M., Matoba, K., Shimada, K., chemoradiotherapy: prospective analysis Murasawa, A., Honjo, S., Takasugi, K., Tohma, S., (KGROG0501). Eur J Gynaecol Oncol, 31: Matsuo, K., Tajima, K., Yukawa, N., Kawabata, D., 491-496, 2010. (PMID: 21061787) Nojima, T., Fujii, T., Yamada, R., Saji, H., J157. Nishikawa, E., Osada, H., Okazaki, Y., Matsuda, F., Mimori, T.: Anti-citrullinated peptide Arima, C., Tomida, S., Tatematsu, Y., Taguchi, A., antibody-negative RA is a genetically distinct Shimada, Y., Yanagisawa, K., Yatabe, Y., subset: a definitive study using only bone-erosive Toyokuni, S., Sekido, Y., Takahashi, T.: miR-375 ACPA-negative rheumatoid arthritis. is activated by ASH1 and inhibits YAP1 in a Rheumatology, 49: 2298-2304, 2010. (PMID: lineage-dependent manner in lung cancer. Cancer 20833643) Res, 71: 6165-6173, 2011. (PMID: 21856745) J164. Okamoto, N., Kuwahara, K., Ohta, K., J158. Nishio, No., Takahashi, Y., Ohashi, H., Kitabatake, M., Takagi, K., Mizuta, H., Kondo, E., Doisaki, S., Muramatsu, H., Hama, A., Shimada, Sakaguchi, N.: Germinal center-associated nuclear A., Yagasaki, H., Kojima, S.: Reduced-intensity protein (GANP) is involved in mRNA export of conditioning for alternative donor hematopoietic Shugoshin-1 required for centromere cohesion and stem cell transplantation in patients with in sister-chromatid exchange. Genes Cells, 15:

64 471-484, 2010. (PMID: 20384790) Y.W., Tanaka, Hid., Tajima, K., Hirose, H., Tanioka, F., Matsuno, Y., Sugimura, H., Kato, S., J165. Okamoto, Y., Sawaki, A., Ito, S., Nishida, Nakamura, T., Nishina, T., Yasui, W., Aoyagi, K., T., Takahashi, T., Toyota, M., Suzuki, H., Sasaki, H., Yanagihara, K., Katai, H., Shimoda, T., Shinomura, Y., Takeuchi, I., Shinjo, K., An, B., Yoshida, T., Nakamura, Y., Hirohashi, S., Ito, H., Yamao, K., Fujii, M., Murakami, H., Sakamoto, H.: A functional single nucleotide Osada, H., Kataoka, H., Joh, T., Sekido, Y., Kondo, polymorphism in mucin 1, at chromosome 1q22, Y.: Aberrant DNA methylation associated with determines susceptibility to diffuse-type gastric aggressiveness of gastrointestinal stromal tumour. cancer. Gastroenterology, 140: 892-902, 2011. GUT, 61: 392-401, 2012. (PMID: 21708825) (PMID: 21070779) J166. Okasaka, T., Usami, N., Taniguchi, T., J172. Saito, K., Kondo, E., Matsushita, M.: Kawaguchi, K., Okagawa, T., Suzuki, H., Matsuo, MicroRNA 130 family regulates the hypoxia K., Yokoi, K.: Can non-performance of radical response signal through the P-body protein DDX6. systematic mediastinal lymphadenectomy be Nucleic Acids Res, 39: 6086-6099, 2011. (PMID: justified in elderly lung cancer patients? An 21486751) evaluation using propensity-based survival analysis. Eur J Cardiothorac Surg, 38: 27-33, 2010. (PMID: J173. Sasaki, K., Kohanbash, G., Hoji, A., Ueda, 20189406) R., McDonald, H.A., Reihart, T.A., Martinson, J., Lotze, M.T., Marincola, F.M., Wand, E., Fujita, J167. Ozaki, H., Matsuzaki, H., Ando, H., Kaji, M., Okada, H.: miR-17-92 expression in H., Nakanishi, H., Ikehara, Y., Narimatsu, H.: differentiated T cells - implications for cancer Enhancement of metastatic ability by ectopic immunotherapy. J Transl Med, 8: 17, 2010. (PMID: expressionof ST6GalNAcI on a gastric cancer cell 20167088) line in a mouse model. Clin Exp Metastasis, in press. (PMID: 22228572) J174. Sasazuki, S., Inoue, M., Tsuji, I., Sugawara, Y., Tamakoshi, A., Matsuo, K., Wakai, J168. Oze, I., Matsuo, K., Hosono, S., Ito, H., K., Nagata, C., Tanaka, K., Mizoue, T., Tsugane, Kawase, T., Watanabe, M., Suzuki, T., Hatooka, S., S.: Body mass index and mortality from all causes Yatabe, Y., Hasegawa, Y., Shinoda, M., Tajima, K., and major causes in japanese: results of a pooled Tanaka, Hid.: Comparison between Self-report analysis of 7 large-scale cohort studies. J Epidemiol, facial flushing after alcohol consumption and 21: 417-430, 2011. (PMID: 21908941) ALDH2 Glu504Lys Polymorphism for risk of upper Aerodigestive Tract Cancer in a Japanese J175. Sato, F., Oze, I., Kawakita, D., Yamamoto, Population. Cancer Sci, 101: 1875-1880, 2010. N., Ito, H., Hosono, S., Suzuki, T., Kawase, T., (PMID: 20518787) Furue, H., Watanabe, M., Hatooka, S., Yatabe, Y., Hasegawa, Y., Shinoda, M., Ueda, M., Tajima, K., J169. Park, J.Y., Matsuo, K., Suzuki, T., Ito, H., Tanaka, Hid., Matsuo, K.: Inverse association Hosono, S., Kawase, T., Watanabe, M., Oze, I., between toothbrushing and upper aerodigestive Hida, T., Yatabe, Y., Mitsudomi, T., Takezaki, T., tract cancer risk in a japanese population. Head Tajima, K., Tanaka, Hid.: Impact of smoking on Neck, 33: 1628-1637, 2011. (PMID: 21259377) lung cancer risk is stronger in those with the homozygous aldehyde dehydrogenase 2 null allele J176. Sato, K., Misawa, N., Nie, C., Satou, Y., in a Japanese population. Carcinogenesis, 31: Iwakiri, D., Matsuoka, M., Takahashi, R., 660-665, 2010. (PMID: 20093384) Kuzushima, K., Ito, M., Takada, K., Koyanagi, Y.: A novel animal model of Epstein-Barr J170. Renault-Mihara, F., Katoh, H., Ikegami, virus-associated hemophagocytic T., Iwanami, A., Mukaino, M., Yasuda, A., Nori, lymphohistiocytosis in humanized mice. Blood, S., Mabuchi, Y., Tada, H., Shibata, S., Saito, K., 117: 5663-5673, 2011. (PMID: 21467545) Matsushita, M., Kaibuchi, K., Okada, S., Toyama, Y., Nakamura, M., Okano, H.: Beneficial J177. Sato, Y., Shirata, N., Murata, T., Nakasu, compaction of spinal cord lesion by migrating S., Kudoh, A., Iwahori, S., Nakayama, S., Chiba, astrocytes through glycogen synthase kinase-3 S., Isomura, H., Kanda, T., Tsurumi, T.: Transient inhibition. EMBO Mol Med, 3: 682-696, 2011. increases in p53-responsible gene expression at (PMID: 21898827) early stages of Epstein-Barr virus productive replication. Cell Cycle, 9: 807-814, 2010. (PMID: J171. Saeki, N., Saito, A., Choi, I.J., Matsuo, K., 20139729) Ohnami, S., Totsuka, H., Chiku, S., Kuchiba, A., Lee, Y.S., Yoon, K.A., Kook, M.C., Park, S.R., Kim, J178. Segawa, Y., Kiura, K., Hotta, K., Takigawa,

65 N., Tabata, M., Matsuo, K., Yoshioka, H., Hayashi, molecular targeting agents and curative surgery. H., Kawai, H., Aoe, K., Maeda, T., Ueoka, H., Oncology, 81: 167-174, 2011. (PMID: 22057124) Tanimoto, M.: A randomized phase II study of a J184. Shitara, K., Matsuo, K., Mizota, A., Kondo, combination of docetaxel and S-1 versus docetaxel C., Nomura, M., Takahari, D., Yokota, T., Ura, T., monotherapy in patients with non-small-cell lung Ito, S., Tajika, M., Muro, K.: Association of cancer previously treated with platinum-based fluoropyrimidines, platinum agents, taxanes, and chemotherapy: Results of Okayama Lung Cancer irinotecan in any line of chemotherapy with Study Group (OLCSG) trial 0503. J Thorac Oncol, survival in patients with advanced gastric cancer. 5: 1430-1444, 2010. (PMID: 20651615) Gastric Cancer, 14: 155-160, 2011. (PMID: J179. Segawa, Y., Kiura, K., Takigawa, N., 21340668) Kamei, H., Harita, S., Hiraki, S., Watanabe, Y., J185. Shitara, K., Matsuo, K., Oze, I., Mizota, A., Sugimoto, K., Shibayama, T., Yonei, T., Ueoka, H., Kondo, C., Nomura, M., Yokota, T., Takahari, D., Takemoto, M., Kanazawa, S., Takata, I., Nogami, Ura, T., Muro, K.: Meta-analysis of neutropenia or N., Hotta, K., Hiraki, A., Tabata, M., Matsuo, K., leucopenia as a prognostic factor in patients with Tanimoto, M.: A randomized phase III trial malignant disease undergoing chemotherapy. comparing docetaxel and cisplatin combination Cancer chemoth pharm, 68: 301-307, 2011. (PMID: chemotherapy to mitomycin, vindesine and 20960191) cisplatin combination chemotherapy with concurrent thoracic radiotherapy in locally J186. Shitara, K., Matsuo, K., Takahari, D., advanced non-small-cell lung cancer: OLCSG 0007. Yokota, T., Shibata, T., Ura, T., Ito, S., Sawaki, A., J Clin Oncol, 28: 3299-3306, 2010. (PMID: Tajika, M., Kawai, H., Muro, K.: Neutropenia as a 20530281) prognostic factor in advanced gastric cancer patients undergoing second-line chemotherapy with J180. Shen, L., Kantarjian, H., Guo, Y., Lin, E., weekly paclitaxel. Ann Oncol, 21: 2403-2409, 2010. Shan, J., Huang, X., Berry, D., Ahmed, S., Zhu, (PMID: 20494962) W., Pierce, S., Kondo, Y., Oki, Y., Jelinek, J., Hussain, S., Estey, E., Issa, J.P.J.: DNA J187. Shitara, K., Muro, K., Ito, S., Sawaki, A., methylation predicts survival and response to Tajika, M., Kawai, H., Yokota, T., Takahari, D., therapy in patients with myelodysplastic syndromes. Shibata, T., Ura, T., Ito, H., Hosono, S., Kawase, J Clin Oncol, 28: 605-613, 2010. (PMID: T., Watanabe, M., Tajima, K., Yatabe, Y., Tanaka, 20038729) Hid., Matsuo, K.: Folate intake along with genetic polymorphisms in methylenetetrahydrofolate J181. Shitara, K., Matsuo, K., Hatooka, S., Ura, reductase and thymidylate synthase in patients with T., Takahari, D., Yokota, T., Abe, T., Kawai, H., advanced gastric cancer. Cancer Epidemiol Tajika, M., Kodaira, T., Shinoda, M., Tajima, K., Biomarker Prev, 19: 1311-1319, 2010. (PMID: Muro, K., Tanaka, Hid.: Heavy smoking history 20447923) interacts with chemoradiotherapy for esophageal cancer prognosis: a retrospective study. Cancer Sci, J188. Shitara, K., Oze, I., Mizota, A., Kondo, C., 101: 1001-1006, 2010. (PMID: 20085588) Nomura, M., Yokota, T., Takahari, D., Ura, T., Yuki, S., Komatsu, Y., Matsuo, K., Muro, K.: J182. Shitara, K., Matsuo, K., Ito, S., Sawaki, A., Randomized phase II study comparing dose Kawai, H., Yokota, T., Takahari, D., Shibata, T., escalated weekly paclitaxel vs. standard dose Ura, T., Ito, H., Hosono, S., Kawase, T., Watanabe, weekly paclitaxel for patients with previously M., Tajima, K., Yatabe, Y., Tanaka, Hid., Muro, treated advanced gastric cancer. Jpn J Clin Oncol, K.: Effects of genetic polymorphisms in the 41: 287-290, 2011. (PMID: 21068025) ABCB1 gene on clinical outcomes in patients with gastric cancer treated by second-line chemotherapy. J189. Shitara, K., Ura, T., Matsuo, K., Takahari, Asian Pac J Cancer Prev, 11: 447-452, 2010. D., Yokota, T., Yuki, S., Yoshida, M., Utsunomiya, (PMID: 20843132) S., Sato, Y., Yamaura, H., Kato, M., Inaba, Y., Tajika, M., Kawai, H., Yamazaki, K., Komatsu, Y., J183. Shitara, K., Matsuo, K., Kondo, C., Muro, K.: Sensitivity to previous irinotecan Takahari, D., Ura, T., Inaba, Y., Yamamura, H., treatment does not predict the efficacy of Sato, Y., Kato, M., Kanemitsu, Y., Sano, T., combination chemotherapy with cetuximab plus Shimizu, Y., Muro, K.: Prolonged survival of irinotecan for wild-type KRAS metastatic colorectal patients with metastatic colorectal cancer following cancer. Eur J Cancer, 47: 2673-2680, 2011. (PMID: first-line oxaliplatin-based chemotherapy with 21652203)

66 J190. Sobue, T., Inoue, M., Tanaka, Hid., and system lymphoma. Blood, 117: 1291-1300, 2011. 46 Members.: Cancer Registry and Epidemiological (PMID: 21088137) Study Working Group report. Jpn J Clin Oncol, J198. Tajima, K., Ohashi, R., Sekido, Y., Hida, 40(Suppl 1): i76-i81, 2010. (PMID: 20870925) T., Nara, T., Hashimoto, M., Iwakami, S., J191. Sonoshita, M., Aoki, M., Fuwa, H., Aoki, Minakata, K., Yae, T., Tahahashi, F., Saya, H., K., Hosogi, H., Sakai, Y., Hashida, H., Takahashi, K.: Osteopontin mediated enhanced Takabayashi, A., Sasaki, M., Robine, S., Itoh, K., hyaluronan binding induces multidrug resistance in Yoshioka, K., Kakizaki, F., Kitamura, T., Oshima. mesothelioma cells. Oncogene, 29: 1941-1951, M., Taketo, M.M.: Suppression of colon cancer 2010. (PMID: 20101228) metastasis by Aes through inhibition of Notch J199. Tajiri, H., Tanaka, Hid., Brooks, S., signaling. Cancer Cell, 19: 125-137, 2011. (PMID: Takano, T.: Reduction of hepatocellular carcinoma 21251616) in childhood after introduction of selective J192. Suda, K., Murakami, I., Katayama, T., vaccination against hepatitis B virus for infants Tomizawa, K., Osada, H., Sekido, Y., Maehara, Y., born to HBV carrier mothers. Cancer Cause Control, Yatabe, Y., Mitsudomi, T.: Reciprocal and 22: 523-527, 2011. (PMID: 21191808) complementary role of MET amplification and J200. Takayama, R., Nakagawa, H., Sawaki, A., EGFR T790M mutation in acquired resistance to Mizuno, N., Kawai, H., Tajika, M., Yatabe, Y., kinase inhibitors in lung cancer. Clin Cancer Res, Matsuo, K., Uehara, R., Ono, K., Nakamura, Y., 16: 5489-5498, 2010. (PMID: 21062933) Yamao, K.: Serum tumor antigen REG4 as a J193. Suda, K., Tomizawa, K., Fujii, M., diagnostic biomarker in pancreatic ductal Murakami, H., Osada, H., Maehara, Y., Yatabe, Y., adenocarcinoma. J Gastroenterol, 45: 52-59, 2010. Sekido, Y., Mitsudomi, T.: Epithelial to (PMID: 19789838) mesenchymal transition in an EGFR-mutant lung J201. Takemura, Y., Satoh, M., Satoh, K., cancer cell line with acquired resistance to erlotinib. Hamada, H., Sekido, Y., Kubota, S.: High dose of J Thorac Oncol, 6: 1152-1161, 2011. (PMID: ascorbic acid induces cell death in mesothelioma 21597390) cells. Biochem Biophys Res Commun, 394: J194. Suda, K., Tomizawa, K., Osada, H., 249-253, 2010. (PMID: 20171954) Maehara, Y., Yatabe, Y., Sekido, Y., Mitsudomi, J202. Takeyama, Y., Sato, M., Horio, M., Hase, T.: Conversion from the "oncogene addiction" to T., Yoshida, K., Yokoyama, T., Nakashima, H., "drug addiction" by intensive inhibition of the Hashimoto, N., Sekido, Y., Gazdar, A.F., Minna, EGFR and MET in lung cancer with activating J.D., Kondo, M., Hasegawa, Y.: Knockdown of EGFR mutation. Lung Cancer, in press. (PMID: ZEB1, a master epithelial-to-mesenchymal 22133747) transition (EMT) gene suppresses J195. Sugimoto, A., Kanda, T., Yamashita, Y., anchorage-independent cell growth of lung cancer Murata, T., Saito, S., Kawashima, D., Isomura, H., cells. Cancer Lett, 296: 216-224, 2010. (PMID: Nishiyama, Y., Tsurumi, T.: Spatiotemporally 20452118) different DNA repair systems participate during J203. Takigawa, N., Kiura, K., Segawa, Y., Epstein-Barr virus genome maturation. J Virol, 85: Hotta, K., Tamaoki, A., Touda, Y., Nagata, T., 6127-6135, 2011. (PMID: 21490093) Watanabe, K., Gemba, K., Moritaka, T., Horita, N., J196. Sugimoto, Y., Muramatsu, H., Makishima, Takeda, H., Okimoto, N., Takemoto, M., Matsuo, H., Prince, C., Jankowska, A.M., Yoshida, N., Xu, K., Shinkai, T., Tabata, M., Ueoka, H., Knazawa, Y., Nishio, No., Hama, A., Yagasaki, H., S., Tanimoto, M.: Benefits and adverse events Takahashi, Y., Kato, K., Manabe, A., Kojima, S., among elderly patients receiving concurrent Maciejewski, J.P.: Spectrum of molecular defects chemoradiotherapy for locally advanced non-small in juvenile myelomonocytic leukaemia includes cell lung cancer: Analysis of the Okayama Lung ASXL1 mutations. Brit J Haematol, 150: 83-87, Cancer Study Group Trial 0007. J Thorac Oncol, 6: 2010. (PMID: 20408841) 1087-1091, 2011. (PMID: 21415777) J197. Sung, C.O., Kim, S.C., Karnan, S., Karube, J204. Tamura, U., Tanaka, T., Okamura, T., K., Shin, H.J., Nam, D.H., Suh, Y.L., Kim, S.H., Kadowaki, T., Yamato, H., Tanaka, Hid., Kim, J.Y., Kim, S.J., Kim, W.S., Seto, M., Ko, Nakamura, M., Okayama, A., Ueshima, H., Y.H.: Genomic profiling combined with gene Yamagata, Z., for the HIPOP-OHP research expression profiling in primary central nervous group.: Changes in weight, cardiovascular risk

67 factors and estimated risk for coronary heart disease der Heijden, H.F., Kiemeney, L.A., Isla, D., following smoking cessation in Japanese male Mayordomo, J.I., Rafnar, T., Stefansson, K., workers: HIPOP-OHP Study. J Atheroscler Thromb, Zhang, Z.F., Chang, S.C., Kim, J.H., Hong, Y.C., 17: 12-20, 2010. (PMID: 20081325) Duell, E.J., Andrew, A.S., Lejbkowicz, F., Rennert, G., Muller, H., Brenner, H., Le, Marchand. L., J205. Tanaka, Hid., Tsukuma, H., Oshima, A.: Benhamou, S., Bouchardy, C., Teare, M.D., Xue, Long-term prospective study of 6104 survivors of X., McLaughlin, J., Liu, G., McKay, J.D., srsenic poisoning during infancy due to Brennan, P., Spitz, M.R.: Replication of lung contaminated milk powder in1955. J Epidemiol, 20: cancer susceptibility loci at chromosomes 15q25, 439-445, 2010. (PMID: 20736507) 5p15, and 6p21: a pooled analysis from the J206. Tanaka, K., Yamada, M., Tamiya-Koizumi, International Lung Cancer Consortium. J Natl K., Kannagi, R., Aoyama, T., Hara, A., Cancer I, 102: 959-971, 2010. (PMID: 20548021) Kyogashima, M.: Systematic analyses of free J212. Truong, T., Sauter, W., McKay, J.D., ceramide species and ceramide species comprising Hosgood, H.D., Gallagher, C., Amos, C.I., Spitz, neutral glycosphingolipids by MALDI-TOF MS M., Muscat, J., Lazarus, P., Illig, T., Wichmann, with high-energy CID. Glycoconj J, 28: 67-87, E., Bickeboller, H., Risch, A., Dienemann, H., 2011. (PMID: 21400001) Zhang, Z.F., Naeim, B.P., Yang, P., Zienolddiny, J207. Tanaka, M., Tanaka, Hid., Tsukuma, H., S., Haugen, A., Le, Marchand, L., Hong, Y.C., Ioka, A., Oshima, A., Nakahara, T.: Risk factors Kim, J.H., Duell, E.J., Andrew, A.S., Kiyohara, C., for intrahepatic cholangiocarcinoma: a possible role Shen, H., Matsuo, K., Suzuki, T., Seow, A., Ng, of hepatitis B virus. J Viral Hepatitis, 17: 742-748, D.P., Lan, Q., Zaridze, D., Szeszenia-dabrowska, 2010. (PMID: 20002305) N., Lissowska, J., Rudnai, P., Fabianova, E., J208. Taniguchi, C., Hibino, F., Kawaguchi, E., Constantinescu, V., Bencko, V., Foretova, L., Janout, V., Caporaso, N.E., Albanes, D., Thun, M., Maruguchi, M., Tokunaga, N., Saka, H., Oze, I., Ito, H., Hiraki, A., Nakamura, S., Tanaka, Hid.: Landi, M.T., Trubicka, J., Lener, M., Lubinski, J., Perceptions and practices of Japanese nurses EPIC-lung, Wang, Y., Chabrier, A., Boffetta, P., regarding tobacco intervention for cancer patients. J Brenann, P., Hung, R.J.: International Lung Cancer Consortium: Coordinated genotyping of 10 Epidemiol, 21: 391-397, 2011. (PMID: 21821967) potential lung cancer susceptibility variants. J209. Terao, C., Ohmura, K., Kochi, Y., Ikari, K., Carcinogenesis, 31: 625-633, 2010. (PMID: Maruya, E., Katayama, M., Shimada, K., 20106900) Murasawa, A., Honjo, S., Takasugi, K., Matsuo, K., Tajima, K., Suzuki, A., Yamamoto, K., J213. Tsukamoto, Y., Nakada, C., Noguchi, T., Momohara, S., Yamanaka, H., Yamada, R., Saji, Tanigawa, M., Nguyen, L.T., Uchida, T., Hijiya, N., Matsuura, K., Fujioka, T., Seto, M., Moriyama, H., Matsuda, F., Mimori, T.: A large-scale association study identified multiple HLA-DRB1 M.: MicroRNA-375 is downregulated in gastric alleles associated with ACPA-negative rheumatoid carcinomas and regulates cell survival by targeting arthritis in Japanese subjects. Ann Rheum Dis, 70: PDK1 and 14-3-3zeta. Cancer Res, 70: 2339-2349, 2010. (PMID: 20215506) 2134-2139, 2011. (PMID: 21873689) J210. Tomizawa, K., Suda, K., Onozato, R., J214. Tsuzuki, S., Seto, M.: Expansion of Kosaka, T., Endoh, H., Sekido, Y., Shigematsu, H., functionally defined mouse hematopoietic stem and progenitor cells by a short isoform of Kuwano, H., Yatabe, Y., Mitsudomi, T.: Prognostic and predictive implications of HER2/ERBB2/neu RUNX1/AML1. Blood, 119: 727-735, 2012. gene mutations in lung cancers. Lung Cancer, 74: (PMID: 22130803) 139-144, 2011. (PMID: 21353324) J215. Tsuzuki, S., Taguchi, O., Seto, M.: Promotion and maintenance of leukemia by ERG. J211. Truong, T., Hung, R.J., Amos, C.I., Wu, X., Bickeboller, H., Rosenberger, A., Sauter, W., Blood, 117: 3758-3868, 2011. (PMID: 21321361) Illig, T., Wichmann, H.E., Risch, A., Dienemann, J216. Uchida, M., Tsukamoto, Y., Uchida, T., H., Kaaks, R., Yang, P., Jiang, R., Wiencke, J.K., Ishikawa, Y., Nagai, T., Hijiya, N., Nguyen, L T., Wrensch, M., Hansen, H., Kelsey, K.T., Matsuo, Nakada, C., Kuroda, A., Okimoto, T., Kodama, M., K., Tajima, K., Schwartz, A.G., Wenzlaff, A., Seow, Murakami, K., Noguchi, T., Matsuura, K., A., Ying, C., Staratschek-Jox, A., Nurnberg, P., Tanigawa, M., Seto, M., Ito, H., Fujioka, T., Stoelben, E., Wolf, J., Lazarus, P., Muscat, J.E., Takeuchi, I., Moriyama, M.: Genomic profiling of Gallagher, C.J., Zienolddiny, S., Haugen, A., van

68 gastric carcinoma in situ and adenomas by Research Group for the Development and array-based comparative genomic hybridization. J Evaluation of Cancer Prevention Strategies in Pathol, 221: 96-105, 2010. (PMID: 20217874) Japan.: Lung cancer risk and consumption of vegetables and fruit: an evaluation based on a J217. Ueda, M., Shimada, T., Goto, Y., Tei, K., systematic review of epidemiological evidence Nakai, S., Hisa, Y., Kannagi, R.: Expression of from Japan. Jpn J Clin Oncol, 41: 693-708, 2011. CC-chemokine receptor 7 (CCR7) and (PMID: 21422002) CXC-chemokine receptor 4 (CXCR4) in head and neck squamous cell carcinoma. Auris Nasus Larynx, J222. Warren, E.H., Fujii, N., Akatsuka, Y., 37: 488-495, 2010. (PMID: 20036791) Chaney, C.N., Mito, J.K., Loeb, K.R., Gooley, T.A., Brown, M.L., Koo, K.K, Rosinski, K.V., Ogawa, S., J218. Umino, A., Nakagawa, M., Utsunomiya, Matsubara, A., Appelbaum, F.R., Riddell, S.R.: A., Tsukasaki, K., Taira, N., Katayama, N., Seto, Therapy of relapsed leukemia after allogeneic M.: Clonal evolution of adult T-cell hematopoietic cell transplant with T cells specific leukemia/lymphoma takes place in lymph node. for minor histocompatibility antigens. Blood, 115: Blood, 117: 5473-5478, 2011. (PMID: 21447829) 3869-3878, 2010. (PMID: 20124217) J219. Villalobos, I.B., Takahashi, Y., Akatsuka, J223. Yagasaki, H., Takahashi, Y., Hama, A., Y., Muramatsu, H., Nishio, No., Hama, A., Kudo, K., Nishio, No., Muramatsu, H., Tanaka, Yagasaki, H., Saji, H., Kato, M., Ogawa, S., M., Yoshida, N., Matsumoto, K., Watanabe, N., Kojima, S.: Relapse of leukemia with loss of Kato, K., Horibe, K., Kojima, S.: Comparison of mismatched HLA resulting from uniparental matched-sibling donor BMT and unrelated donor disomy after haploidentical hematopoietic stem cell BMT in children and adolescent with acquired transplantation. Blood, 115: 3158-3161, 2010. severe aplastic anemia. Bone marrow transpl, 45: (PMID: 20124217) 1508-1513, 2010. (PMID: 20118992) J220. Wakai, K., Hamajima, N., Okada, R., J224. Yamada, T., Matsumoto, K., Wang, W., Li, Naito, M., Morita, E., Hishida, A., Kawai, S., Q., Nishioka, Y., Sekido, Y., Sone, S., Yano, S.: Nishio, K., Yin, G., Asai, Y., Matsuo, K., Hosono, Hepatocyte growth factor reduces susceptibility to S., Ito, H., Watanabe, M., Kawase, T., Suzuki, T., an irreversible epidermal growth factor receptor Tajima, K., Tanaka, K., Higaki, Y., Hara, M., inhibitor in EGFR-T790M mutant lung cancer. Clin Imaizumi, T., Taguchi, N., Nakamura, K., Nanri, Cancer Res, 16:174-183, 2010. (PMID: 20008840) H., Sakamoto, T., Horita, M., Shinchi, K., Kita, Y., Turin, T.C., Rumana, N., Matsui, K., Miura, K., J225. Yamaguchi, T., Yanagisawa, K., Sugiyama, Ueshima, H., Takashima, N., Nakamura, Y., R., Hosono, Y., Shimada, Y., Arima, C., Kato, S., Suzuki, S., Ando, R., Hosono, A., Imaeda, N., Tomida, S., Suzuki, M., Osada, H., Takahashi, T.: Shibata, K., Goto, C., Hattori, N., Fukatsu, M., NKX2-1/TITF1/TTF-1-induced ROR1 is required Yamada, T., Tokudome, S., Takezaki, T., Niimura, to sustain EGFR survival signaling in lung H., Hirasada, K., Nakamura, A., Tatebo, M., adenocarcinoma. Cancer Cell, 21:348-361, 2012. Ogawa, S., Tsunematsu, N., Chiba, S., Mikami, H., (PMID: 22439932) Kono, S., Ohnaka, K., Takayanagi, R., Watanabe, J226. Yamahira, A., Narita, M., Nakamura, T., Y., Ozaki, E., Shigeta, M., Kuriyama, N., Watanabe, N., Kaji, M., Taniguchi, T., Hashimoto, Yoshikawa, A., Matsui, D., Watanabe, I., Inoue, S., Furukawa, T., Toba, K., Aizawa, Y., K., Ozasa, K., Mitani, S., Arisawa, K., Uemura, H., Kuzushima, K., Takahashi, M.: Generation of Hiyoshi, M., Takami, H., Yamaguchi, M., antigen-specific cytotoxic T lymphocytes using a Nakamoto, M., Takeda, H., Kubo, M., Tanaka, leukemic plasmacytoid dendritic cell line as antigen Hid.: Profile of participants and genotype presenting cells. Leukemia Res, 35: 793-799, 2011. distributions of 108 polymorphisms in a (PMID: 21216464) cross-sectional study of associations of genotypes with lifestyle and clinical factors: a project in the J227. Yamamoto, M., Kondo, E., Takeuchi, M., japan multi-institutional collaborative cohort Harashima, A., Otani, T., Tsuji-Takayama, K., (J-MICC) study. J Epidemiol, 21: 223-235, 2011. Yamasaki, F., Kumon, H., Kibata, M., Nakamura, (PMID: 21467728) S.: miR-155, a Modulator of FOXO3a Protein Expression, Is Underexpressed and Cannot Be J221. Wakai, K., Matsuo, K., Nagata, C., Mizoue, Upregulated by Stimulation of HOZOT, a Line of T., Tanaka, K., Tsuji, I., Sasazuki, S., Shimazu, T., Multifunctional Treg. PLoS One, 6: e16841, 2011. Sawada, N., Inoue, M., Tsugane, S.; for the (PMID: 21304824)

69 J228. Yamamoto, T., Navarro-Alvarez, N., Zhang, X., Inoue, M., Matsuo, K., He, J., Chandra, Soto-Gutierrez, A., Yuasa, T., Iwamuro, M., Gupta, P., Rmadas, K., Tsugane, S., Irie, F., Kubota, Y., Seita, M., Kawamoto, H., Javed, S.M., Tamakoshi, A., Gao, Y.T., Wang, R., Shu, X.O., Kondo, E., Noguchi, H., Kobayashi, S., Nakaji, S., Tsuji, I., Kuriyama, S., Tanaka, Hid., Satoh, H., Kobayashi, N.: Treatment of acute liver failure in Chen, C.J., Yuan, J.M., Yoo, K.Y., Ahsan, H., Pan, mice by hepatocyte xenotransplantation. Cell W.H., Gu, D., Suryankant, Pednekar, M., Transplant, 19: 799-806, 2010. (PMID: 20573299) Sauvaget, C.K., Sasazuki, S., Sairenchi, T., Yang, G., Xiang, Y.B., Nagai, M., Suzuki, T., Nishino, Y., J229. Yanagisawa, K., Konishi, H., Arima, C., You, S.L., Koh, W.P., Park, S., Chen, Y., Shen, Tomida, S., Takeuchi, T., Shimada, Y., Yatabe, Y., C.Y., Thornquist, M., Feng, Z., Kang, D.H., Mitsudomi, T., Osada, H., Takahashi, T.: Novel Boffetta, P., Potter, J.: Association between metastasis-related gene CIM functions in the body-mass index and risk of death in more than 1 regulation of multiple cellular stress-response million Asians. New Engl J Med, 364: 719-729, pathways. Cancer Res, 70: 9949-9958, 2010. 2011. (PMID: 21345101) (PMID: 21118962) J237. Zhu, X., Fujita, M., Snyder, L.A., Okada, J230. Yasuda, A., Noguchi, K., Minoshima, M., H.: Systemic delivery of neutralizing antibody Kashiwazaki, G., Kanda, T., Katayama, K., targeting CCL2 for glioma therapy. J Neurooncol, Mitsuhashi, J., Bando, T., Sugiyama, H., 104: 83-92, 2011. (PMID: 21116835) Sugimoto, Y.: DNA ligand designed to antagonize EBNA1 represses Epstein-Barr virus-induced J238. Zhu, X., Junecko, B.F., Fujita, M., Ueda, immortalization. Cancer Sci, 102: 2221-2230, 2011. R., Kohanbash, G., Kasternhuber, E.R., (PMID: 21910783) McDonald, H.A., Liu, Y., Kalinski, P., Reinhart, T.A., Salazar, A.M., Okada, H.: Poly-ICLC J231. Yatabe, Y., Matsuo, K., Mitsudomi, T.: promotes the efficacy of peptide vaccines by Heterogeneous distribution of EGFR mutations is inducing CXCL10 in intracranial gliomas in IFN-α extremely rare in lung adenocarcinoma. J Clin γ Oncol, 29: 2972-2977, 2011. (PMID: 21730270) and IFN- dependent manners. Cancer Immunol Immunother, 59: 1401-1409, 2010. (PMID: J232. Yoshida, K., Yatabe, Y., Park, J., Ogawa, 20549206) S., Park, J.Y., Shimizu, J., Horio, Y., Matsuo, K., J239. Katanoda, K., Sobue, T., Satoh, H., Mitsudomi, T., Hida, T.: Clinical outcomes of Tajima, K., Suzuki, T., Nakatsuka, H., Takezaki, advanced non-small cell lung cancer patients T., Nakayama, T., Nitta, H., Tanabe, K., screened for epidermal growth factor receptor gene Tominaga, S.: An association between long-term mutations. J Cancer Res Clin Oncol, 136: 527-535, exposure to ambient air pollution and mortality 2010. (PMID: 19777258) from lung cancer and respiratory diseases in Japan. J233. Yusa, A., Miyazaki, K., Kimura, N., Izawa, J Epidemiol, 21: 132-143, 2011. (PMID: 21325732) M., Kannagi, R.: Epigenetic Silencing of the sulfate J240. Li, M., Zhou, Y., Chen, P., Yang, H., Yuan, transporter gene DTDST induces sialyl lewisx X., Tajima, K., Cao, J., Wang, H.: Genetic variants expression and accelerates proliferation of colon on chromosome 8q24 and colorectal cancer risk: A cancer cells. Cancer Res, 70: 4064-4073, 2010. case-control study in China and meta-analysis of (PMID: 20460514) the published literature. Plos One, 6 (3): e18251, 2011. (PMID: 21455501) J234. Zhang, R., Ito. S., Nishio, Na., Cheng, Z., J241. Katanoda, K., Saika, K., Yamamoto, S., Suzuki, H., Isobe, K.: Up-regulation of Tanaka, S., Oshima, A., Nakamura, M., Satoh, H., Gr1+CD11b+ population in spleen of dextran Tajima, K., Suzuki, T., Tamakoshi, A., Tsugane, sulfate sodium administered mice works to repair S., Sobue, T.: Projected cancer mortality among colitis. Inflamm Allergy Drug targets, 10: 39-46, Japanese males under different smoking prevalence 2011. (PMID: 21184649) scenorios: evidence for tobacco control goal setting. J235. Zhang, R., Ito. S., Nishio, Na., Cheng, Z., Jpn J Clin Oncol, 41: 483-489, 2011. (PMID: Suzuki, H., Isobe, K.: Dextran sulphate sodium 21262873) increases splenic Gr1(+)CD11b(+) cells which J242. Liu, Y.T., Gao, C.M., Ding, J.H., Li, S.P., accelerate recovery from coltis following Cao, H.X., Wu, J.Z., Tang, J.H., Qian, Y., Tajima, intravenous transplantation. Clin Exp Immunol, K.: Physiological, reproductive factors and breast 164: 417-427, 2011. (PMID: 21413942) cancer risk in Jiangsu Province of China. Asia Pacific J Cancer Prev, 12: 787-790, 2011. (PMID: J236. Zheng, W., MacLerran, D.F., Rolland, B., 21627384)

70 J243. Jiang, A.R., Gao, C.M., Ding, J.H., Li, in Jiangsu province of china. Asian Pac J Cancer S.P., Liu, Y.T., Cao, H.X., Wu, J.Z., Tang, J.H., Prev, 13: 33-35, 2012. (PMID: 22502696) Qian, Y., Tajima, K.: Abortions and breast cancer risk in premenopausal and postmenopausal women

71 419-424, 2010. (PMID: 20882440) Reviews and Books R010. Okamoto, Y., Kondo, Y.: General Mechanisms of Cancers - Genetic and epigenetic R001. Fujita, M., Okada, H.: alterations in inflammation-related cancers –. Immunotherapeutic Strategies for Brain Tumors. Cho CH, Yu J, From Inflammation to Cancer: Brain Tumors - Current and Emerging Advances in Diagnosis & Therapy for Therapeutic Strategies, 331-354, InTech, 2011. Gastrointestinal & Hepatological Diseases. R002. Isomura, H.: DNA polymerase World Scientific Publishing, in press. processivity factor of human cytomegalovirus R011. Osada, H., Takahashi, T.: let-7 and may be a key molecule for molecular coupling of miR-17-92: small-sized major players in lung viral DNA replication to transcription - DNA cancer development. Cancer Sci, 102: 9-17, 2011. Replication, Book 2, 161-178, InTech, 2011. (PMID: 20735434) R003. Kondo, Y., Issa, J.P.: DNA methylation R012. Oze, I., Matsuo, K., Wakai, K., Nagata, profiling in cancer. Expert Rev Mol Med, 12: e23, C., Mizoue, T., Tanaka, K., Tsuji, I., Sasazuki, 2010. (PMID: 20663272) S., Inoue, M., Tsugane, S.; for the Research R004. Kondo, Y.: Chapter 12 - Epigenetic Group for the Development and Evaluation of Changes in Hepatic Tissues: Evidence of Field Cancer Prevention Strategies in Japan.: Alcohol Cancerization. Dakubo GD. Field Cancerization: Drinking and Esophageal Cancer Risk: An Basic Science and Clinical Applications. Nova Evaluation Based on a Systematic Review of Science Publishers. 225-239. 2010. Epidemiologic Evidence Among the Japanese Population. Jpn J Clin Oncol, 41: 677-692, 2011. R005. Matsuo, K., Ito, H., Wakai, K., Nagata, (PMID: 21430021) C., Mizoue, T., Tanaka, K., Tsuji, I., Tamakoshi, A., Sasazuki, S., Inoue, M., Tsugane, S.; for the R013. Sato, Y., Tsurumi, T.: Noise Research Group for the Development and Cancellation: Viral fine tuning of the cellular Evaluation of Cancer Prevention Strategies in environment for its own genome replication. Japan.: Cigar smoking and pancreas cancer risk: PLoS Pathog, 6:e1001158, 2010. (PMID: an evaluation based on a systematic review of 21187893) epidemiologic evidence in thette e Japanese R014. Sekido, Y.: Genomic abnormalities and Population. Jpn J Clin Oncol, 41: 1292-1302, signal transduction dysregulation in malignant 2011. (PMID: 21971423) mesothelioma cells. Cancer Sci, 101: 1-6, 2010. R006. Matsuo, K., Oze, I., Tanaka, Hid.: (PMID: 19793348) Response to Yokoyama et al.: Past and current R015. Sekido, Y.: Inactivation of Merlin in tendency for facial flushing after a small dose of malignant mesothelioma cells and the Hippo alcohol is a marker for increased risk of upper signaling cascade dysregulation. Pathol Int, 61: aerodigestive tract cancer in Japanese drinkers. 331-344, 2011. (PMID: 21615608) Cancer Sci, 101: 2499-2500, 2010. (PMID: 20518787) R016. Seto, M., Honma, K., Nakagawa, M.: Diversity of genome profiles in malignant R007. Nakanishi, H., Ito, S., Matsui, M., lymphoma. Cancer Sci, 101: 573-578, 2010. Murakami, H., Kodera, Y.: In Vivo Cellular (PMID: 20070305) Imaging Using Fluorescent Protein. Methods and Protocols, Hoffman RM (Ed.), Non invasive and R017. Seto, M.: Genomic profiles in B cell real-time fluorescence imaging of peritoneal lymphoma. (Progress in Hematology) Int J metastasis in nude mice. Human Press, Totowa, Hematol, 92: 238-245, 2010. (PMID: 20799004) NJ., in press. R018. Tanaka, M., Katayama, F., Kato, H., R008. Natsume, A., Kondo, Y., Ito, M., Tanaka, Hid., Wang, J., Qiao, Y.L., Inoue, M.: Motomura, K., Wakabayashi, T., Yoshida, J.: Hepatitis B and C virus infection and Epigenetic aberrations and therapeutic hepatocellular carcinoma in china: a review of implications in gliomas. Cancer Sci, 101: epidemiology and control measures. J Epidemiol, 1331-1336, 2010. (PMID: 20384628) 21: 401-416, 2011. (PMID: 22041528) R019. Long, N., Moore, M.A., Chen, W., Gao, R009. Nishio, No., Kojima, S.: Recent progress C.M., Lai, M.S., Mizoue, T., Oyunchimeg, D., in dyskeratosis congenita. Int J Hematol, 92: Park, S., Shin, H.R., Tajima, K., Yoo, K., Sobue,

72 T.: Cancer epidemiology and control in north-East Asia – past, present and future. Asian Pac J Cancer Prev, 11(Suppl 2): 107-148, 2010. (PMID: 20553072) R020. Sonoda, S., Li, H.C., Tajima, K.: Ethnoepidemiology of HTLV-1 related diseases: Ethnic determinants of HTLV-1 susceptibility and its worldwide dispersal. Cancer Sci, 102: 295-301, 2011. (PMID: 21205073) R021. Tajima, K.: General cancer control program in Asia-Pacific countries: Japan as a example. In Cancer Report 2010, Asian Pacific Organization for Cancer Prevention, Tuncer AM, ed, pp147-153, New Hope in Health Foundation, 2010. R022. Tajima, K.: Recent advances of Cancer in Japan. In Recent Advances of Cancer in Asian Countries, The Asian Clinical Oncology Society, ed, pp26-34, Cancer and Chemotherapy Publishers Co, Japan, 2010. R023. Zhou, Z.Y., Matsuo, K., Wang, W.C., Yang, H., Tajima, K., Cao, J.: Dietary risks: Folate, alcohol and gene polymorphisms. In Colorectal Cancer from Prevention to Patient Care, Ettarh R, ed., pp.491-508, INTECH ORG, 2012.

73 H.A., Delamarre, L., Decker, S.A., Ohlfest, J.R., Abstracts for international conferences Okada, H.: COX-2 blockade suppresses gliomagenesis by promoting anti-glioma immunosurveillance. The 101th Annual Meeting A001. Akatsuka, Y.: Characterization and of the American Association of Cancer Research, clinical application of minor histocompatibility Washington, D.C., 2010. antigens. The 15th Annual Winter Meeting of the Korean Society of Blood and Marrow A008. Fujita, M., Scheurer, M.E., Decker, Transplantation, Muju, 2010. S.A., McDonald, H.A., Kohanbash, G., Kato, H., Bondy, M.L., Ohlfest, J.R., Okada, H.: A pivotal A002. Fujii, M., Toyoda, T., Nakanishi, H., role of yype-1 interferon signaling in Yatabe, Y., Sato, A., Matsudaira, Y., Ito, H., immunosurveillance against gliomas - mouse Murakami, H., Kondo, Y., Hida, T., Tsujimura, studies leading to discovery of novel prognostic T., Osada, H., Sekido, Y.:TGF-β synergizes with markers in humans. The 13th Annual Meeting of defects in the Hippo pathway to stimulate human the Translational Research Cancer Centers malignant mesothelioma growth by inducing Consortium, Columbus, 2010. CTGF expression. FASEB SUMMER RESEARCH CONFERENCES: The TGF-β A009. Goto, H., Li, P., Kiyono, T., Matsuyama, Superfamily; Signaling in Development and M., Kasahara, K., Murakami, Y., Yatabe, Y., Disease, Italia, 2011. Inagaki, M.: Chk1 Phosphorylation by p90 Ribosomal S6 Kinase (p90 RSK). The 51th A003. Fujita, M., Decker, S.A., McDonald, Annual Meeting of the American Society for Cell H.A., Kohanbash, G., Ohlfest, J.R., Okada, H.: Biology, Denver, 2011. COX-2 blockade suppresses gliomagenesis by promoting anti-glioma immunosurveillance. The A010. Hirosawa, T.: Is there survival benefits 13th Annual Meeting of the Translational performing retroperitoneal systemic Research Cancer Centers Consortium, Columbus, lymphadenectomy in advanced epithelial ovarian 2010. cancer? 13th Biennial Meeting of the International Gynecologic Cancer Society, A004. Fujita, M., Kohanbash, G., Prague, 2010. Fellows-Mayle, W., Hamilton, R.L., Decker, S.A., Ohlfest J.R., Okada, H.: COX-2 blockade A011. Horio, M., Sato, M., Takeyama, Y., suppresses gliomagenesis by inhibiting Hase, T., Yoshida, K., Sekido, Y., Kondo, M., CCL2-mediated accumulation of Hasegawa, Y.: Knockdown of ZEB1, a master myeloid-derived suppressor cells in glioma epithelial mesenchymal transition (EMT) microenvironment. The 25th Annual Scientific inducing gene, suppresses growth of pleural Meeting of the International Society for mesothelioma cell lines. 10th International Biological Therapy of Cancer, Washington, D.C., Conference of International Mesothelioma 2010. Interest Group, Kyoto, 2010. A005. Fujita, M., Kohanbash, G., A012. Hosono, S., Matsuo, K., Ito, H., Hirose, Fellows-Mayle, W., Hamilton, R.L., Decker, K., Watanabe, M., Nakanishi, T., Tajima, K., S.A., Ohlfest, J.R., Okada, H.: COX-2 blockade Tanaka, Hid.: Polymorphisms in DNA repair suppresses gliomagenesis by inhibiting genes are associated with endometrial cancer risk CCL2-mediated accumulation of among Japanese women. 102nd Annual Meeting myeloid-derived suppressor cells in glioma of the American Association for Cancer Research, microenvironment. The 15th Annual Meeting of Orlando, 2011. the Society for Neuro-Oncology, Montreal, 2010. A013. Inagaki, M.: Novel regulatory A006. Fujita, M., Kohanbash, G., mechanism of Plk1 at mitosis. MEXT Priority Fellows-Mayle, W., Hamilton, R.L., Decker, Research Project International Symposium ”Cell S.A., Ohlfest, J.R., Okada, H.: COX-2 blockade Division”, Hakone, 2011. suppresses gliomagenesis by inhibiting A014. Inoko, A., Ibi, M., Izawa, I.: Trichoplein, myeloid-derived suppressor cells. The 102nd the centriolar Aurora-A partner, prevents primary Annual Meeting of the American Association of cilia. MEXT Priority Research Project “Cell Cancer Research, Orlando, 2011. Proliferation Control” International Symposium A007. Fujita, M., Kohanbash, G., McDonald, Cell Cycle and Cell Differentiation From A to Z, Nagoya, 2010.

74 A015. Ishiguro, F., Murakami, H., Mizuno, T., Microbiological Societies (IUMS) 2011, XV Fujii, M., Kondo, Y., Usami, N., Yokoi, K., Internatinal Congress of Virology, Sapporo, Osada, H., Sekido, Y.: Activated leukocyte cell 2011. adhesion molecule is involved in motility and A023. Kanda, T., Tsurumi, T.: The impact of invasion of malignant pleural mesothelioma. 10th viral repetitive sequences on the biological International Conference of International properties of Epstein-Barr virus recombinants. Mesothelioma Interest Group, Kyoto, 2010. 14th Biennial Conference of the International A016. Isomura, H., Stinski, M.F., Murata, T., Association for Research on Epstein-Barr Virus Kanda, T., Tsurumi, T.: Human cytomegalovirus and Associated Diseases, Birmingham, 2010. late transactivators recruited to the replication A024. Karube, K., Nakagawa, M., Tsuzuki, S., compartments, 35th Annual International Ko, Y.H., Nakamura, S., Seto, M.: Identification Herpesvirus Workshop, Salt Lake City, 2010. of FOXO3 and PRDM1 as tumor suppressor A017. Isomura, H., Tsurumi, T.: Human gene candidates in NK cell neoplasms by the cytomegalovirus genes required for late viral combination of genomic and functional analyses. gene expression and assembly into pre- 11th International Conference on Malignant replication complexes before viral DNA Lymphoma, Switzerland, 2011. replication. 13th CMV / BetaHerpesvirus A025. Karube, K., Seto, M.: Genomic and Workshop, Nuremberg, 2011. functional analyses of NK-cell neoplasms. 4th T A018. Isomura, H., Tsurumi, T.: The human cell lymphoma forum, San Francisco, 2011. cytomegalovirus gene products essential for late A026. Kato, H., Yamamoto, K., Karube, K., viral gene expression assemble into pre- Katayama, M., Tsuzuki, S., Yatabe, Y., replication complexes before viral DNA Takizawa, J., Ohshima, K., Nakamura, S., replication. International Union of Morishima, Y., Seto, M.: Gene expression Microbiological Societies (IUMS) 2011, XV profiling of age-related Epstein-Barr Virus Internatinal Congress of Virology, Sapporo, (EBV)-associated B-cell lymphoproliferative 2011. disorder uncovers alterations in immune and A019. Ito, H., Matsuo, K., Ajiki, W., Sobue, T., inflammatory genes: possible implications for Tanaka, Hid., The Japan Cancer Surveillance pathogenesis. American Society of Hematology Group.: Male breast cancer : a population-mased (ASH) 53rd Annual Meeting, San Diego, 2011. comparison with female breast cancer based on A027. Kawakita, D., Matsuo, K., Hosono, S., data in the monitoring of cancer incidence in Ito, H., Hasegawa, Y., Tanaka, Hid.: Japan project 32nd Annual Meeting of Association between dietary folate intake and International Association of Cancer Registries, clinical outcome in head and neck squamous cell Yokohama, 2010. carcinoma. 3rd World Congress of The A020. Ito, H., Sueta, A., Hosono, S., Iwata, H., International Academy of Oral Oncology, Tajima, K., Tanaka, Hid., Matsuo, K.: A genetic Singapore, 2011. Risk Predictor for Breast Cancer Using A028. Kawakita, D., Sato, F., Hosono, S., Ito, Low-Penetrance Polymorphisms in a Japanese H., Oze, I., Watanabe, M., Hanai, N., Hatooka Population. The 5th Regional Conference of S., Hasegawa, Y., Shinoda, M, Tajima, K., APOCP, Seoul, 2011 Tanaka, Hid., Matsuo, K.: Inverse A021. Kanda, T., Murata, T., Takada, K., Association ,between Yoghurt Intake and Upper Tsurumi, T.: Primary sequence heterogeneity of Aerodigestive Tract Cancer Risk in a Japanese family of repeats (FR) of Epstein-Barr Virus Population. The 5th Regional Conference of (EBV) results in Strain-Specific differences in APOCP, Seoul, 2011. the FR Stability in BAC vectors. 5th Internatinal A029. Kawashima, D., Kanda, T., Tsurumi, T.: Symposium on Nasopharyngeal Carcinoma, Involvement of Hsp90 in Epstein-Barr Virus Penang, 2011. Lytic Replication -Hsp90 facilitates the A022. Kanda, T., Tsurumi, T.: Primary interaction between BALF5 and BMRF1 and Sequence Heterogeneity of Family of Repeats leads to their proper localization-. International (FR) of Epstein-Barr Virus (EBV) results in Union of Microbiological Societies (IUMS) 2011, Strain-Specific Differences in the FR Stability in XV Internatinal Congress of Virology, Sapporo, BAC vectors. International Union of 2011.

75 A030. Kitawaki, T., Kadowaki, N., Fukunaga, 2010. K., Kasai, Y., Maekawa, T., Ohmori, K., A038. Kondo, Y.: Clinical implications of DNA Maekawa, R., Kondo, T., Nieda, M., Yokokawa, methylation in gastrointestinal malignancies. A3 K., Ito, T., Shimizu, A., Tada, H., Kuzushima, Symposium 2010, Jeju, 2010. K., Ishikawa, T., Uchiyama, T.: Phase I/IIa clinical trials of dendritic cell-based A039. Kondo, Y.: Epigenetic networks regulate immunotherapy for acute myeloid leukemia in plasticty of cancer stem cell in glioblastoma. 13th elderly patients using autologous apoptotic Japanese-German Cancer Workshop, Hiroshima, leukemic cells or a heteroclitic WT1 peptide as 2011. antigens. DC2010, Swiss, 2010. A040. Kondo, Y.: Epigenetic plasticity A031. Kitawaki, T., Kadowaki, N., Fukunaga, regulated by PRC-mediated H3K27me3 as a K., Utsumi, M., Kasai, Y., Maekawa, T., Ohmori, novel target for human glioblastoma treatment. K., Maekawa, R., Kondo, T., Nieda, M., JSPS Sweden -Japan Joint Colloquium, Yokokawa, K., Ito, T., Shimizu, A., Tada, H., Stockholm, 2010. Kuzushima, K., Ishikawa, T., Uchiyama, T.: A041. Kondo, Y.: Epigenetic plasticity Phase I/IIa clinical trials of dendritic cell-based regulated by polycomb repressive complex immunotherapy for acute myeloid leukemia in contributes to tissue heterogeneity in human elderly patients. 14th International Congress of glioblastoma. The 41st International Symposium Immunology, Kobe, 2010. of The Princess Takamatsu Cancer Research A032. Kohanbash G, Mintz AH, McKaveney K, Fund, Tokyo, 2010. Fujita M, Okada H.: Roles of interleuikin-4 A042. Kondo, Y.: Epigenetic switch regulates receptor α-chain on glioma-infiltrating pasticty of cancer stem cell in human monocytes. Science 2010, Pittsburgh, 2010. glioblastoma. 6th Asian Epigenomics Meeting, A033. Kohanbash, G., McKaveney, K., Mintz, Kumamoto, 2011. A.H., Scheurer, M.E., Bondy, M.L., Ohlfest JR, . A043. Kondo, Y.: Integrated analysis of genetic Okada H, Fujita M : Roles of interleuikin-4 and epigenetic alterations reveals CpG island receptor α-chain on glioma-infiltrating methylator phenotype associated with clinical monocytes. The 15th Annual Meeting of the outcome of lung adenocarcinoma. Sapporo Society for Neuro-Oncology, Montreal, 2010. Cancer Epigenetics Seminar of the A3 Foresight A034. Kohanbash, G., Mintz, A.H., Program 2011, Sapporo, 2011. McKaveney, K., McDonald, H.A., Ohlfest, J.R., A044. Kondo, Y.: Plastic and stable epigenetic Okada, H., Fujita, M.: Roles of interleuikin-4 regulation in human cancers. Center for Cancer receptor α-chain on glioma-infiltrating Epigenetics, Texas, 2011. monocytes. The 25th Annual Scientific Meeting of the International Society for Biological A045. Li, P.: P90 RSK arranges Chk1 in the Therapy of Cancer, Washington, DC, 2010. nucleus for monitoring of genomic integrity during cell proliferation. Global COE The 3rd A035. Kohanbash, G., Sasaki, K., Hoji, A., International Symposium, Nagoya, 2011 Fujita, M., Okada, H.: miR-17-92 expression in differentiated T cells - implications for cancer A046. Liu, T., Suzuki, M., Nobori, Y.N., Guo, immunotherapy. The 101th Annual Meeting of Y., Hirosawa, N., Ichihara, Y., Sakamoto, Y., the American Association of Cancer Research, Ishihara, O., Kikuchi, H., Senju, S., Uemura,Y.: Washington, D.C., 2010. Distinct subsets of human invariant NKT cells differentially regulate T helper responses via A036. Kohanbash, G., Sasaki, K., Hoji, A., dendritic cells, 14th International Congress of Ueda, R., Fujita, M., Okada, H.:MiR-17-92 Immunology, Kobe, 2010. expression in differentiated T cells – implications for cancer immunotherapy. The 13th Annual A047. Matsuo, K., Kanda, J., Inoue, M., Meeting of the Translational Research Cancer Iwasaki, M., Sawada, N., Shimazu, T., Yamaji, Centers Consortium, Columbus, 2010. T., Sasazuki, S., Tsugane S.: Association of alcohol intake with the risk of malignant A037. Kondo, E.: Development of novel cancer lymphoma and plasma cell myeloma in cell-selective cell-penetrating peptides for the Japanese : a population-based cohort study. 32nd advanced peptide-based drug delivery system. Annual Meeting of International Association of 22nd EORTC-NCI-AACR Symposium, Berlin,

76 Cancer Registries, Yokohama, 2010. cellular factors that enhance reactivation from Epstein-Barr virus latency. 14th Biennial A048. Matsuo, K., Mizoue, T., Tanaka, K., Conference of the International Association for Tsuji, I., Sugawara, Y., Sasazuki, S., Nagata, C., Research on Epstein-Barr Virus and Associated Tamakoshi, A., Wakai, K., Inoue, M., Tsugane, Diseases, Birmingham, 2010. S.: Association between Body-Mass-Index (BMI) and the Colorectal Cancer Risk in Japan: A056. Nakanishi, H., Ito, S., Tsuchida, D., Pooled-Analysis of Population-Based Cohort Kodera, Y.: Molecular aspect of peritoneal Studies in Japan. The 5th Regional Conference of metastasis; A new diagnostic and therapeutic APOCP, Seoul, 2011. strategy in gastric cancer. 9th International Gastric Cancer Congress, Seoul, 2011. A049. Matsuyama, M., Kasahara, K., Goto, H.: Nuclear Chk1 prevents premature activation A057. Nakanishi, H., Oshima, Y., Tsuchida, of cyclinB1-Cdk1. MEXT Priority Research D., Kondo, E.: Molecular targeting and imaging Project “Cell Proliferation Control” International of gastric cancer metastasis. BIT's 3rd Symposium Cell Cycle and Cell Differentiation International Congress of Antibodies-2011 From A to Z, Nagoya, 2010. (ICA-2011), Beijing, 2010. A050. Mizuno, T., Murakami, H., Fujii, M., A058. Nobori, Y.N., Guo, H., Liu, T., Ishiguro, F., Kondo, Y., Akatsuka, S., Toyokuni, Hirosawa, N., Uemura, Y., Jiao, S., Wang, D., S., Ueda, Y., Yokoi, K., Osada, H., Sekido, Y.: Lin, Z., Suzuki, M., Ichihara, Y., Ishihara, O., YAP transcription coactivator induces malignant Senju, S., Kikuchi, H., Sakamoto, Y., Sonoda, mesothelioma cell proliferation by up-regulating Y.: Bisphenol A in combination with TNFa cell cycle progression. The 14th World selectively induces Th2 cell-promoting dendritic Conference on Lung Cancer, Amsterdam, 2011. cells in vitro with an estrogen-like activity. 14th International Congress of Immunology, Kobe, A051. Mizuno, T., Murakami, H., Fujii, M., 2010. Taniguchi, T., Ishiguro, F., Kondo, Y., Akatsuka, S., Toyokuni, S., Ueda, Y., Yokoi, K., A059. Noda, C., Murata, T., Kanda, T., Osada, H., Sekido, Y.: YAP induces malignant Tsurumi, T.: Identification and Characterization mesothelioma cell proliferation via induction of of a Novel Transcriptional Activator for EBV CCND1 expression. 10th International Oncogene LMP1. International Union of Conference of International Mesothelioma Microbiological Societies (IUMS) 2011, XV Interest Group, Kyoto, 2010. Internatinal Congress of Virology, Sapporo, 2011. A052. Murakami, H., Kawata, S., Taniguchi, T., Kawaguchi, K., Mizuno, T., Ishiguro, F., A060. Noguchi, K., Yasuda, A., Katayama, K., Fujii,M.,Kondo,Y.,Osada,H.,Sekido,Y.: Mitsuhashi, J., Kanda, T., Minoshima, M., Expression and functional analysis of Hairy Bando, T., Sugiyama, H., Sugimoto, Y.: Enhancer of Split 1 (HES1) in human malignant EBV-induced Immortalization Is Inhibited by mesothelioma cell lines. 10th International Polyamide Targeting EBNA1-oriP Binding. Conference of International Mesothelioma International Union of Microbiological Societies Interest Group, Kyoto, 2010. (IUMS) 2011, XV Internatinal Congress of Virology, Sapporo, 2011. A053. Murata, T., Noda, C., Kanda, T., Tsurumi, T.: Identification and characterization A061. Ohka, F., Natsume, A., Motomura, K., of a novel transcriptional activator for EBV Kishida, Y., Kondo, Y., Momota, H., Iwami, K., oncogene LMP1 in epithelial cells. 5th Kinjo, S., Ito, M., Fujii, M., Wakabayashi, T.: Internatinal Symposium on Nasopharyngeal The global DNA methylation surrogate LINE-1 Carcinoma, Penang, 2011. methylation is correlated with MGMT Promoter methylation and is a better prognostic factor for A054. Murata, T., Tsurumi, T.: Involvement of glioma. Sapporo Cancer Epigenetics Seminar of Jun Dimerization Protein 2 (JDP2) in the the A3 Foresight Program 2011, Sapporo, 2011. Maintenance of Epstein-Barr virus Latency. International Union of Microbiological Societies A062. Okada, H., Sasaki, K., Kohanbash, G., (IUMS) 2011, XV Internatinal Congress of Hoji, A., Fujita, M., Ueda, R., McDonald, H.A., Virology, Sapporo, 2011. Reinhart, T.A., Martinson, J., Lotze, M.T., Wang, E., Marincola, F.M.: Regulation of A055. Murata, T., Tsurumi, T.: Screening of miR-17-92 expression in differentiated T cells -

77 implications for cancer immunotherapy. The 18th mesothelioma. 10th International Conference of International Conference on Brain Tumor International Mesothelioma Interest Group, Research and Therapy, Travemünde, 2010. Kyoto, 2010. A063. Okamoto, S., Ikeda, H., Fujiwara, H., A070. Sekido, Y.: Dysregulation of signal Nukaya, I., Yoshioka, H., Kuzushima, K., transduction cascades in malignant mesothelioma Yasukawa, M., Shiku, H., Mineno, J.: Highly cells induces connective tissue growth factor efficient and safe vectors for TCR gene therapy: (CTGF) expression. 9th China-Japan Joint silencing of endogenous tcr improved expression Conference of Cancer, Shanghai, China, 2011. and anti-tumor reactivity of transduced A071. Sekido, Y.: LATS2 is a tumor suppressor tumor-specific tcrs in human lymphocytes. gene of malignant mesothelioma. 2nd Workshop American Society of Gene and Cell Therapy, on the Hippo Tumor Suppressor Pathway, Italia, Seattle, 2011. 2010. A064. Okamoto, Y., Shinjo, K., Tanaka, Y., A072. Shinjo, K., Kondo, Y., Okamoto, Y., Fujii, M., Murakami, H., Osada, H., Kataoka, Yokoyama, T., Usami, N., Fujii, M., Murakami, H., Joh, T., Sekido, Y., Kondo, Y.: Analysis of H., Osada, H., Sekido, Y.: Integrated analysis of epigenetic alterations in mice with humanized genetic and epigenetic alterations reveals CpG liver after hepatitis virus infection. 102nd AACR island methylator phenotype associated with annual meeting, Orlando, 2011. clinical outcome of lung adenocarcinoma. 102nd A065. Osada, H., Nishikawa, E., Arima, C., AACR annual meeting, Orlando, 2011. Okazaki, Y., Tomida, S., Tatematsu, Y., Taguchi, A073. Shinjo, K., Okamoto, Y., Fujii, M., A., Shimada, Y., Yanagisawa, K., Toyokuni, S., Murakami, H., Osada, H., Sekido, Y., Kondo, Sekido, Y., Takahashi, T.: Roles of Y.: Significance of CpG island methylator ASH1-miR-375 pathway in development of lung phenotype in human lung cancers. JSPS Sweden cancers with neuroendocrine features. 102nd -Japan Joint Colloquium, Stockholm, 2010. AACR Annual Meeting 2011, Orlando, 2011. A074. Sugimoto, A., Nishiyama, Y., Tsurumi, A066. Oze, I., Matsuo, K., Hosono, S., Ito, H., T.: Anatomy of Epsterin-Barr virus genome Watanabe, M., Tajima, K., Tanaka, Hid.:The manufacturing plant. International Union of association between coffee and green tea intake Microbiological Societies (IUMS) 2011, XV and upper aerodigestive tract cancer in Japanese Internatinal Congress of Virology, Sapporo, population. 102nd Annual Meeting of the 2011. American Association for Cancer Research, Orlando, 2011. A075. Sugimoto, A., Nishiyama, Y., Tsurumi, T.: Spatiotemporally deffernt DNA repair A067. Oze, I., Matsuo, K., Hosono, S., Ito, H., systems participating during Epstein-Barr virus Watanabe, M., Tajima, K., Tanaka, Hid.: Facial genome maturation. Structural Biology & DNA Flushing and ALDH2 Glu504Lys Polymorphism Repair, Amsterdam, 2011. after Alcohol Consumption for the Risk of Upper Aerodigestive Tract Cancer Risk in a Japanese A076. Suzuki, M., Uemura, Y., Liu, T., Nobori, Population. The 5th Regional Conference of Y.N., Hirosawa, N., Uchino, S., Ichihara, Y., APOCP, Seoul, 2011. Kikuchi, H., Sakamoto, Y., Senju, S., Ishihara, O.: Role of human non-invariant NKT A068. Oze, I., Matsuo, K., Hosono, S., lymphocytes in the maintenance of type 2 T Watanabe, M., Ito, H., Tajima, K., Tanaka Hid.: helper environment during pregnancy. 14th Positive association between green tea International Congress of Immunology, Kobe, consumption and risk for upper aerodigestive 2010. tract cancer in Japanese population. 32nd Annual Meeting of International Association of Cancer A077. Takahashi, Y., Doisaki, S., Nishio, No., Registries, Yokohama, 2010. Muramatsu, H., Shimada, A., Hama, A., Kojima, S.: The telomere length in peripheral blood A069. Sekido, Y., Mizuno, T., Taniguchi, T., lymphocytes is a useful predictor of the response Fujii, M., Ishiguro, F., Fukui, T., Akatsuka, S., to immunosuppressive therapy in patients with Horio, Y., Hida, T., Kondo, Y., Toyokuni, S., acquired aplastic anemia. 52th annual meeting of Osada, H., Murakami, H.: Inactivation of american society of hematology, Orlando, 2010. LATS2 indicates frequent dysregulation of the Merlin-Hippo signaling pathway in malignant A078. Tanaka, Hid., Hosono, S.: Estimation of

78 incidence rate of hepatitis b virus hopizontal A086. Yamamura, T., Bleakley, M., Hikita, J., infection in the Japanese population in 2005. IEA Matsubara, A., Hamajima, T., Nannya, Y., World Congress of Epidemiology, Edinburgh, Takahashi, T., Emi, N., Morishima, Y., Kodera, 2011. Y., Kuzushima, K., Riddell, S.R., Ogawa, S., Akatsuka, Y.: Development of an online tool to A079. Tanaka, Hid., Saka, H., Amagai, K., scan single nucleotide polymorphisms for Tanaka, M., Katanoda, K., Taniguchi, C.: identification of novel minor histocompatibility Proceeding of 'Tabacco Contriol Planning' for 32 antigens. 17th BMT Tandem Meetings, Honolulu, cancer hospitals designated by Japanese Ministry 2010. of Health, Labor and Welfare. Asia Pacific Conference on Tobacco or Health 2010, Sydney, A087. Zhang, R., Ito, S., Nishio, Na., Cheng, 2010. Z., Suzuki, H., Isobe, K.,: Increase of myeloid derived suppressor cells (MDSC) in spleen of A080. Taniguchi, C., Hibino, F., Minami, M., dextran sulphate sodium-treated C57BL/6 mice. Hyoudou, C., Maruguchi, M., Tokunaga, N., 14th International Congress of Immunology, Saka, H., Tanaka, Hid.: Tobacco intervention Kobe, 2010. attitudes and practices among Japanese nurses for cancer patientsAsia Pacific Conference on A088. Tajima, K.: Global cancer control Tobacco or Health 2010, Sydney, 2010. direction with special reference to Asia-Pacific area. 5th Regional Conference of APOCP, Seoul, A081. Truong, T., Lathrop, M., Brennan, P., 2011. on behalf of the INHANCE consortium.: A genome-wide association study of upper A089. Tajima, K., Trapido, E.: Translating aerodigestive tract cancers identifies 4q21, 4q23, epidemiological evidence into policy & practice. and 12q24 as susceptibility loci. American 4th International Cancer Control Congress, Seoul, Association for Cancer Research 101st 2011. ANNUAL MEETING 2010, Washington, 2010. A090. Tajima, K.: Ethnoepidemiology of Asian A082. Tsurumi, T., Nakayama, S., Murata, T.: cancer-epidemic pattern and its prevention Tail to tail contact for tetrameric ring formation strategy. 21st Asia Pacific Cancer Conference, of EBV processivity factor is crucial for viral Kuala Lumpur, 2011. replication. The EMBO meeting 2010, Barcelona, A091. Tajima, K.: Ethnoepidemiologic study on 2010. main cancers in Korea, Japan and China A083. Uemura, Y., Liu, T., Nobori, Y.N., (KOJACH Cooperative Study) (Symposium 9: Suzuki, M., Hirosawa, N., Ichihara, Y., Research Collaboration). The 6th General Ishihara, O., Kikuchi, H., Sakamoto, Y., Sonoda, Assembly of the Asian Pacific Organization for Y., Senju, S.: Cytokine-dependent modification Cancer Prevention, Sarawak, 2012. of IL-12p70 and IL-23 balance in dendritic cells A092. Tajima, K.: International comparative by ligand activation of Va24 invariant natural epidemiologic study on main cancers in Korean, killer T cells. 14th International Congress of Japan and China (KOJACH Cooperative Study). Immunology, Kobe, 2010. (SL). The 2nd Japan-China Symposium on Cancer A084. Umino, A., Nakagawa, Masao., Research, Chiba, 2012. Utsunomiya, A., Tsukasaki, K., Katayama, N., Seto, M.: Array comparative genomic hybridization revealed polyclonality in acute type adult T-cell leukemia/lymphoma and PTCL NOS. 52nd ASH Annual Meeting and Exposition, Orlando, 2010. A085. Yamamoto, K., Tsuzuki, S., Naoe, T., Seto, M.: Deregulated activity of AML1/RUNX1 cooperates with BCR-ABL to immortalize hematopoietic progenitor cells and induces blast crisis-like disease of chronic myelogenous leukemia in mice. American Society of Hematology (ASH) 53rd Annual Meeting, San Diego, 2011.

79 Record of Seminars ______Invited Speakers 2010 Feb. 19 Shimono, Y. (Stem Cell Institute, Stanford University): MicroRNA Profiling Uncovers 'Stemness" of Human Breast Cancer Stem Cells.

May. 08 Suzuki, H. (First Department of Internal Medicine, Sapporo Medical University): Epigenetic silencing mechanisms of microRNA in gastric cancers.

May. 12 Aoki, M. (Department of Pharmacology, Graduate School of Medicine, Kyoto University): Studies on signaling pathways involved in carcinogenesis.

May 14 Maeda, T. (Department of Hematopoietic Stem Cell & Leukemia Research, Beckman Research Institute of the City of Hope): Role of targeting proteins related to cellular transformation.

May 18 Ohtani, N. (Division of Cancer Biology, The Cancer Institute, Japanese Foundation for Cancer Research): Mechanism of cellular senescence and tumor suppression.

Jun 24 Matsuo, K. (Division of Epidemiology and Prevention, Aichi Cancer Center): Genome-wide association study for drinking behavior.

Jul. 27 Keyomarsi, K. (Department of Experimental Radiation Oncology, University of Texas, MD Anderson Cancer Center): Personalized therapy for breast cancer by targeting the cell cycle -Examining the deregulation of cyclin E in breast cancer metastasis and generation of breast cancer stem cells-.

Nov. 29 Kishi, H. (Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama): A rapid and efficient single cell manipulation method for screening antigen-specific antibody-secreting cells and T-cell receptors.

Dec. 02 Miyahara, Y. (Department of Immuno-Gene Therapy, Mie University Graduate School of Medicine): Development and immunotolerance of Foxp3+ regulatory T cells.

Dec. 27 Sawyers, C.L. (Memorial Sloan-Kettering Cancer Center): Overcoming Resistance to Targeted Cancer Therapies.

2011 Jan 17 Fujita, M. (Brain Tumor Program, University of Pittsburgh Cancer Institute): Anti-glioma immunosurveillance and development of effective immunotherapeutic strategies using glioma model mice induced by the Sleeping Beauty (SB) transposon system.

Jan 21 Miyoshi, H. (Department of Pathology and Immunology, Washington University School of Medicine): Colonic crypt morphogenesis after injury by the mesenchymal niche.

Nov. 16 Oki, M. (Department of Applied Chemistry & Biotechnology, Graduate school of Engineering, University of Fukui): epigenetic mechanism controlling boundaries of transcriptionally silent heterochromatin in Saccharomyces cerevisiae.

80 Nov. 22 Pan, D. (Department of Molecular Biology & Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland Investigator, Howard Hughes Medical Institute): Control of organ size and tumorigenesis by the Hippo signaling pathway.

Dec. 10 Inoue, M. (Department of Biochemistry, Osaka Medical Center for Cancer and Cardiovascular Diseases): Novel culture methods of primary cancer cells: Development and Applications.

Institute Speakers 2010 Feb. 18 Ibi, M. (Biochemistry): A keratin-intermediate filament scaffold protein trichoplein controls microtubule anchoring at the centrosome.

May. 13 Kondo, Y. (Molecular Oncology): Epigenetics in cancer ‐Basic research and clinical applications‐.

Oct. 29 Tsuzuki, S. (Molecular Medicine): Expression of hematopoietic stem and progenitor cells by a short isoform of AML1/RUNX1.

Nov. 18 Hirosawa, T. (Immunology): Development of immunotherapy against ovarian cancer using cytotoxic T lymphocytes.

Dec. 22 Murata, T. (Virology): EBV latency, reactivation and cell growth.

2011 Jan. 27 Sakuma, K. (Molecular Pathology): Mechanisms of selectin-ligand glycan expression in colon cancer cells undergoing EMT.

Feb. 17 Matsuyama, M. (Biochemistry): Nuclear Chk1 prevents premature mitotic entry.

Mar. 17 Nakanishi, H. (Oncological Pathology): Basic research on the molecular targeting therapy and molecular imaging of the gastrointestinal malignancies and its clinical applications.

May. 19 Nakagawa, M. (Molecular Medicine): Synergistic functional effect of oncogenes in B cell lymphoma.

June. 27 Saito, K. (Oncological Pathology): Effector Molecule of Cell Death in Gefitinib Response NSCLC and Functional Analysis of OGFOD1 as a Novel Molecular Target in Tumor.

June. 28 Nishio, No. (Immunology): Ex vivo-expanded gammadelta T cells are a promising tool for anti-neuroblastoma immunotherapy.

Sep. 8 Fujii, M. (Molecular Oncology): TGF-β synergizes with defects in the Hippo pathway to stimulate human malignant mesothelioma growth.

Dec. 22 Noda, C. (Virology): Identification and characterization of novel host factors that regulate EBV LMP1.

81 ______Record of Symposia The 15th Aichi Cancer Center International Symposium “New Molecular Target Therapy and Signal Transduction”

Organizing Committee: Yoshitaka Sekido (Chairperson), Tetsuya Mitsudomi, Toyoaki Hida, Kei Muro, Shinobu Tsuzuki, Hidemasa Goto, Makiko Fujii, Kazuo Tajima

March 6, 2010, International Conference Hall, Aichi Cancer Center.

Program of symposium

Opening Remarks: Kazuo Tajima (Aichi Cancer Center)

Session 1. Discoveries of New Target Molecules for Cancer Therapy Chairperson: Mutsuhiro Takekawa (Nagoya University) Hidemasa Goto (Aichi Cancer Center Research Institute) Regulation of Chk1 by novel phosphorylations; Perspective of Chk1 as a therapeutic target for cancer Vinay Tergaonkar (Institute for Molecular and Cell Biology, Singapore) New Controls of the NFkB Pathway

Session 2. Recent Advances in Understanding Signal Transduction Cascades Chairperson: Masahide Takahashi (Nagoya University) Hidenori Ichijo (The University of Tokyo) ASK family Signaling in Stress and Disease Kun-Liang Guan (University of California, USA) Regulation of the YAP oncoprotein by the Hippo tumor suppressor Pathway Takao Yamori (Japanese Foundation for Cancer Research) A New PI3 Kinase Inhibitor ZSTK 474 identified by JFCR39 Cancer Cell Lines Panel

Luncheon Seminar Chairperson: Tetsuya Mitsudomi (Aichi Cancer Center Hospital) Pasi A. Janne (Dana- Farber Cancer Institute, USA) Resistant Mechanisms to EGFR Targeted Therapies in Non-Small Cell Lung Cancer

Session 3. EGFR Targeting Therapy in Lung and Colorectal Cancer Chairperson: Yoshinori Hasegawa (Nagoya University) Seiji Yano (Kanazawa University) HGF-induced resistance to reversible and irreversible EGFR-TKIs in lung cancer harboring EGFR mutations Tae Won Kim Asan Medical Center, Korea Biomarkers for Cetuximab in Korean Colorectal Patients Tomoya Yokota (Aichi Cancer Center Hospital) KRAS/BRAF mutations and PTEN/p-AKT expression as predictive markers for Cetuximab in Japanese colorectal cancer patients

Session 4. Invention of New Molecular Target Therapies Chairperson: Shinsuke Iida (Nagoya City University) Hiroyuki Mano (The University of Tokyo) Discovery and clinical application of a novel oncogene, EML4-ALK, in lung cancer Jennifer A. Low (Genentech, South San Francisco, USA) Inhibition of the Hedgehog Signaling Pathway in Cancer Yutaka Kondo (Aichi Cancer Center Research Institute) Targeting Epigenetic Plasticity as a Novel Treatment for Human Neoplasia.

Closing Remarks: Yoshitaka Sekido (Aichi Cancer Center Research Institute)

82 Abstracts

Regulation of Chk1 by novel New Controls of the NFkB Pathway phosphorylations; Perspective of Chk1 Vinay Tergaonkar as a therapeutic target for cancer Institute for Molecular and Cell Biology, Singapore Hidemasa Goto, Masato Enomoto, Kousuke The transcription factor NFκB is critical for Kasahara & Masaki Inagaki several cellular and developmental events in Division of Biochemistry, Aichi Cancer Center metazoans. Given that over 200 physiological Research Institute stimuli activate NFκB, which in turn regulates an Cell cycle is controlled by several families of equally large number of genes, understanding how protein kinases, such as Cyclin-dependent kinases specificity is generated in such a pleiotropic (Cdks) or Checkpoint kinases (Chks). Their pathway is also a major challenge. Using a activities orchestrate the complex events that drive large-scale functional genomics approach, parallel the cell cycle, and are frequently deregulated in interrogation of approximately 20,000 cancer cells. In response to DNA damage, Chk1, sequence-annotated genes was carried out and which is activated by ATR, phosphorylates and several novel modifiers of NFκB activity were inhibits Cdc25 family phosphatases, thus blocking identified. I will describe the characterization of the activation of Cdk1 (Cyclin-dependent kinase 1) some of these modulators and the mechanisms by and preventing premature mitotic entry. Here, we which they regulate NFκB in inflammation and show functional changes of Chk1 by two types of cancer. novel phosphorylations. 1) Ser286 and Ser301 phosphorylation by Cdk1 at ASK family Signaling in Stress and Disease G2/M transition: Chk1 is phosphorylated at Ser286 Hidenori Ichijo and Ser301 by Cdk1 during mitosis. Chk1 Laboratory of Cell Signaling, Graduate School of phosphorylation by Cdk1 regulates Chk1 export Pharmaceutical Sciences, The University of Tokyo from nucleus at G2/M transition. This elimination Actions of pro- and anti-apoptotic factors are of Chk1 activity from nucleus induces the adequate often modulated by phosphorylation and activation of Cdk1 in the nucleus, which regulates dephosphorylation, and protein kinases and protein proper mitotic entry. Our data indicate a novel phosphatases thus contribute to the regulation of positive feedback loop between Chk1 and Cdk1 in cell death decisions made in response to various the nucleus at G2/M transition. stresses. Apoptosis Signal-regulating Kinase 1 2) Ser296 autophosphorylation during DNA (ASK1) is a member of the mitogen-activated damage response: ATR induces Chk1 protein (MAP) kinase kinase kinase family, which phosphorylation at Ser317 and Ser345, which activates both the MKK4/MKK7-JNK and promotes Chk1 catalytic activation at damage sites. MKK3/MKK6-p38 MAP kinase pathways and Ser296 autophosphorylation first occurs in this constitutes a pivotal signaling pathway in various ATR-activated (restricted) area but promotes rapid types of stress responses. ASK1 plays crucial roles dephosphorylation at the ATR sites, which enables in oxidative stress- and endoplasmic reticulum (ER) the release of Chk1 from the restricted area. This stress-induced apoptosis and calcium signaling that shift in Chk1 phosphorylation site(s) not only plays are implicated in the pathophysiology of a broad an important role in the spread of Chk1 signals but range of human diseases. Moreover, ASK1-p38 also changes Chk1-binding subtype of 14-3-3 (from pathway was recently found to play important roles β ζ γ γ or to ). 14-3-3 works as a platform between in the innate immune responses. In this symposium, Cdc25A and Ser296-phosphorylated Chk1, I will review our recent findings on the redox promoting Chk1-induced Cdc25A phosphorylation control and in vivo roles of ASK family kinases in at Ser76, a critical site for its degradation. So, our stress responses. I will also summarize the roles of data precisely demonstrate how Chk1 relays the stress-activated MAP kinase pathways and update signals from ATR to Cdc25A. our new findings on the pathophysiological roles of We also discuss the above phosphorylations ASK family kinases in stress responses and disease. from the standpoint of the screening of new Chk1 inhibitors. Regulation of the YAP oncoprotein by the Hippo tumor suppressor pathway Kun-Liang Guan

83 Department of Pharmacology and Moores Cancer ZSTK474 bound to the ATP-binding pocket of Center, University of California at San Diego, CA, p110, the catalytic subunit of PI3K. ZSTK474 did USA not inhibit protein kinases (> 140) including mTOR The YAP is a transcription co-activator and a and DNA-PK, indicating that ZSTK474 is very candidate human oncogene. YAP is amplified in specific to PI3K. Concerning the specificity to PI3K some human cancers and plays a key role in organ isoforms (alpha, beta, delta and gamma), ZSTK474 size regulation. We showed that YAP is turned out to be a pan-PI3K inhibitor. Furthermore, phosphorylated and inhibited by the Hippo tumor we found that ZSTK474 had antiangiogenic activity. suppressor pathway. Phosphorylation of YAP S127 It inhibited both VEGF secretion from cancer cells by the Lats tumor suppressor kinase results in and angiogenic functions of endothelial cells. 14-3-3 binding and cytoplasmic localization, ZSTK474 showed remarkable antitumor effect in therefore leading to YAP inhibition. various human cancer xenograft models without Phosphorylation of YAP S381/S384 by Lats and serious toxicities. Interestingly ZSTK474 induced CK1δ/ε recruits the SCFβ-TRCP E3 ubiquitin ligase, not apoptosis but strong G1 arrest, which may be which then promotes YAP ubiquitination and the mechanism of its therapeutic efficacy. These degradation. The S127 phosphorylation dependent results indicated that ZSTK474 is a promising translocation and the phosphodegron-mediated candidate of PI3K-targeting drug. It was also degradation coordinately suppress YAP oncogenic proved that JFCR39 is a powerful tool to identify activity. Furthermore, we have established that the molecular target of a new chemical and to the TEAD family transcription factors play a key promote its translational research. role in mediating the biological functions of YAP. Together, our studies establish a mechanism of HGF-induced resistance to reversible and spatial and temporal regulation of YAP by the irreversible EGFR-TKIs in lung cancer Hippo tumor suppressor pathway and their role in harboring EGFR mutations tumorigenesis. Seiji Yano Division of Medical Oncology, Cancer Research A New PI3 Kinase Inhibitor ZSTK474 Institute, Kanazawa University identified by JFCR39 Cancer Cell Lines Non-small cell lung cancer (NSCLC) with Panel epidermal growth factor receptor (EGFR)-activating Takao Yamori mutations (EGFRmu) responds favorably to the Division of Molecular Pharmacology, Cancer EGFR tyrosine kinase inhibitors (EGFR-TKIs), Chemotherapy Center, Japanese Foundation for gefitinib or erlotinib. However, 25–30% of patients Cancer Research with EGFRmu show intrinsic resistance, and the Phosphatidylinositol 3-kinase (PI3K) is a lipid responders invariably acquire resistance to kinase playing a key role in the downstream EGFR-TKIs. Recently, two mechanisms, signaling of receptor tyrosine kinases. Genetic second-site point mutation that substitutes aberration and activation in the PI3K pathway have methionine for threonine at position 790 (T790M) been detected in various human cancers, implying in EGFR and amplification of MET proto-oncogene, that PI3K promotes growth and progression of which contribute to acquired resistance to cancer. Therefore, PI3K is a promising target for EGFR-TKIs have been reported. T790M secondary cancer chemotherapy. Development of PI3K mutation and MET amplification are found in inhibitors has recently become a hot area. We approximately 50% and 20%, respectively, of developed a novel triazine derivative ZSTK474 patients acquiring resistance to EGFR-TKIs. The with strong cancer cell growth inhibition. However, mechanisms of intrinsic resistance and the other its molecular target was unknown. To identify the 30% of cases of acquired resistance are, however, target, we utilized COMPARE algorithm based on still unknown. We recently demonstrated that chemosensitivity measurements from 39 human hepatocyte growth factor (HGF) induced resistance cancer cell lines (JFCR39). We previously indicated to both reversible and irreversible EGFR-TKIs. that JFCR39 with COMPARE could predict the HGF, a specific ligand of MET, induced the unknown molecular targets of novel chemicals. The resistance by restoring Akt phosphorylation predicted most probable target of ZSTK474 was independently of ErbB3. Importantly, combined use PI3K, which was then verified by experiments. of gefitinib plus anti-HGF antibody or the HGF ZSTK474 inhibited the phosphorylation of PI3K antagonist, NK4 successfully overcame the downstream components in cancer cells in vitro and HGF-induced EGFR-TKI resistance both in vitro in vivo. Subsequent analyses revealed that and in vivo. Our findings indicate that

84 HGF-mediated MET activation is a pivotal Tomoya Yokota1, Noriko Shibata2, Yu Sakai2, mechanism of intrinsic and acquired resistance to Takashi Ura1, Daisuke Takahari1, Kohei Shitara1, reversible and irreversible EGFR-TKIs in NSCLC Takeshi Shibata1, Kei Muro1 & Yasushi Yatabe2 with EGFRmu. Therefore, inhibition of HGF-MET 1Clinical Oncology, and 2Pathology and Molecular signaling may be considerable strategy for more Diagnostics, Aichi Cancer Center Hospital successful treatment with EGFR-TKIs. Background: Alterations in Ras/Raf/ERK and PTEN/PI3K/AKT signaling pathways are Biomarkers for Cetuximab in Korean frequently observed in colorectal cancer (CRC). Colorectal Patients This study aims to investigate KRAS/BRAF Tae Won Kim mutation frequency in Japanese CRC patients and Department of Oncology, Asan Medical Center, validate the predictive value of PTEN/p-AKT University of Ulsan College of Medicine, KOREA expression on response to Cetuximab, anti-EGFR The epidermal growth factor receptor (EGFR) monoclonal antibody. has an integral role in cell survival signaling. Patients and methods: Genotyping of KRAS and Specific monoclonal antibodies, such as cetuximab, BRAF mutations was performed on 328 and 307 bind competitively to the extracellular domain of CRC patients, respectively, by Cycleave PCR. As EGFR, inhibiting EGF binding and receptor of November 2009, the expression of PTEN/p-AKT autophosphorylation. Cetuximab has been approved in 25 CRC patients was analyzed by for the treatment of metastatic colorectal cancer immunohistochemistry (IHC). (mCRC) patients. A significant number of patients Results: Although KRAS mutations were present do not benefit from cetuximab treatment. Therefore, in 116 patients (35.4%), only 2.6% (n = 8) of it is necessary to identify novel molecular and patients carried BRAF mutation. The primary biological markers that can be effectively used to lesions of the 5 patients with BRAF mutation were predict patient response. The KRAS gene, one of located on the right side of the colon. The the cetuximab predictor candidates, is mutated in pathological features of CRC patients with BRAF 27-43% of CRC patients. Recent retrospective data mutation included poorly differentiated analyses have indicated that KRAS mutations are adenocarcinoma (4/8) and mucin producing tumor significantly associated with resistance to (2/8). Two patients with BRAF mutation were cetuximab and shorter survival times after treated with cetuximab-combined chemotherapy, cetuximab treatment. but not responded. Low PTEN expression was We have investigated the clinical relevance of associated with increased response to Cetuximab KRAS mutational status in 66 irinotecan-refractory compared with high expression, but not statistically Korean mCRC patients treated with significant (50% vs. 18%, p=0.0999). The response cetuximab-plus-irinotecan-based chemotherapy. A rate in patients with low p-AKT expression was KRAS mutation was detected in 27 (40.9%) tumors, higher than in those with high expression (58% vs. and was associated with lower RR and shorter 22%, p=0.098). However, PTEN/p-AKT expression progression-free survival and overall survival. The level was not significantly associated with KRAS mutation types and frequencies in our series progression-free survival. are similar to those previously reported in Western Conclusion: BRAF mutation frequency in CRC populations. Our results indicate that neither KRAS patients was lower than expected. mutation frequency nor the role of KRAS as a Clinicopathological features of CRC patients with predictive marker is influenced by ethnic BRAF mutations seem to be distinct from those differences. with wild type BRAF. The role of PTEN/p-AKT A number of markers, such as EGFR protein status as a predictive marker for Cetuximab is expression, EGFR gene copy number, and unclear. Further investigation is ongoing. expression of proteins downstream in the EGFR signaling pathway, have been investigated as Discovery and clinical application of a potential predictors of response to cetuximab. novel oncogene, EML4-ALK, in lung Role of FCR polymorphism, BRAF mutation, and cancer EGFR FISH will be discussed. Hiroyuki Mano Department of Medical Genomics, Graduate School KRAS/BRAF mutations and PTEN/p-AKT of Medicine, The University of Tokyo expression as predictive markers for Division of Functional Genomics, Jichi Medical Cetuximab in Japanese colorectal cancer University patients Lung cancer remains the leading cause of cancer

85 deaths worldwide. Conventional chemotherapeutic Basal Cell Nevus (Gorlin) Syndrome. In this regimens improve only marginally the outcome of syndrome, patients with mutations in the Patched individuals with non-small-cell lung cancer receptor develop hundreds or thousands of basal (NSCLC), resulting in a median survival time of <1 cell carcinomas and are at increased risk of year. In order to discover novel transforming genes developing medulloblastoma, a brain cancer of in lung cancer, we have developed a sensitive childhood. Inappropriate hedgehog pathway retroviral expression library system for full-length signaling in associated stromal tissue has also been cDNAs, and applied this technology to an NSCLC seen in epithelial malignancies such as colorectal, specimen that did not harbor EGFR mutations. A ovarian, and pancreatic tumors. Inhibition of focus formation assay with the library led to the hedgehog pathway signaling has long been discovery of a novel fusion gene between the theorized as a potential anticancer target and the echinoderm microtubule associated protein-like 4 natural product, cyclopamine, has been used in the (EML4) and the anaplastic lymphoma kinase laboratory to evaluate hedgehog pathway inhibition. (ALK) genes, both of which are closely mapped to Genentech and other companies have recently the same short arm of chromosome 2. Fusion to developed clinical hedgehog pathway inhibitors. EML4 induces a constitutive dimerization of the GDC-0449 is a hedgehog pathway inhibitor that is ALK kinase domain, and thereby its marked being tested in the clinic, and has recently been activation. reported to have a well-tolerated adverse event Transgenic mice expressing EML4-ALK in lung profile, high sustained plasma drug levels, and epithelial cells generate hundreds of indications of tumor regressions in a high adenocarcinoma nodules soon after birth, proportion of basal cell carcinomas and in a patient demonstrating the pivotal role of this fusion kinase with medulloblastoma. There are ongoing clinical in NSCLC. Further, oral administration of a specific trials in a variety of other tumor types with inhibitor to ALK successfully cleared such nodules GDC-0449 and other clinical hedgehog pathway from the mice. Similar ALK inhibitors to treat inhibitors. The hedgehog pathway is a validated patients are now under development among many oncology clinical target that shows promise for the pharmaceutical companies, and one of such future treatment of cancer. inhibitor has been already under clinical trials for NSCLC with EML4-ALK. Knowing the high Targeting Epigenetic Plasticity as a Novel therapeutic efficacy of such compound, we have Treatment for Human Neoplasia developed an efficient diagnositic scheme for the Yutaka Kondo1, Atsushi Natsume2 & Yoshitaka patients with EML4-ALK, coupling a multiplex Sekido1 RT-PCR with a sensitive immunohistochemical 1 Division of Molecular Oncology, Aichi Cancer detection technology. By utilizing such protocols, Center Research Institute we have constructed a nation–wide diagnostic 2 Department of Neurosurgery, Nagoya University system for EML4-ALK–positive NSCLC in Japan, School of Medicine and have unexpectedly discovered another fusion of Cancers are mostly comprised of heterogeneous ALK, KIF5B-ALK, in a subset of NSCLC. cell populations. A significant degree of These data demonstrate that a subset of lung morphological and lineage heterogeneity may cancer express previously unidentified fusion contribute to tumor expansion, invasion, metastasis kinases that are promising candidates for a and drug resistance. However, the molecular therapeutic target as well as for a diagnostic mechanisms underlying tumor heterogeneity and molecular marker in this intractable disorder. associated plasticity are unknown. Using glioblastoma as a model, we studied epigenetic Inhibition of the Hedgehog Signaling patterns in glioma-initiating cells (GICs) that have Pathway in Cancer the potential to convert into differentiated cell types. Jennifer A. Low We found that biologic conversion of GICs is Genentech BioOncology, South San Francisco, CA, reversible, and is accompanied by the gain or loss USA of Polycomb repressive complex (PRC)-mediated The hedgehog signaling pathway plays a central histone H3 lysine 27 trimethylation (H3K27me3) and critical role in embryonal development, and has marks in ~5.9 % of genes. These H3K27me3 also been found to play an early critical role in targets included genes associated with transcription several tumor types. Inappropriate activation of factor activity and ion channel activity pathways as the hedgehog signaling pathway in humans is well as WNT signaling and TGF-beta pathways. associated with a number of syndromes, including By contrast, no reproducible changes in CpG island

86 DNA methylation were detected. Dynamic morphologic conversion and impaired their alteration in the subcellular localization of EZH2, a tumorigenicity in NOD-SCID mice. These results catalytic component of PRC, was observed during establish the idea that epigenetic plasticity regulated the conversion of GICs. Localization of EZH2 by PRC is a key mechanism underlying adaptation was closely associated with tumor cell of tumors to their environment. PRC-targeted differentiation and plasticity in human glioblastoma. therapy reduces tumor cell plasticity and provides a Inhibition of EZH2, a PRC component that is new paradigm in cancer treatment. highly expressed in glioblastoma, disrupted the

87 Author index for research reports and publications ______An, B. J165 Ito, H. (EP) 6, 7, 8, 9, 10, 11, 22, J021, J050, J057, J058, J077, J078, Akatsuka, Y. 31, 32, J004, J044, J075, J084, J090, J114, J130, J145, J219, J222, A001, A086 J146, J147, J165, J168, J169, Aoki, M. 42, 44, J006, J017, J026, J175, J182, J187, J208, J216, J086, J107, J191 J220, R005, A012, A019, Chiba, S. J075, J135, J177 A020, A027, A028, A066, Chihara, D. 7, 10, J015, J097, J099 A067, A068 Ito, H. (MO) J024, A002 Demachi- 30, 31, J044, J083 Okamura, A. Ito, Y. 17, J055, J080, J083, J129, Fujii, M. 17, 21, 22, J003, J005, J024, J134 J074, J084, J119, J125, J128, Iwata, H. 7, 11, J011, J012, J025, J101, J162, J165, J193, A002, A015, J117, A020 A050, A051, A052, A061, Izawa, I. 47, 48, J067, A014 A064, A069, A072, A073 Kanda, T. 36, 39, J075, J076, J091, Fujita, M. 30, 32, 33, J027, J028, J173, J135, J136, J137, J151, J160, J237, J238, R001, A003, J177, J195, J230, A016, A021, A004, A005, A006, A007, A022, A023, A029, A053, A008, A032, A033, A034, A059, A060 A035, A036, A062 Kannagi, R. 42, 43, J018, J025, J065, Goto, H. 46, 47, 48, J063, J064, J067, J068, J092, J206, J217, J233 J094, J115, J123, A009, A049 Karube, K. 25, J093, J104, J197, A024, Hanai, N. 9, A028 A025, A026 Hasegawa, Y. 7, 8, 9, 17, J119, J168, J175, Kasahara, K. 46, 47, J008, J094, J115, A027, A028 J123, A009, A049 Hatooka, S. 8, J073, J168, J175, J181, Kawaguchi, K. A052 A028 Kawakita, D. 7, 9, J175, A027, A028 Hayashi, Y. 48, J063, J067 Kawamoto, E. 47 He, D. 48 Kawase, T. 7, 8, 11, J051, J057, J058, Hida, T. 7, 21, J024, J054, J128, J169, J059, J077, J101, J131, J168, J198, J232, A002, A069 J169, J175, J182, J187, J220 Hirosawa, T. 30, 31, J044, A010 Kawashima, D. J136, J137, J160, J195, A029 Honma, K. 25, 26, J093, J142, R016 Kawata, S. A052 Horio, Y. J053, J054, J128, A069 Kobori, K. 48 Hosono, S. 7, 8, 9, 10, 11, J021, J042, Kojima, Y. J017 J056, J057, J058, J059, J077, J085, J090, J101, J116, J118, Kondo, E. 15, 16, 17, J009, J024, J069, J145, J146, J147, J155, J168, J081, J082, J100, J119, J120, J169, J175, J182, J187, J220, J129, J150, J153, J154, J164, A012, A020, A027, A028, J172, J227, J228, A037, A057 A066, A067, A068, A078 Kondo, S. 30, J066 Inagaki, M. 46, 47, 48, J008, J040, J063, Kondo, Y. 21, 22, J003, J013, J023, J064, J067, J094, J101, J115, J024, J074, J084, J105, J106, J123, A009, A013 J112, J125, J127, J128, J136, Inoko, A. 48, J063, J067, A014 J140, J154, J162, J165, J180, Ishiguro, F. 21, J073, J074, J125, J128, R003, R004, R008, R010, A015, A050, A051, A052, A002, A015, A038, A039, A069 A040, A041, A042, A043, Islam, T. 7 A044, A050, A051, A052, A061, A064, A069, A072, Isomura, H. 40, J075, J076, J091, J135, A073 J136, J137, J151, J160, J177, Kumimoto, H. 51 J195, R002, A016, A017, A018 Ito, A. 17 Kuzushima, K. 30, 31, J004, J044, J055,

88 J108, J109, J161, J176, J226, J039, J079, J080, J087, J119, A030, A031, A063, A086 J120, J129, J148, J167, R007, Kyogashima, J025, J068, J130, J206 A002, A056, A057 M. Nakao, M. 7, 10, J145, J146, J147 Li, P. 46, J115, A009, A045 Nakashima, Y. 25, J093 Maseki, S. 17, J119 Nakasu, S. J177 Matsudaira, Y. J024, A002 Nakata, S. J022, J032, J052 Matsuo, K. 6, 7, 8, 9, 10, 11, J003, J010, Nakayama, S. 38, J075, J135, J151, J177, J011, J012, J015, J016, J019, A082 J020, J021, J030, J031, J033, Nishikawa, E. J157, A065 J037, J039, J041, J043, J045, J046, J047, J048, J049, J050, Nishikawa, Y. J081, J082, J153 J051, J057, J058, J059, J060, Nishio, No. 30, J007, J132, J133, J134, J073, J077, J078, J088, J089, J152, J158, J159, J196, J219, J090, J095, J098, J101, J111, J223, R009, A077 J113, J117, J122, J126, J138, Nishizawa, M. 30 J143, J144, J145, J146, J147, J148, J155, J156, J163, J166, Noda, C. 36, J137, J160, A053, A059 J168, J169, J171, J174, J175, Ogasawara, M. 17 J178, J179, J181, J182, J183, J184, J185, J186, J187, J188, Okamoto, Y. 22, J003, J084, J105, J165, J189, J200, J203, J209, J211, R010, A064, A072, A073 J212, J220, J221, J231, J232, Osada, H. 21, 22, J003, J024, J054, J236, R005, R006, R012, R23, J061, J074, J084, J125, J128, A012, A019, A020, A027, J157, J165, J192, J193, J194, A028, A047, A048, A066, J225, J229, R011, A002, A067, A068 A015, A050, A051, A052, Matsuyama, M. 46, 48, J063, J067, J115, A064, A065, A069, A072, J123, A009, A049 A073 Mitsudomi, T. 7, J073, J095, J169, J192, Oze, I. 7, 8, 9, J168, J169, J175, J193, J194, J210, J229, J231, J185, J188, J208, R006, R012, J232 A028, A066, A067, A068 Mizuno, N. 10, J043, J111, J145, J146, Saito, K. 15, J170, J172 J200 Saito, S. J136, J137, J160, J195 Mizuno, T. 21, J074, J125, J128, A015, A050, A051, A052, A069 Sakuma, K. 42, J092 Morishima Y. 7, 10, 25, 31, J015, J044, Sato, F. 7, 9, J037, J175, A028 J051, J090, J093, J096, J097, J098, J099, J113, J126, J131, Sawamura, M. 7 A026, A086 Sekido, Y. 21, 22, J002, J003, J024, Murakami, H. 21, 22, J003, J024, J062, J038, J053, J054, J062, J074, J074, J084, J125, J128, J165, J084, J105, J125, J128, J157, J193, A002, A015, A050, J165, J192, J193, J194, J198, A051, A052, A064, A069, J201, J202, J210, J224, R014, A072, A073 R015, A002, A011, A015, Murakami, S. 7, 9, 17, J119 A050, A051, A052, A064, Murata, T. 36, 37, J075, J076, J091, A065, A069, A070, A071, J135, J136, J137, J151, J160, A072, A073 J177, J195, A016, A021, A053, Seto, M. 7, 10, 25, 26, 27, J070, J071, A054, A055, A059, A082 J090, J093, J096, J097, J098, Nakagawa, M. 25, 26, 27, J093, J096, J142, J099, J110, J113, J114, J124, J218, R016, A024, A084 J141, J142, J197, J213, J214, Nakamura, S. 7, 10, 16, 17, 25, J069, J070, J215, J216, J218, R016, R017, J071, J093, J097, J104, J124, A024, A025, A026, A084, J208, J227, A024, A026 A085 Nakanishi T. 7, 8, J042, J057, J058, J059, Shinjo, K. 22, J003, J084, J105, J165, A012 A064, A072, A073 Nakanishi, H. 17, J001, J024, J029, J035, Shinoda, M. 7, 8, J168, J175, J181, A028

89 Shiraishi, K. 31, J044 Tatematsu, Y. 21, J157, A065 Sueta, A. 7, 11, A020 Tsuchida, D. 17, A056, A057 Sugimoto, A. 38, J136, J137, J160, J195, Tsurumi, T. 36, 37, 38, 39, 40, J072, J075, A074, A075 J076, J091, J135, J136, J137, Suzuki T. 7, 8, J057, J058, J059, J077, J151, J160, J177, J195, R013, J090, J101, J168, J169, J175, A016, A017, A018, A021, J212, J220, J236 A022, A023, A029, A053, Taguchi, O. 30, J139, J142, J215 A054, A055, A059, A074, A075, A082 Tajima, K. 1, 7, 8, 9, 10, 11, J011, J012, Tsuzuki, S. 25, 26, 27, J093, J142, J214, J021, J030, J033, J039, J041, J215, A024, A026, A085 J045, J048, J049, J050, J057, Uemura, Y. 30, J034, J102, J103, J121, J058, J059, J077, J090, J101, J149, A046, A058, A076, A083 J117, J122, J144, J145, J146, Watanabe, M. 8, 9, 10, J021, J057, J058, J147, J148, J163, J168, J169, J077, J087, J101, J145, J146, J171, J175, J181, J182, J187, J147, J168, J169, J175, J182, J198, J209, J211, J220, J187, J220, A012, A028, A066, J239-J243, R19-R23, A012, A067, A068 A020, A028, A066, A067, Yamao, K. 7, 10, 22, J003, J021, J043, A068, A88-92 J084, J111, J145, J146, J147, Tanaka, Ha. 17, J119, J129 J165, J200 Tanaka, Hid. 6, 7, 8, 9, 10, 11, J010, J021, Yatabe, Y. 7, 8, 9, 10, 11, 21, J003, J021, J036, J037, J057, J058, J059, J024, J031, J043, J057, J061, J077, J078, J090, J101, J145, J077, J095, J096, J097, J115, J146, J147, J168, J169, J171, J145, J146, J147, J157, J168, J175, J181, J182, J187, J190, J169, J175, J182, J187, J192, J199, J204, J205, J207, J208, J193, J194, J200, J210, J229, J220, J236, R006, R018, J231, J232, A002, A009, A026 A012, A019, A020, A027, Yokoyama, T. A072 A028, A066, A067, A068, Yusa, A. 43, J025, J092, J233 A078, A079, A080 Tanaka, Hir. 48 Zhang, R. 30, J014, J234, J235, A087