Annual Report 2017
Division of Cellular Signaling
Hiroyuki Mano, Masahito Kawazu, Shinji Kohsaka, Toshihide Ueno
[Masutani group] Mitsuko Masutani, Shoji Imamichi, Yuka Sasaki, Takae Onodera, Makoto Ihara, Takayuki Oishi, Yusuke Kimura
[Shiotani Group] Bunsho Shiotani, Marianne Hanae Mazevet, Kiminori Kurashima
[Masuda Group] Mari Masuda, Tesshi Yamada, Naoko Goto, Toru Hirozane, Teppei Sugano, HaruyoTozaki
Introduction
By sequencing the genome of cancer specimens, not only the essential cause of carcinogenesis will be clarified, but also significant progress will be made in elucidating the tumor diversity and the progression mechanism of cancer. By combining these information and functional assays we strive for the total cure of cancer patients.
Our team and what we do
We aim at identifying essential growth drivers in every cancer and developing new molecularly-targeted therapy, by taking advantage of our functional screening system coupledwith the next generation sequencing technologies. Masutani group: This research group collaborates with the Division of Boron Neutron Capture Therapy (BNCT) of the Exploratory Oncology Research & Clinical Trial Center (NCC-EPOC), the Department of Radiation Oncology of the National Cancer Center Hospital (NCCH), and other clinical departments, and studies radiation oncology focusing on BNCT and also cancer chemotherapy targeting poly (ADP-ribosylation) and DNA damage response pathways.
Shiotani group: This group focuses on the mechanism underlying DNA replication stress response that is deeply involved in the process of cancer development and the hallmark of cancer cells aiming at developing new therapeutic strategies.
Masuda group: This group's current research is directed at the development of a novel Traf2- and Nck-interacting kinase (TNIK) inhibitors against colorectal cancer with activation of Wnt signaling. We have also applied our in-house reverse-phase protein array (RPPA) platform to the discovery of biomarkers for companion diagnostics as well as therapeutic targets in various types of cancer.
Research activities
1) By collaboration with the Department of Radiation Oncology of the NCCH, and industries, strategies and conditions for preclinical evaluation of accelerator-based BNCT system introduced in the NCC was investigated. As a biomarker for cellular early responses of BNCT and gamma-irradiation, GM-CSF was found and the biological significances were characterized. From comprehensive screening of radio-sensitization targets, IL27 receptor was identified and mechanistic studies indicated its role in controlling inflammatory responses after gamma-irradiation. (Masutani group)
2) As a biomarker for cisplatin treatment, a monoclonal antibody for ERCC1 to detect its overexpression was established and the immunostaining condition was optimized for gastric cancer specimen in collaboration with the NCCH and outside institutes. (Masutani group)
3) Measurement conditions for a pharmacodynamic marker of PARP inhibitors were investigated and improved. Candidate predictive biomarkers for PARP inhibitors for blood cancer were also investigated with outside institutes in basic studies. (Masutani group)
4) We identified that elevated expression of ATR responding to DNA replication stress suppresses lethal DNA replication stress and promotes transformation in a KrasG12V driven lung adenocarcinoma model. (Shiotani group)
5) We identified that DNA replication stress level of lung adenocarcinoma cells strongly correlates with ATR inhibitor susceptibility. (Shiotani group)
6) In collaboration with Carna Biosciences, Inc., Kobe, Japan, we synthesized 285 derivatives of the TNIK inhibitor NCB-0846 and developed a high-throughput cell-based assay for Wnt signaling activity to screen these compounds for one with the highest selectivity and lowest toxicity. (Masuda group)7) We found that NCB-0846 exerts potent anti- tumor activity in an osteosarcoma (OS) xenografted model by inducing adipocyte transdifferentiation of OS cells, and that it also blocks the TGFß-triggered epithelial mesenchymal transition (EMT) of lung cancer cells, suggesting its potential application to clinical management of distant metastasis of lung cancer. (Masuda group)
8) We established and optimized a reverse-phase protein array platform (RPPA) for analysis of phosphoproteins in clinical specimens, and served as a member of the International Cancer Proteogenome Cosortium (ICPC) responsible for analysis of sarcomas. (Masuda group)
9) We developed a novel method (MANO-method) to evaluate transforming activities and drug sensitivities of variants of unknown significance in a high-throughput manner, applied to the comprehensive functional analysis of EGFR mutations, and discovered several mutations related to oncogenesis and drug resistance.
10) Through the whole genome sequencing of triple negative breast cancer, we observed frequent silencing of BRCA1 in tumors from Japanese population, where the genomic structural variations caused by homologous recombination deficiency were highly accumulated.
Education
We accepted trainees from The University of Tokyo (3), Juntendo University (1), Keio Medical School (1), Nippon Medical School (1), and Chiba University (2). Research exchanges with Nagasaki University supported partnering the graduate school by opening the Department of Comprehensive Oncology in the NCC. Three graduate students and two undergraduate students from Nagasaki University visited our division for research training.
Future prospects
On the basis of the recent technical improvement of sequencing technologies, implementation of clinical sequencing is being realized. Through the development of sequencing methods coupled with functional screening systems, and identification of useful biomarkers, we aim to contribute to the improvement and implementation of precision medicine.
List of papers published in January 2017 - March 2018
Journal
1. Kawazu M, Kojima S, Ueno T, Totoki Y, Nakamura H, Kunita A, Qu W, Yoshimura J, Soda M, Yasuda T, Hama N, Saito-Adachi M, Sato K, Kohsaka S, Sai E, Ikemura M, Yamamoto S, Ogawa T, Fukayama M, Tada K, Seto Y, Morishita S, Hazama S, Shibata T, Yamashita Y, Mano H. Integrative analysis of genomic alterations in triple-negative breast cancer in association with homologous recombination deficiency. PLoS Genet, 13:e1006853, 2017
2. Piao J, Takai S, Kamiya T, Inukai T, Sugita K, Ohyashiki K, Delia D, Masutani M, Mizutani S, Takagi M. Poly (ADP-ribose) polymerase inhibitors selectively induce cytotoxicity in TCF3-HLF-positive leukemic cells. Cancer Lett, 386:131-140, 2017
3. Yamada T, Masuda M. Emergence of TNIK inhibitors in cancer therapeutics. Cancer Sci, 108:818-823, 2017
4. Seki M, Kimura S, Isobe T, Yoshida K, Ueno H, Nakajima-Takagi Y, Wang C, Lin L, Kon A, Suzuki H, Shiozawa Y, Kataoka K, Fujii Y, Shiraishi Y, Chiba K, Tanaka H, Shimamura T, Masuda K, Kawamoto H, Ohki K, Kato M, Arakawa Y, Koh K, Hanada R, Moritake H, Akiyama M, Kobayashi R, Deguchi T, Hashii Y, Imamura T, Sato A, Kiyokawa N, Oka A, Hayashi Y, Takagi M, Manabe A, Ohara A, Horibe K, Sanada M, Iwama A, Mano H, Miyano S, Ogawa S, Takita J. Recurrent SPI1 (PU.1) fusions in high-risk pediatric T cell acute lymphoblastic leukemia. Nat Genet, 49:1274-1281, 2017
5. Kohsaka S, Nagano M, Ueno T, Suehara Y, Hayashi T, Shimada N, Takahashi K, Suzuki K, Takamochi K, Takahashi F, Mano H. A method of high-throughput functional evaluation of EGFR gene variants of unknown significance in cancer. Sci Transl Med, 9:2017
6. Masuda M, Yamada T. The emergence of TNIK as a therapeutic target for colorectal cancer. Expert Opin Ther Targets, 21:353-355, 2017
7. Takamochi K, Takahashi F, Suehara Y, Sato E, Kohsaka S, Hayashi T, Kitano S, Uneno T, Kojima S, Takeuchi K, Mano H, Suzuki K. DNA mismatch repair deficiency in surgically resected lung adenocarcinoma: Microsatellite instability analysis using the Promega panel. Lung Cancer, 110:26-31, 2017
8. Matsubara D, Soda M, Yoshimoto T, Amano Y, Sakuma Y, Yamato A, Ueno T, Kojima S, Shibano T, Hosono Y, Kawazu M, Yamashita Y, Endo S, Hagiwara K, Fukayama M, Takahashi T, Mano H, Niki T. Inactivating mutations and hypermethylation of the NKX2-1/TTF-1 gene in non-terminal respiratory unit-type lung adenocarcinomas. Cancer Sci, 108:1888-1896, 2017
9. Masuda M, Yamada T. Signaling pathway profiling using reverse-phase protein array and its clinical applications. Expert Rev Proteomics, 14:607-615, 2017
10. Shoji H, Tada K, Kitano S, Nishimura T, Shimada Y, Nagashima K, Aoki K, Hiraoka N, Honma Y, Iwasa S, Takashima A, Kato K, Boku N, Honda K, Yamada T, Heike Y, Hamaguchi T. The peripheral immune status of granulocytic myeloid-derived suppressor cells correlates the survival in advanced gastric cancer patients receiving cisplatin-based chemotherapy. Oncotarget, 8:95083-95094, 2017
11. Nakamura S, Imamichi S, Masumoto K, Ito M, Wakita A, Okamoto H, Nishioka S, Iijima K, Kobayashi K, Abe Y, Igaki H, Kurita K, Nishio T, Masutani M, Itami J. Evaluation of radioactivity in the bodies of mice induced by neutron exposure from an epi-thermal neutron source of an accelerator-based boron neutron capture therapy system. Proc Jpn Acad Ser B Phys Biol Sci, 93:821-831, 2017
12. Nakamura T, Yamashita S, Fukumura K, Nakabayashi J, Tanaka K, Tamura K, Tateishi K, Kinoshita M, Fukushima S, Takami H, Fukuoka K, Yamazaki K, Matsushita Y, Ohno M, Miyakita Y, Shibui S, Kubo A, Shuto T, Kocialkowski S, Yamanaka S, Mukasa A, Sasayama T, Mishima K, Maehara T, Kawahara N, Nagane M, Narita Y, Mano H, Ushijima T, Ichimura K. Genome-wide DNA methylation profiling identifies primary central nervous system lymphoma as a distinct entity different from systemic diffuse large B-cell lymphoma. Acta Neuropathol, 133:321-324, 2017
13. Soh SX, Siddiqui FJ, Allen JC, Kim GW, Lee JC, Yatabe Y, Soda M, Mano H, Soo RA, Chin TM, Ebi H, Yano S, Matsuo K, Niu X, Lu S, Isobe K, Lee JH, Yang JC, Zhao M, Zhou C, Lee JK, Lee SH, Lee JY, Ahn MJ, Tan TJ, Tan DS, Tan EH, Ong ST, Lim WT. A systematic review and meta-analysis of individual patient data on the impact of the BIM deletion polymorphism on treatment outcomes in epidermal growth factor receptor mutant lung cancer. Oncotarget, 8:41474-41486, 2017
14. Hashimoto T, Yamashita S, Yoshida H, Taniguchi H, Ushijima T, Yamada T, Saito Y, Ochiai A, Sekine S, Hiraoka N. WNT Pathway Gene Mutations Are Associated With the Presence of Dysplasia in Colorectal Sessile Serrated Adenoma/Polyps. Am J Surg Pathol, 41:1188-1197, 2017
15. Mukaihara K, Tanabe Y, Kubota D, Akaike K, Hayashi T, Mogushi K, Hosoya M, Sato S, Kobayashi E, Okubo T, Kim Y, Kohsaka S, Saito T, Kaneko K, Suehara Y. Cabozantinib and dastinib exert anti-tumor activity in alveolar soft part sarcoma. PLoS One, 12:e0185321, 2017
16. Hirata Y, Kobayashi T, Nishiumi S, Yamanaka K, Nakagawa T, Fujigaki S, Iemoto T, Kobayashi M, Okusaka T, Nakamori S, Shimahara M, Ueno T, Tsuchida A, Sata N, Ioka T, Yasunami Y, Kosuge T, Kaneda T, Kato T, Yagihara K, Fujita S, Yamada T, Honda K, Azuma T, Yoshida M. Identification of highly sensitive biomarkers that can aid the early detection of pancreatic cancer using GC/MS/MS-based targeted metabolomics. Clin Chim Acta, 468:98-104, 2017
17. Shiraishi H, Fujiwara Y, Kakuya T, Tsuta K, Motoi N, Miura N, Watabe Y, Watanabe SI, Noro R, Nagashima K, Huang W, Yamada T, Asamura H, Ohe Y, Honda K. Actinin-4 protein overexpression as a predictive biomarker in adjuvant chemotherapy for resected lung adenocarcinoma. Biomark Med, 10.2217/bmm-2017-0150, 2017
18. Shimura T, Tada Y, Hirai H, Kawakita D, Kano S, Tsukahara K, Shimizu A, Takase S, Imanishi Y, Ozawa H, Okami K, Sato Y, Fushimi C, Takahashi H, Okada T, Sato H, Otsuka K, Watanabe Y, Sakai A, Ebisumoto K, Togashi T, Ueki Y, Ota H, Ando M, Kohsaka S, Hanazawa T, Chazono H, Kadokura Y, Kobayashi H, Nagao T. Prognostic and histogenetic roles of gene alteration and the expression of key potentially actionable targets in salivary duct carcinomas. Oncotarget, 9:1852-1867, 2018
19. Tanabe Y, Suehara Y, Kohsaka S, Hayashi T, Akaike K, Mukaihara K, Kurihara T, Kim Y, Okubo T, Ishii M, Kazuno S, Kaneko K, Saito T. IRE1alpha-XBP1 inhibitors exerted anti-tumor activities in Ewing's sarcoma. Oncotarget, 9:14428-14443, 2018
20. Komuro A, Raja E, Iwata C, Soda M, Isogaya K, Yuki K, Ino Y, Morikawa M, Todo T, Aburatani H, Suzuki H, Ranjit M, Natsume A, Mukasa A, Saito N, Okada H, Mano H, Miyazono K, Koinuma D. Identification of a novel fusion gene HMGA2-EGFR in glioblastoma. Int J Cancer, 142:1627-1639, 2018
21. Kohsaka S, Saito T, Akaike K, Suehara Y, Hayashi T, Takagi T, Kaneko K, Ueno T, Kojima S, Kohashi KI, Mano H, Oda Y, Yao T. Pediatric soft tissue tumor of the upper arm with LMNA-NTRK1 fusion. Hum Pathol, 72:167-173, 2018
22. Kawamata F, Nishihara H, Homma S, Kato Y, Tsuda M, Konishi Y, Wang L, Kohsaka S, Liu C, Yoshida T, Tanino M, Tanaka S, Kawamura H, Kamiyama T, Taketomi A. Chorionic Gonadotropin-beta Modulates Epithelial-Mesenchymal Transition in Colorectal Carcinoma Metastasis. Am J Pathol, 188:204-215, 2018