Annual Report 2018
Division of Cellular Signaling
Hiroyuki Mano, Masahito Kawazu, Yuichi Shiraishi, Shinji Kohsaka, Toshihide Ueno
[Masutani group] Mitsuko Masutani, Shoji Imamichi, Yuka Sasaki
[Shiotani Group] Bunsyo Shiotani, Kiminori Kurashima
[Masuda Group] Mari Masuda, Naoko Goto
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.
The 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 coupled with next-generation sequencing technologies.
Masutani group: This research group collaborates with the Division of Boron Neutron Capture Therapy of the Exploratory Oncology Research & Clinical Trial Center (NCC-EPOC), the Department of Radiation Oncology, NCC Hospital and other clinical research groups and studies radiation oncology focusing on BNCT and also cancer chemotherapy targeting poly(ADPribosylation) and DNA damage response pathways.
Shiotani group: Our group focuses on the mechanism underlying DNA replication stress, the hallmark of cancer that is deeply involved in the process of acquiring genome instability and cancer development, aiming at developing new therapeutic strategies.
Masuda Group: Our current research is focused on the development of the Traf2- and Nck-interacting kinase (TNIK) inhibitor, NCB0846, for the treatment of colorectal cancer and synovial sarcoma with activation of Wnt signaling. In addition, we are applying our reverse-phase protein array (RPPA) platform, which was developed in-house, to the discovery of biomarkers for use in companion diagnostics as well as identifying therapeutic targets in various types of cancer.
Research activities
1) Through collaboration with the Department of Radiation Oncology of NCC Hospital, industries, preclinical evaluations of accelerator-based BNCT system introduced in NCC were studied and carried out. As biomarkers for cellular early responses of BNCT and gamma-irradiation, biological significances of particular cytokines were characterized. From comprehensive screening of radio-sensitization targets, the APOBEC3 gene was identified and further mechanistic studies indicated a role in DNA damage responses. (Masutani group)
2) Dysfunction of poly(ADP-ribose) glycohydrolase, PARG, was shown to induce a synthetic lethal effect in dual specificity phosphatase 22-deficient lung cancer cells. The optimization for measurement of a pharmacodynamic marker of PARP inhibitor was also investigated. (Masutani group)
3) ERCC1 overexpression is reported as a candidate prognostic factor and considered to cause cisplatin resistance. Monoclonal antibodies for ERCC1 were established, evaluated and optimized for immunostaining of solid cancer specimens as a collaboration with NCC Hospital and outside institutes. (Masutani group)
4) Elevated expression of ATR suppresses lethal DNA replication stress and promotes acquiring genomic instability and transformation in KrasG12V-driven lung adenocarcinoma model. (Shiotani group)
5) SMARCA4 defects-associated heterochromatin induce intrinsic DNA replication stress and vulnerability to ATR Inhibition in Lung Adenocarcinoma. (Shiotani group)
6) Using RPPA analysis, we have determined that NCB-0846 exerts potent anti-tumor activity by inducing ATM-dependent DNA damageresponse (DDR) signaling leading to G2 arrest followed by apoptosis in colorectal cancer cells. (Masuda Group)
7) Our research has revealed potent antiproliferation activity of NCB-0846 and its unique effect of downregulating expression of the MYC gene and a broad spectrum of Wnt target genes in synovial sarcoma cells. (Masuda Group)
8) Development and optimization of our RPPA platform have enabled us to analyze phosphoproteins in clinical specimens and will lead to our active participation in the International Cancer Proteogenome Consortium (ICPC) responsible for analyzing sarcomas. (Masuda Group)
9) Applied our original method (MANOmethod) to evaluate transforming activities and drug sensitivities of variants of unknown significance in a high-throughput manner to the comprehensive functional analysis of ERBB2 mutations, and discovered several mutations related to oncogenesis and drug resistance.
10) Through an exome sequencing of 150 cases of microsatellite instability-high colorectal cancers, we showed that targetable fusion kinases were frequently detected among KRAS / BRAF mutation-negative cases with MLH1 promoter methylation. In addition, we developed a simple method for detection of MLH1 promoter methylation.
Education
Research exchanges with Nagasaki University supported partnering the graduate school by holding the Department of Comprehensive Oncology in NCC. Three graduate students and two undergraduate students from Nagasaki University visited this division for research training. A young project researcher and two adjunct scientists were nurtured in the department. (Masutani group)
We have accepted three trainees from Hoshi Pharmaceutical University, Kitasato University, and Tokyo College of Biotechnology. (Shiotani Group)
A graduate student and a medical student form Keio Medical School have joined the division for research training. We have accepted two trainees from Gakushuin University. (Masuda Group)
We accepted trainees from the University of Tokyo (5) and Juntendo University (2).
Future prospects
Based on 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 2018
Journal
1. Hirano D, Hayakawa F, Yasuda T, Tange N, Yamamoto H, Kojima Y, Morishita T, Imoto N, Tsuzuki S, Mano H, Naoe T, Kiyoi H. Chromosomal translocation-mediated evasion from miRNA induces strong MEF2D fusion protein expression, causing inhibition of PAX5 transcriptional activity. Oncogene, 38:2263-2274, 2019
2. Sato K, Kawazu M, Yamamoto Y, Ueno T, Kojima S, Nagae G, Abe H, Soda M, Oga T, Kohsaka S, Sai E, Yamashita Y, Iinuma H, Fukayama M, Aburatani H, Watanabe T, Mano H. Fusion Kinases Identified by Genomic Analyses of Sporadic Microsatellite Instability-High Colorectal Cancers. Clin Cancer Res, 25:378-389, 2019
3. Kinoshita Y, Takashi Y, Ito N, Ikegawa S, Mano H, Ushiku T, Fukayama M, Nangaku M, Fukumoto S. Ectopic expression of Klotho in fibroblast growth factor 23 (FGF23)-producing tumors that cause tumor-induced rickets/osteomalacia (TIO). Bone Rep, 10:100192, 2019
4. Watanabe K, Kage H, Shinozaki-Ushiku A, Kohsaka S, Takai D, Nakajima J, Miyagawa K, Aburatani H, Mano H, Nagase T. Spontaneous Transdifferentiation from Small Cell Lung Carcinoma to Squamous Cell Carcinoma. J Thorac Oncol, 14:e31-e34, 2019
5. Namba S, Sato K, Kojima S, Ueno T, Yamamoto Y, Tanaka Y, Inoue S, Nagae G, Iinuma H, Hazama S, Ishihara S, Aburatani H, Mano H, Kawazu M. Differential regulation of CpG island methylation within divergent and unidirectional promoters in colorectal cancer. Cancer Sci, 110:1096-1104, 2019
6. Kohsaka S, Tatsuno K, Ueno T, Nagano M, Shinozaki-Ushiku A, Ushiku T, Takai D, Ikegami M, Kobayashi H, Kage H, Ando M, Hata K, Ueda H, Yamamoto S, Kojima S, Oseto K, Akaike K, Suehara Y, Hayashi T, Saito T, Takahashi F, Takahashi K, Takamochi K, Suzuki K, Nagayama S, Oda Y, Mimori K, Ishihara S, Yatomi Y, Nagase T, Nakajima J, Tanaka S, Fukayama M, Oda K, Nangaku M, Miyazono K, Miyagawa K, Aburatani H, Mano H. Comprehensive assay for the molecular profiling of cancer by target enrichment from formalin-fixed paraffin-embedded specimens. Cancer Sci, 110:1464-1479, 2019
7. Kohsaka S, Petronczki M, Solca F, Maemondo M. Tumor clonality and resistance mechanisms in EGFR mutation-positive nonsmall-cell lung cancer: implications for therapeutic sequencing. Future Oncol, 15:637-652, 2019
8. Masuda M. Hunting hidden pieces of signaling pathways in hepatocellular carcinoma. Hepatobiliary Surg Nutr, 8:74-76, 2019
9. Nakamura S, Igaki H, Okamoto H, Wakita A, Ito M, Imamichi S, Nishioka S, Iijima K, Nakayama H, Takemori M, Kobayashi K, Abe Y, Okuma K, Takahashi K, Inaba K, Murakami N, Nakayama Y, Nishio T, Masutani M, Itami J. Dependence of neutrons generated by 7Li(p,n) reaction on Li thickness under free-air condition in accelerator-based boron neutron capture therapy system employing solid-state Li target. Phys Med, 58:121-130, 2019
10. Murakami N, Okuno Y, Yoshida K, Shiraishi Y, Nagae G, Suzuki K, Narita A, Sakaguchi H, Kawashima N, Wang X, Xu Y, Chiba K, Tanaka H, Hama A, Sanada M, Ito M, Hirayama M, Watanabe A, Ueno T, Kojima S, Aburatani H, Mano H, Miyano S, Ogawa S, Takahashi Y, Muramatsu H. Integrated molecular profiling of juvenile myelomonocytic leukemia. Blood, 131:1576-1586, 2018
11. Takahashi K, Ehata S, Koinuma D, Morishita Y, Soda M, Mano H, Miyazono K. Pancreatic tumor microenvironment confers highly malignant properties on pancreatic cancer cells. Oncogene, 37:2757-2772, 2018
12. Takashima K, Maru Y, Mori S, Mano H, Noda T, Muto K. Ethical concerns on sharing genomic data including patients' family members. BMC Med Ethics, 19:61, 2018
13. Kage H, Kohsaka S, Shinozaki-Ushiku A, Ushiku T, Takai D, Nakajima J, Miyagawa K, Aburatani H, Mano H, Nagase T. Spontaneous Transformation from EGFR and ALK Wild-Type Lung Adenocarcinoma to Neuroendocrine Carcinoma. J Thorac Oncol, 13:e126-e128, 2018
14. Asada S, Goyama S, Inoue D, Shikata S, Takeda R, Fukushima T, Yonezawa T, Fujino T, Hayashi Y, Kawabata KC, Fukuyama T, Tanaka Y, Yokoyama A, Yamazaki S, Kozuka-Hata H, Oyama M, Kojima S, Kawazu M, Mano H, Kitamura T. Mutant ASXL1 cooperates with BAP1 to promote myeloid leukaemogenesis. Nat Commun, 9:2733, 2018
15. Nagano M, Kohsaka S, Ueno T, Kojima S, Saka K, Iwase H, Kawazu M, Mano H. High-Throughput Functional Evaluation of Variants of Unknown Significance in ERBB2. Clin Cancer Res, 24:5112-5122, 2018
16. Tanaka Y, Kawazu M, Yasuda T, Tamura M, Hayakawa F, Kojima S, Ueno T, Kiyoi H, Naoe T, Mano H. Transcriptional activities of DUX4 fusions in B-cell acute lymphoblastic leukemia. Haematologica, 103:e522-e526, 2018
17. Matsuo H, Yoshida K, Fukumura K, Nakatani K, Noguchi Y, Takasaki S, Noura M, Shiozawa Y, Shiraishi Y, Chiba K, Tanaka H, Okada A, Nannya Y, Takeda J, Ueno H, Shiba N, Yamato G, Handa H, Ono Y, Hiramoto N, Ishikawa T, Usuki K, Ishiyama K, Miyawaki S, Itonaga H, Miyazaki Y, Kawamura M, Yamaguchi H, Kiyokawa N, Tomizawa D, Taga T, Tawa A, Hayashi Y, Mano H, Miyano S, Kamikubo Y, Ogawa S, Adachi S. Recurrent CCND3 mutations in MLL-rearranged acute myeloid leukemia. Blood Adv, 2:2879-2889, 2018
18. Morita K, Hama Y, Izume T, Tamura N, Ueno T, Yamashita Y, Sakamaki Y, Mimura K, Morishita H, Shihoya W, Nureki O, Mano H, Mizushima N. Genome-wide CRISPR screen identifies TMEM41B as a gene required for autophagosome formation. J Cell Biol, 217:3817-3828, 2018
19. Li JF, Dai YT, Lilljebjorn H, Shen SH, Cui BW, Bai L, Liu YF, Qian MX, Kubota Y, Kiyoi H, Matsumura I, Miyazaki Y, Olsson L, Tan AM, Ariffin H, Chen J, Takita J, Yasuda T, Mano H, Johansson B, Yang JJ, Yeoh AE, Hayakawa F, Chen Z, Pui CH, Fioretos T, Chen SJ, Huang JY. Transcriptional landscape of B cell precursor acute lymphoblastic leukemia based on an international study of 1,223 cases. Proc Natl Acad Sci USA, 115:E11711-E11720, 2018
20. Kang C, Saso K, Ota K, Kawazu M, Ueda T, Okada H. JMJD2B/ KDM4B inactivation in adipose tissues accelerates obesity and systemic metabolic abnormalities. Genes Cells, 23:767-777, 2018
21. Ishii M, Suehara Y, Sano K, Kohsaka S, Hayashi T, Kazuno S, Akaike K, Mukaihara K, Kim Y, Okubo T, Takamochi K, Takahashi F, Kaneko K, Saito T. Proteomic signatures corresponding to the SS18/SSX fusion gene in synovial sarcoma. Oncotarget, 9:37509-37519, 2018
22. Yachi K, Tsuda M, Kohsaka S, Wang L, Oda Y, Tanikawa S, Ohba Y, Tanaka S. miR-23a promotes invasion of glioblastoma via HOXD10-regulated glial-mesenchymal transition. Signal Transduct Target Ther, 3:33, 2018
23. Konishi Y, Kawamata F, Nishihara H, Homma S, Kato Y, Tsuda M, Kohsaka S, Einama T, Liu C, Yoshida T, Nagatsu A, Tanino M, Tanaka S, Kawamura H, Kamiyama T, Taketomi A. Tumor budding and human chorionic gonadotropin-beta expression correlate with unfavorable patient outcome in colorectal carcinoma. Med Oncol, 35:104, 2018
24. Akaike K, Suehara Y, Kohsaka S, Hayashi T, Tanabe Y, Kazuno S, Mukaihara K, Toda-Ishii M, Kurihara T, Kim Y, Okubo T, Hayashi Y, Takamochi K, Takahashi F, Kaneko K, Ladanyi M, Saito T. PPP2R1A regulated by PAX3/FOXO1 fusion contributes to the acquisition of aggressive behavior in PAX3/FOXO1-positive alveolar rhabdomyosarcoma. Oncotarget, 9:25206-25215, 2018
25. Yasuhara T, Kato R, Hagiwara Y, Shiotani B, Yamauchi M, Nakada S, Shibata A, Miyagawa K. Human Rad52 Promotes XPG-Mediated R-loop Processing to Initiate Transcription-Associated Homologous Recombination Repair. Cell, 175:558-570.e11, 2018
26. Nozaki T, Sasaki Y, Fukuda I, Isumi M, Nakamoto K, Onodera T, Masutani M. Next-generation sequencing-based miRNA expression analysis in Parp1-deficient embryonic stem cell-derived exosomes. Biochem Biophys Res Commun, 499:410-415, 2018
27. Nozaki T, Masutani M. p53-dependent cell cycle checkpoint after DNA damage and its relevance to PARP1. Res Rev Insights, 2:1-5, 2018
28. Nozaki T, Masutani M. Involvement of Parp1 in the downstream of p53 dependent signaling pathway induced after gamma-irradiation. J Transl Sci, 5:1-5, 2018
29. Kudo Y, Sasaki Y, Onodera T, Hashimoto J, Nozaki T, Tamura K, Watanabe W, Masutani M. Measurement of poly(ADP-ribose) level with enhanced slot blot assay with crosslinking. Challenges, 9:27, 2018
30. Miki S, Imamichi S, Fujimori H, Tomiyama A, Fujimoto K, Satomi K, Matsushita Y, Matsuzaki S, Takahashi M, Ishikawa E, Yamamoto T, Matsumura A, Mukasa A, Nishikawa R, Masutomi K, Narita Y, Masutani M, Ichimura K. Concomitant administration of radiation with eribulin improves the survival of mice harboring intracerebral glioblastoma. Cancer Sci, 109:2275-2285, 2018