Annual Report 2019
Division of Cellular Signaling
Hiroyuki Mano, Yuichi Shiraishi, Masahito Kawazu, Shinji Kohsaka, Toshihide Ueno, Satoshi Inoue,
[Masutani group] Mitsuko Masutani, Shoji Imamichi, Yuka Sasaki
[Masuda group] Mari Masuda, Naoko Goto
[Shiotani group] Bunsyo Shiotani, Kiminori Kurashima
[Yamamoto group]Yusuke Yamamoto, Iwao Shimomura, Minami Kumazaki, Hiroko Tadokoro, Anna Sanchez Calle, Lee Chuen Liew
Introduction
By sequencing the genome of cancer specimens, not only will the essential cause of carcinogenesis be clarified, but also significant progress will be made in elucidating the tumor diversity and the progression mechanism of cancer. By combining these pieces of information and functional assays, we strive to achieve the total cure of cancer patients.
The Team and What We Do
We aim to identify essential growth drivers in every cancer and develop new molecularly-targeted therapy, by taking advantage of our functional screening system coupled with the various technologies in genomics.
Masutani group: This research group collaborates with the Department of Radiation Oncology, NCC Hospital, the Division of Boron Neutron Capture Therapy of the Exploratory Oncology Research & Clinical Trial Center (NCC-EPOC), and other research institutes and studies radiation oncology focusing on BNCT and cancer chemotherapy targeting poly (ADP-ribosylation) and DNA damage response pathways.
Masuda Group: Our group focuses on developing a Traf2- and Nck-interacting kinase (TNIK) inhibitor NCB-0846 for the treatment of colorectal cancer and synovial sarcoma with activation of Wnt signaling in addition to identifying biomarkers and therapeutic targets for various types of cancer using our in-house reverse-phase protein array (RPPA) platform.
Shiotani group: Our group focuses on various obstacles in the DNA replication process to elucidate the mechanism of cancer development and the nature of cancer cells by DNA replication stress tolerance, and to develop new therapeutic strategies.
Yamamogo group: The Yamamoto group focuses on cancer stem cells, non-coding RNAs, the cancer microenvironment, and extracellular vesicles to elucidate the mechanisms of cancer development and stem cell maintenance, and to develop diagnostic markers and nucleic acid drugs for cancer.
Research activities
1) We conducted comprehensive molecular profiling of lung adenocarcinoma of never or light smokers to discover novel targetable mutations and prognostic biomarkers, and found a novel CD74-NRG2 fusion gene. The risk score generated by the expression of a three-gene set was identified to be a strong prognostic marker for overall survival and progression-free survival.
2) Through genomic analyses, we found recurring KRAS mutations in 40% of adenomyosis cases. Further analyses suggested that KRAS mutations may reduce progestin efficacy, and that adenomyosis and endometriosis may share molecular etiology, explaining their co-occurrence. These findings could lead to genetically-guided therapy and/or relapse risk assessment after uterine-sparing surgery.
3) Through collaboration with the Department of Radiation Oncology at NCC Hospital and industry, preclinical research, basic and translational studies of accelerator-based BNCT system introduced in the NCC and research on boron carrier drugs were performed. As biomarkers for cellular responses to neutron beam irradiation and gamma-irradiation, the biological significances of factors involved in tumor microenvironment were studied. (Masutani group)
4) Poly(ADP-ribose) metabolism is involved in radiation responses in various ways. Factors affecting anti-cancer potential targets such as poly(ADP-ribose) glycohydrolase, PARG, and PARP1 were investigated. (Masutani group)
5) As a prognostic factor for cisplatin resistance, the significance of ERCC1 overexpression was studied. Monoclonal antibodies developed for evaluation of ERCC1 level were characterized for immunostaining of cancer specimens in collaboration with NCC Hospital and outside institutes. (Masutani group)
6) In synovial sarcoma, NCB-0846 appears to down-regulate the MYC gene, subsequently leading to the down-regulation of various genes, which is likely responsible for the potent anti-proliferative effect of NCB0846. (Masuda group)
7) The FGF2/FGFR1 signaling was detected as a potential therapeutic target for malignant pleural mesothelioma (MPM), suggesting the clinical use of AZD4547 in treating MPM. (Masuda group)
8) With our improved RPPA platform, we identified the signaling pathways activated in gastric cancer cells and liver metastasis of pancreatic cancer in collaboration with Kumamoto University and NCC-EPOC, respectively. (Masuda group)
9) Using the KrasG12V lung line carcinoma model, we showed that elevated level of ATR expressing cells can tolerate lethal DNA replication stress and promote transformation by acquiring genomic instability. (Shiotani group)
10) Loss of SMARCA4, a core component of the SWI/SNF chromatin remodeling complex, was shown to be associated with increased heterochromatin and induced intrinsic DNA replication stress and susceptibility to ATR inhibition in lung adenocarcinoma. (Shiotani group)
11) We have reported molecular and cellular mechanisms of extracellular vesicles which are secreted by cancer cells and influence microenvironmental cells to support cancer cell survival. Furthermore, we have shown that retrotransposon activity is contained within the extracellular vesicles, indicating that DNA derived from cancer cells may be incorporated into other cells. In addition, we have identified proteins in the exosomes of pancreatic cancer patients and developed new biomarkers for miRNAs in the blood. (Yamamoto group)
12) We have identified non-coding RNAs that control cancer recurrence in breast cancer and have investigated the molecular mechanism. In addition, we also sought to identify microRNAs involved in drug resistance in oral cancer. (Yamamoto group)
13) We applied a stable culture system of normal epithelial cells to induce oncogenesis in vitro and transplanted them into immunodeficient animals to form tumors. Based on these systems, we tried to determine the cell-of-origin of cancers. These techniques were also used to perform drug screening against cancer cells with specific genetic mutations and to select multiple candidate compounds. (Yamamoto group)
Education
Young researchers and students were trained in the division as follows:
Five, two, and one graduate students from the Univ. of Tokyo, Juntendo Univ., and Univ. of Kyushu, respectively.
Research exchanges with Nagasaki Univ. supported partnering with the graduate school by holding the Department of Comprehensive Oncology in the NCC. A young project researcher and two adjunct scientists were trained and three graduate students and two undergraduate students from Nagasaki Univ. visited this division for research training. (Masutani group)
A graduate student and a medical student from Keio Univ. School of Medicine. A graduate student and two trainees from Gakushuin Univ. (Masuda group)
Three trainees from Hoshi Pharmaceutical Univ., a trainee from Kitasato Univ., and a trainee from Tokyo College of Biotechnology. (Shiotani group)
Three trainees from Jikei Univ. School of Medicine, and a trainee from Keio Univ. Faculty of Pharmacy. (Yamamoto group)
Future prospects
Based on recent technical improvements in sequencing technologies, implementation of clinical sequencing is being achieved. Through the development of sequencing methods coupled with functional screening systems, and the identification of useful biomarkers, we aim to contribute to the improvement and implementation of precision medicine.
List of papers published in 2019
Journal
1. Aizawa H, Karasaki T, Nagayama K, Shinozaki-Ushiku A, Aburatani H, Mano H, Nakajima J. Clinical Application of Next-generation Sequencing for the Diagnosis of Lung Squamous Cell Carcinoma: Is It Primary or Secondary? Intern Med, 59:1299-1302, 2020
2. Hayakawa D, Takahashi F, Mitsuishi Y, Tajima K, Hidayat M, Winardi W, Ihara H, Kanamori K, Matsumoto N, Asao T, Ko R, Shukuya T, Takamochi K, Hayashi T, Suehara Y, Takeda Nakamura I, Ueno T, Kohsaka S, Mano H, Takahashi K. Activation of insulin-like growth factor-1 receptor confers acquired resistance to osimertinib in non-small cell lung cancer with EGFR T790M mutation. Thorac Cancer, 11:140-149, 2020
3. Kohsaka S, Hayashi T, Nagano M, Ueno T, Kojima S, Kawazu M, Shiraishi Y, Kishikawa S, Suehara Y, Takahashi F, Takahashi K, Suzuki K, Takamochi K, Mano H. Identification of Novel CD74-NRG2α Fusion From Comprehensive Profiling of Lung Adenocarcinoma in Japanese Never or Light Smokers. J Thorac Oncol, 15:948-961, 2020
4. Makise N, Mori T, Kobayashi H, Nakagawa K, Ryo E, Nakajima J, Kohsaka S, Mano H, Aburatani H, Yoshida A, Ushiku T. Mesenchymal tumours with RREB1-MRTFB fusion involving the mediastinum: extra-glossal ectomesenchymal chondromyxoid tumours? Histopathology, 76:1023-1031, 2020
5. Hayashi T, Takamochi K, Kohsaka S, Kishikawa S, Suehara Y, Takahashi F, Suzuki K, Saito T, Yao T. Transformation from EGFR/PTEN co-mutated lung adenocarcinoma to small cell carcinoma in lymph node metastasis. Pathol Int, 70:295-299, 2020
6. Miyanaga A, Masuda M, Motoi N, Tsuta K, Nakamura Y, Nishijima N, Watanabe SI, Asamura H, Tsuchida A, Seike M, Gemma A, Yamada T. Whole-exome and RNA sequencing of pulmonary carcinoid reveals chromosomal rearrangements associated with recurrence. Lung Cancer, 145:85-94, 2020
7. Sugano T, Yoshida M, Masuda M, Ono M, Tamura K, Kinoshita T, Tsuda H, Honda K, Gemma A, Yamada T. Prognostic impact of ACTN4 gene copy number alteration in hormone receptor-positive, HER2-negative, node-negative invasive breast carcinoma. Br J Cancer, 122:1811-1817, 2020
8. Yamamoto Y, Kondo S, Matsuzaki J, Esaki M, Okusaka T, Shimada K, Murakami Y, Enomoto M, Tamori A, Kato K, Aoki Y, Takizawa S, Sakamoto H, Niida S, Takeshita F, Ochiya T. Highly Sensitive Circulating MicroRNA Panel for Accurate Detection of Hepatocellular Carcinoma in Patients With Liver Disease. Hepatol Commun, 4:284-297, 2020
9. Hironaka-Mitsuhashi A, Otsuka K, Gailhouste L, Sanchez Calle A, Kumazaki M, Yamamoto Y, Fujiwara Y, Ochiya T. MiR-1285-5p/TMEM194A axis affects cell proliferation in breast cancer. Cancer Sci, 111:395-405, 2020
10. Asakura K, Kadota T, Matsuzaki J, Yoshida Y, Yamamoto Y, Nakagawa K, Takizawa S, Aoki Y, Nakamura E, Miura J, Sakamoto H, Kato K, Watanabe SI, Ochiya T. A miRNA-based diagnostic model predicts resectable lung cancer in humans with high accuracy. Commun Biol, 3:134, 2020
11. Prieto-Vila M, Ochiya T, Yamamoto Y. Single-cell qPCR Assay with Massively Parallel Microfluidic System. Bio-protocol, 10:e3563, 2020
12. Sasaki Y, Fujimori H, Hozumi M, Onodera T, Nozaki T, Murakami Y, Ashizawa K, Inoue K, Koizumi F, Masutani M. Dysfunction of Poly (ADP-Ribose) Glycohydrolase Induces a Synthetic Lethal Effect in Dual Specificity Phosphatase 22-Deficient Lung Cancer Cells. Cancer Res, 79:3851-3861, 2019
13. Chen L, Gunji A, Uemura A, Fujihara H, Nakamoto K, Onodera T, Sasaki Y, Imamichi S, Isumi M, Nozaki T, Kamada N, Jishage KI, Masutani M. Development of renal failure in PargParp-1 null and Timm23 hypomorphic mice. Biochem Pharmacol, 167:116-124, 2019
14. Fujihara H, Nozaki T, Tsutsumi M, Isumi M, Shimoda S, Hamada Y, Masutani M. Spontaneous Development of Dental Dysplasia in Aged Parp-1 Knockout Mice. Cells, 8:2019
15. Motegi A, Masutani M, Yoshioka KI, Bessho T. Aberrations in DNA repair pathways in cancer and therapeutic significances. Semin Cancer Biol, 58:29-46, 2019
16. Nakamura S, Igaki H, Ito M, Okamoto H, Nishioka S, Iijima K, Nakayama H, Takemori M, Imamichi S, Kashihara T, Takahashi K, Inaba K, Okuma K, Murakami N, Abe Y, Nakayama Y, Masutani M, Nishio T, Itami J. Characterization of the relationship between neutron production and thermal load on a target material in an accelerator-based boron neutron capture therapy system employing a solid-state Li target. PLoS One, 14:e0225587, 2019
17. 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 (7)Li(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
18. Naito Y, Yamamoto Y, Sakamoto N, Shimomura I, Kogure A, Kumazaki M, Yokoi A, Yashiro M, Kiyono T, Yanagihara K, Takahashi RU, Hirakawa K, Yasui W, Ochiya T. Cancer extracellular vesicles contribute to stromal heterogeneity by inducing chemokines in cancer-associated fibroblasts. Oncogene, 38:5566-5579, 2019
19. Inoue S, Hirota Y, Ueno T, Fukui Y, Yoshida E, Hayashi T, Kojima S, Takeyama R, Hashimoto T, Kiyono T, Ikemura M, Taguchi A, Tanaka T, Tanaka Y, Sakata S, Takeuchi K, Muraoka A, Osuka S, Saito T, Oda K, Osuga Y, Terao Y, Kawazu M, Mano H. Uterine adenomyosis is an oligoclonal disorder associated with KRAS mutations. Nat Commun, 10:5785, 2019
20. Ando M, Kobayashi H, Shinozaki-Ushiku A, Chikuda H, Matsubayashi Y, Yoshida M, Saito Y, Kohsaka S, Oda K, Miyagawa K, Aburatani H, Mano H, Yamasoba T. Spinal solitary fibrous tumor of the neck: Next-generation sequencing-based analysis of genomic aberrations. Auris Nasus Larynx, 23:S0385, 2019
21. Hasegawa N, Takeda Nakamura I, Ueno T, Kojima S, Kawazu M, Akaike K, Okubo T, Takagi T, Suehara Y, Hayashi T, Saito T, Kaneko K, Mano H, Kohsaka S. Detection of circulating sarcoma tumor cells using a microfluidic chip-type cell sorter. Sci Rep, 9:20047, 2019
22. Kage H, Kohsaka S, Shinozaki-Ushiku A, Hiraishi Y, Sato J, Nagayama K, Ushiku T, Takai D, Nakajima J, Miyagawa K, Aburatani H, Mano H, Nagase T. Small lung tumor biopsy samples are feasible for high quality targeted next generation sequencing. Cancer Sci, 110:2652-2657, 2019
23. 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
24. 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
25. Oga T, Yamashita Y, Soda M, Kojima S, Ueno T, Kawazu M, Suzuki N, Nagano H, Hazama S, Izumiya M, Koike K, Mano H. Genomic profiles of colorectal carcinoma with liver metastases and newly identified fusion genes. Cancer Sci, 110:2973-2981, 2019
26. Sai E, Miwa Y, Takeyama R, Kojima S, Ueno T, Yashiro M, Seto Y, Mano H. Identification of candidates for driver oncogenes in scirrhous-type gastric cancer cell lines. Cancer Sci, 110:2643-2651, 2019
27. Takaoka K, Kawazu M, Koya J, Yoshimi A, Masamoto Y, Maki H, Toya T, Kobayashi T, Nannya Y, Arai S, Ueno T, Ueno H, Suzuki K, Harada H, Manabe A, Hayashi Y, Mano H, Kurokawa M. A germline HLTF mutation in familial MDS induces DNA damage accumulation through impaired PCNA polyubiquitination. Leukemia, 33:1773-1782, 2019
28. Yoshimoto T, Matsubara D, Soda M, Ueno T, Amano Y, Kihara A, Sakatani T, Nakano T, Shibano T, Endo S, Hagiwara K, Fukayama M, Denda-Nagai K, Irimura T, Mano H, Niki T. Mucin 21 is a key molecule involved in the incohesive growth pattern in lung adenocarcinoma. Cancer Sci, 110:3006-3011, 2019
29. Suehara Y, Okubo T, Kurihara T, Hayashi T, Kohsaka S, Kazuno S, Sano K, Hasegawa N, Miura Y, Akaike K, Kim Y, Takamochi K, Takahashi F, Ueno T, Kaneko K, Saito T. Protein Expression Profiles Corresponding to Histological Changes with Denosumab Treatment in Giant Cell Tumors of Bone. Proteomics Clin Appl, 13:e1800147, 2019
30. Masuda M, Yamada T. Utility of Reverse-Phase Protein Array for Refining Precision Oncology. Adv Exp Med Biol, 1188:239-249, 2019
31. Xian W, Duleba M, Zhang Y, Yamamoto Y, Ho KY, Crum C, McKeon F. The Cellular Origin of Barrett's Esophagus and Its Stem Cells. Adv Exp Med Biol, 1123:55-69, 2019
32. Urabe F, Matsuzaki J, Yamamoto Y, Kimura T, Hara T, Ichikawa M, Takizawa S, Aoki Y, Niida S, Sakamoto H, Kato K, Egawa S, Fujimoto H, Ochiya T. Large-scale Circulating microRNA Profiling for the Liquid Biopsy of Prostate Cancer. Clin Cancer Res, 25:3016-3025, 2019
33. Shimomura I, Yokoi A, Kohama I, Kumazaki M, Tada Y, Tatsumi K, Ochiya T, Yamamoto Y. Drug library screen reveals benzimidazole derivatives as selective cytotoxic agents for KRAS-mutant lung cancer. Cancer Lett, 451:11-22, 2019
34. Sonoda T, Matsuzaki J, Yamamoto Y, Sakurai T, Aoki Y, Takizawa S, Niida S, Ochiya T. Serum MicroRNA-Based Risk Prediction for Stroke. Stroke, 50:1510-1518, 2019
35. Zhou Y, Yamamoto Y, Xiao Z, Ochiya T. The Immunomodulatory Functions of Mesenchymal Stromal/Stem Cells Mediated via Paracrine Activity. J Clin Med, 8:1025, 2019
36. Kawamura Y, Sanchez Calle A, Yamamoto Y, Sato TA, Ochiya T. Extracellular vesicles mediate the horizontal transfer of an active LINE-1 retrotransposon. J Extracell Vesicles, 8:1643214, 2019
37. Satomi-Tsushita N, Shimomura A, Matsuzaki J, Yamamoto Y, Kawauchi J, Takizawa S, Aoki Y, Sakamoto H, Kato K, Shimizu C, Ochiya T, Tamura K. Serum microRNA-based prediction of responsiveness to eribulin in metastatic breast cancer. PLoS One, 14:e0222024, 2019
38. Sato J, Shimomura A, Kawauchi J, Matsuzaki J, Yamamoto Y, Takizawa S, Sakamoto H, Ohno M, Narita Y, Ochiya T, Tamura K. Brain metastasis-related microRNAs in patients with advanced breast cancer. PLoS One, 14:e0221538, 2019
39. Yokoi A, Matsuzaki J, Yamamoto Y, Tate K, Yoneoka Y, Shimizu H, Uehara T, Ishikawa M, Takizawa S, Aoki Y, Kato K, Kato T, Ochiya T. Serum microRNA profile enables preoperative diagnosis of uterine leiomyosarcoma. Cancer Sci, 110:3718-3726, 2019
Book
1. Yamamoto Y, Kohama, Ochiya T. miRNAs and Hepatocellular Carcinoma. In: Arias IM, Alter HJ, Boyer JL, Cohen DE, Shafritz DA, Thorgeirsson SS, Wolkoff AW (eds), The Liver: Biology and Pathobiology, 6th Edition, USA, Wiley, pp 183-194, 2020