Annual Report 2021

Division of Cellular Signaling

Hiroyuki Mano, Masahito Kawazu, Shinji Kohsaka, Toshihide Ueno, Yosuke Tanaka

Introduction

 Through an approach combining high-sensitivity functional screening with next-generation sequencing analysis, we aim to elucidate the development mechanisms of human tumors and develop novel molecular targeted therapies.

 The Team and What We Do

• Identification of tumorigenesis and development of new drugs through comprehensive genome analysis.
• Establishment of functional assays to evaluate gene alterations.
• Development of new computational pipelines for cancer research.

 Research activities

1) Peritoneal metastasis, a hallmark of incurable advanced gastric cancer (GC), currently has no curative therapy and its molecular features have not been examined extensively. We presented a comprehensive multi-omic analysis of malignant ascitic fluid samples and their corresponding tumor cell lines from 98 patients, including whole-genome sequencing, RNA sequencing, DNA methylation and enhancer landscape. We identified potentially targetable gene alterations in approximately half of all cases. Our analyses also stratified ascites-disseminated GC into two distinct molecular subtypes: one displaying active super enhancers (SEs) at the ELF3, KLF5 and EHF loci, and a second subtype bearing TGF-β pathway activation through SMAD3 SE activation and high expression of TEAD1 (Tanaka Y et al., 2, 9, 962-977, Nat Cancer. 2021).

2) Mutations in HLA-ABC genes were analyzed in 114 microsatellite instability-high colorectal cancers using comprehensive multi-omic analysis. We detected 101 truncating mutations in 57 tumors (50%) and loss of 61 alleles in 21 tumors (18%). We identified a subtype of tumors in which lymphocyte infiltration was reduced, partly due to reduced expression of HLA-ABC genes in the absence of apparent genetic alterations. Survival of patients with such tumors was shorter than in patients with other tumor types. Various genetic and epigenetic alterations converged on reduced expression of HLA-ABC genes (Kawazu M et al., 162, 3, 799-812, Gastroenterology. 2021).

3) We performed next-generation sequencing to determine the genomic and transcriptomic profiles of pulmonary pleomorphic carcinoma (PPC). We sequenced the DNAs and RNAs of 78 specimens from 52 patients with PPC and analyzed 15 PPC cases to identify intratumoral differences between epithelial and sarcomatoid components. Epithelial and sarcomatoid components shared activating driver mutations, and there were no significant differences in CD274 expression or TMB between the two components. However, PD-L1 was highly expressed in the sarcomatoid component of several cases compared with the epithelial component. Our data suggest that therapies targeting activating driver mutations may be effective for patients with PPC and that immune checkpoint inhibitors of PPC may be recommended after careful assessment of PD-L1 expression in each epithelial and sarcomatoid component (Nagano M et al., 5, 1, 57, NPJ Precis Oncol. 2021).

4) We evaluated the transforming activity and sensitivity of 160 nonsynonymous FGFR mutations and ten fusion genes to seven FGFR tyrosine kinase inhibitors (TKIs) using the mixed-all-nominated-in-one (MANO) method, a high-throughput functional assay. The oncogenicity of 71 mutants was newly discovered in this study. The FGFR TKIs showed anti-proliferative activities against the wild-type FGFRs and their fusions, while several hotspot mutants were relatively resistant to those TKIs. Comprehensive analysis of published FGFR structures revealed a possible mechanism through which oncogenic FGFR mutations reduce sensitivity to TKIs. Our study suggests the importance of selecting suitable inhibitors against individual FGFR variants. Moreover, it reveals the necessity to develop next-generation FGFR inhibitors that are effective against all oncogenic FGFR variants (Nakamura IT et al., 5, 1, 66, NPJ Precis Oncol. 2021).

5) The clinical sequencing of tumors is usually performed on formalin-fixed, paraffin-embedded (FFPE) samples and results in many sequencing errors. We identified that most of these errors are detected in chimeric reads caused by single-strand DNA molecules with microhomology. During the end-repair step of library preparation, mutations are introduced by the mis-annealing of two single-strand DNA molecules comprising homologous sequences. The mutated bases are distributed unevenly near the ends in the individual reads. Our filtering pipeline, MicroSEC, focuses on the uneven distribution of mutations in each read and removes the sequencing errors in FFPE samples without over-eliminating the mutations detected also in fresh frozen samples. Our pipeline will increase the reliability of the clinical sequencing and advance the cancer research using FFPE samples (Ikegami M et al., 4, 1, 1396, Commun Biol. 2021).

6) We evaluated the drug sensitivity of various EGFR mutants to monotherapies and combination therapies of EGFR-tyrosine kinase inhibitors (TKIs). In vitro, the transforming potential and drug sensitivity of 357 EGFR variants were assessed. In vivo, we tested the sensitivity of EGFR variants to different regimens of EGFR-TKIs by examining changes in the proportion of each variant within the tumor. Despite the lack of a synergistic impact, TKI combination treatment effectively reduced in vivo the heterogeneous tumors composed of 3T3 cells with different EGFR variants. Combination EGFR-TKI treatment may decrease tumor growth and prevent the development of resistant variants. This work created an experimental model of a heterogeneous tumor to find the best combination therapy regimen and proposed a basic notion of EGFR-TKI combination therapy to enhance the prognosis of NSCLC patients (Ikeuchi H et al., 41, 17, 2470-2479, Oncogene. 2022).

Education

 Four trainees from The Univ. of Tokyo, one from Kyushu Univ., three from Juntendo Univ., one from Jikei Univ., and one from Yokohama City Univ.

The Future

 Clinical genomics is now being implemented in clinical practice, and sequencing technologies including whole genome sequencing, long read sequencing, and single cell analysis are advancing remarkably. We aim to contribute to the development and further improvement of clinical medicine through research and development of sequencing and screening technologies, and identification of useful biomarkers.

List of papers published in 2021

Journal

1. Jo H, Yagishita S, Hayashi Y, Ryu S, Suzuki M, Kohsaka S, Ueno T, Matsumoto Y, Horinouchi H, Ohe Y, Watanabe SI, Motoi N, Yatabe Y, Mano H, Takahashi K, Hamada A. Comparative Study on the Efficacy and Exposure of Molecular Target Agents in Non-small Cell Lung Cancer PDX Models with Driver Genetic Alterations. Molecular cancer therapeutics, 21:359-370, 2022

2. Kumagai S, Koyama S, Itahashi K, Tanegashima T, Lin YT, Togashi Y, Kamada T, Irie T, Okumura G, Kono H, Ito D, Fujii R, Watanabe S, Sai A, Fukuoka S, Sugiyama E, Watanabe G, Owari T, Nishinakamura H, Sugiyama D, Maeda Y, Kawazoe A, Yukami H, Chida K, Ohara Y, Yoshida T, Shinno Y, Takeyasu Y, Shirasawa M, Nakama K, Aokage K, Suzuki J, Ishii G, Kuwata T, Sakamoto N, Kawazu M, Ueno T, Mori T, Yamazaki N, Tsuboi M, Yatabe Y, Kinoshita T, Doi T, Shitara K, Mano H, Nishikawa H. Lactic acid promotes PD-1 expression in regulatory T cells in highly glycolytic tumor microenvironments. Cancer cell, 40:201-218.e9, 2022

3. Suzuki K, Igata H, Abe M, Yamamoto Y. Multiple cancer type classification by small RNA expression profiles with plasma samples from multiple facilities. Cancer science, 2022

4. Yoshida K, Yokoi A, Yamamoto T, Hayashi Y, Nakayama J, Yokoi T, Yoshida H, Kato T, Kajiyama H, Yamamoto Y. Aberrant Activation of Cell-Cycle-Related Kinases and the Potential Therapeutic Impact of PLK1 or CHEK1 Inhibition in Uterine Leiomyosarcoma. Clinical cancer research: an official journal of the American Association for Cancer Research, 28:2147-2159, 2022

5. Kawazu M, Ueno T, Saeki K, Sax N, Togashi Y, Kanaseki T, Chida K, Kishigami F, Sato K, Kojima S, Otsuka M, Kawazoe A, Nishinakamura H, Yuka M, Yamamoto Y, Yamashita K, Inoue S, Tanegashima T, Matsubara D, Tane K, Tanaka Y, Iinuma H, Hashiguchi Y, Hazama S, Khor SS, Tokunaga K, Tsuboi M, Niki T, Eto M, Shitara K, Torigoe T, Ishihara S, Aburatani H, Haeno H, Nishikawa H, Mano H. HLA Class I Analysis Provides Insight Into the Genetic and Epigenetic Background of Immune Evasion in Colorectal Cancer With High Microsatellite Instability. Gastroenterology, 162:799-812, 2022

6. Chida K, Kawazoe A, Suzuki T, Kawazu M, Ueno T, Takenouchi K, Nakamura Y, Kuboki Y, Kotani D, Kojima T, Bando H, Mishima S, Kuwata T, Sakamoto N, Watanabe J, Mano H, Ikeda M, Shitara K, Endo I, Nakatsura T, Yoshino T. Transcriptomic Profiling of MSI-H/dMMR Gastrointestinal Tumors to Identify Determinants of Responsiveness to Anti-PD-1 Therapy. Clinical cancer research: an official journal of the American Association for Cancer Research, 28:2110-2117, 2022

7. Yasuda T, Sanada M, Kawazu M, Kojima S, Tsuzuki S, Ueno H, Iwamoto E, Iijima-Yamashita Y, Yamada T, Kanamori T, Nishimura R, Kuwatsuka Y, Takada S, Tanaka M, Ota S, Dobashi N, Yamazaki E, Hirose A, Murayama T, Sumi M, Sato S, Tange N, Nakamura Y, Katsuoka Y, Sakaida E, Kawamata T, Iida H, Shiraishi Y, Nannya Y, Ogawa S, Taniwaki M, Asou N, Hatta Y, Kiyoi H, Matsumura I, Horibe K, Mano H, Naoe T, Miyazaki Y, Hayakawa F. Two novel high-risk adult B-cell acute lymphoblastic leukemia subtypes with high expression of CDX2 and IDH1/2 mutations. Blood, 139:1850-1862, 2022

8. Ando T, Kage H, Shinozaki-Ushiku A, Tatsuno K, Tsutsumi S, Nagayama K, Nakajima J, Kohsaka S, Miyagawa K, Aburatani H, Mano H, Nagase T. Composite Clonal Analysis Reveals Transition of NSCLC Subtypes Through Accumulation of Gene Mutations: A Case Report. JTO clinical and research reports, 3:100277, 2022

9. Ikeuchi H, Hirose T, Ikegami M, Takamochi K, Suzuki K, Mano H, Kohsaka S. Preclinical assessment of combination therapy of EGFR tyrosine kinase inhibitors in a highly heterogeneous tumor model. Oncogene, 41:2470-2479, 2022

10. Nagasaki J, Inozume T, Sax N, Ariyasu R, Ishikawa M, Yamashita K, Kawazu M, Ueno T, Irie T, Tanji E, Morinaga T, Honobe A, Ohnuma T, Yoshino M, Iwata T, Kawase K, Sasaki K, Hanazawa T, Kochin V, Kawamura T, Matsue H, Hino M, Mano H, Suzuki Y, Nishikawa H, Togashi Y. PD-1 blockade therapy promotes infiltration of tumor-attacking exhausted T cell clonotypes. Cell reports, 38:110331, 2022

11. Taguchi A, Rokutan H, Oda K, Tanikawa M, Tanimoto S, Sone K, Mori M, Tsuruga T, Kohsaka S, Tatsuno K, Shinozaki-Ushiku A, Miyagawa K, Mano H, Aburatani H, Ushiku T, Osuga Y. Genetic diagnosis of pseudomyxoma peritonei originating from mucinous borderline tumor inside an ovarian teratoma. BMC medical genomics, 15:51, 2022

12. Kishigami F, Tanaka Y, Yamamoto Y, Ueno T, Kojima S, Sato K, Inoue S, Sugaya S, Ishihara S, Mano H, Kawazu M. Exploration of predictive biomarkers for postoperative recurrence of stage II/III colorectal cancer using genomic sequencing. Cancer medicine, 2022

13. Sasame J, Ikegaya N, Kawazu M, Natsumeda M, Hayashi T, Isoda M, Satomi K, Tomiyama A, Oshima A, Honma H, Miyake Y, Takabayashi K, Nakamura T, Ueno T, Matsushita Y, Iwashita H, Kanemaru Y, Murata H, Ryo A, Terashima K, Yamanaka S, Fujii Y, Mano H, Komori T, Ichimura K, Cahill DP, Wakimoto H, Yamamoto T, Tateishi K. HSP90 Inhibition Overcomes Resistance to Molecular Targeted Therapy in BRAFV600E-mutant High-grade Glioma. Clinical cancer research: an official journal of the American Association for Cancer Research, 28:2425-2439, 2022

14. Odintsov I, Ortiz MV, Khodos I, Mattar MS, Lui AJW, Kohsaka S, de Stanchina E, Bender JLG, Ladanyi M, Somwar R. CIC-Mediated Modulation of MAPK Signaling Opposes Receptor Tyrosine Kinase Inhibitor Response in Kinase-Addicted Sarcoma. Cancer research, 82:1110-1127, 2022

15. Kishikawa S, Hayashi T, Saito T, Takamochi K, Sasa K, Suehara Y, Takahashi F, Sasahara N, Kohsaka S, Suzuki K, Yao T. Distinct properties of pure- and mixed-type high-grade fetal lung adenocarcinomas by genetic profiling and transcription factor expression. Virchows Archiv: an international journal of pathology, 480:609-619, 2022

16. Yamauchi H, Nishimura K, Yoshimi A. Aberrant RNA splicing and therapeutic opportunities in cancers. Cancer science, 113:373-381, 2022

17. Yamaguchi K, Yamamoto T, Chikuda J, Shirota T, Yamamoto Y. Impact of Non-Coding RNAs on Chemotherapeutic Resistance in Oral Cancer. Biomolecules, 12:2022

18. Urabe F, Matsuzaki J, Ito K, Takamori H, Tsuzuki S, Miki J, Kimura T, Egawa S, Nakamura E, Matsui Y, Fujimoto H, Yamamoto Y, Ochiya T. Serum microRNA as liquid biopsy biomarker for the prediction of oncological outcomes in patients with bladder cancer. International journal of urology: official journal of the Japanese Urological Association, 2022

19. Hasegawa N, Kohsaka S, Kurokawa K, Shinno Y, Takeda Nakamura I, Ueno T, Kojima S, Kawazu M, Suehara Y, Ishijima M, Goto Y, Kojima Y, Yonemori K, Hayashi T, Saito T, Shukuya T, Takahashi F, Takahashi K, Mano H. Highly sensitive fusion detection using plasma cell-free RNA in non-small-cell lung cancers. Cancer science, 112:4393-4403, 2021

20. Okuma HS, Yonemori K, Kojima Y, Tanioka M, Sudo K, Noguchi E, Hijioka S, Wakakuwa K, Kato K, Hirakawa A, Kuchiba A, Kubo T, Ichikawa H, Yoshida A, Yatabe Y, Nakamura K, Mano H, Yamamoto N, Fujiwara Y. Clinical Utility of Circulating Tumor DNA in Advanced Rare Cancers. Frontiers in oncology, 11:732525, 2021

21. Kohama I, Asano N, Matsuzaki J, Yamamoto Y, Yamamoto T, Takahashi RU, Kobayashi E, Takizawa S, Sakamoto H, Kato K, Fujimoto H, Chikuda H, Kawai A, Ochiya T. Comprehensive serum and tissue microRNA profiling in dedifferentiated liposarcoma. Oncology letters, 22:623, 2021

22. Ikegami M, Kohsaka S, Hirose T, Ueno T, Inoue S, Kanomata N, Yamauchi H, Mori T, Sekine S, Inamoto Y, Yatabe Y, Kobayashi H, Tanaka S, Mano H. MicroSEC filters sequence errors for formalin-fixed and paraffin-embedded samples. Communications biology, 4:1396, 2021

23. Nakamura IT, Kohsaka S, Ikegami M, Ikeuchi H, Ueno T, Li K, Beyett TS, Koyama T, Shimizu T, Yamamoto N, Takahashi F, Takahashi K, Eck MJ, Mano H. Comprehensive functional evaluation of variants of fibroblast growth factor receptor genes in cancer. NPJ precision oncology, 5:66, 2021

24. Sunami K, Bando H, Yatabe Y, Naito Y, Takahashi H, Tsuchihara K, Toyooka S, Mimori K, Kohsaka S, Uetake H, Kinoshita I, Komine K, Takeda M, Hayashida T, Tamura K, Nishio K, Yamamoto N. Appropriate use of cancer comprehensive genome profiling assay using circulating tumor DNA. Cancer science, 112:3911-3917, 2021

25. Tanaka Y, Chiwaki F, Kojima S, Kawazu M, Komatsu M, Ueno T, Inoue S, Sekine S, Matsusaki K, Matsushita H, Boku N, Kanai Y, Yatabe Y, Sasaki H, Mano H. Multi-omic profiling of peritoneal metastases in gastric cancer identifies molecular subtypes and therapeutic vulnerabilities. Nature cancer, 2:962-977, 2021

26. Chida K, Kawazoe A, Kawazu M, Suzuki T, Nakamura Y, Nakatsura T, Kuwata T, Ueno T, Kuboki Y, Kotani D, Kojima T, Taniguchi H, Mano H, Ikeda M, Shitara K, Endo I, Yoshino T. A Low Tumor Mutational Burden and PTEN Mutations Are Predictors of a Negative Response to PD-1 Blockade in MSI-H/dMMR Gastrointestinal Tumors. Clinical cancer research: an official journal of the American Association for Cancer Research, 27:3714-3724, 2021

27. Inano T, Araki M, Morishita S, Imai M, Kihara Y, Okuda M, Yang Y, Ito M, Osaga S, Mano H, Edahiro Y, Ochiai T, Misawa K, Fukuda Y, Ando J, Komatsu N. Cell-autonomous megakaryopoiesis associated with polyclonal hematopoiesis in triple-negative essential thrombocythemia. Scientific reports, 11:17702, 2021

28. Kawashima S, Inozume T, Kawazu M, Ueno T, Nagasaki J, Tanji E, Honobe A, Ohnuma T, Kawamura T, Umeda Y, Nakamura Y, Kawasaki T, Kiniwa Y, Yamasaki O, Fukushima S, Ikehara Y, Mano H, Suzuki Y, Nishikawa H, Matsue H, Togashi Y. TIGIT/CD155 axis mediates resistance to immunotherapy in patients with melanoma with the inflamed tumor microenvironment. Journal for immunotherapy of cancer, 9:2021

29. Kobayashi H, Makise N, Shinozaki-Ushiku A, Ishibashi Y, Ikegami M, Kohsaka S, Ushiku T, Oda K, Miyagawa K, Aburatani H, Mano H, Tanaka S. Scapular Angiomatoid Fibrous Histiocytoma with EWSR1-CREB1 Fusion in an Adult Patient. Case reports in orthopedics, 2021:9434222, 2021

30. Matsuno Y, Hyodo M, Suzuki M, Tanaka Y, Horikoshi Y, Murakami Y, Torigoe H, Mano H, Tashiro S, Yoshioka KI. Replication-stress-associated DSBs induced by ionizing radiation risk genomic destabilization and associated clonal evolution. iScience, 24:102313, 2021

31. Nagano M, Kohsaka S, Hayashi T, Ueno T, Kojima S, Shinozaki-Ushiku A, Morita S, Tsuda M, Tanaka S, Shinohara T, Omori Y, Sugaya F, Kato H, Narita Y, Nakajima J, Suzuki K, Takamochi K, Mano H. Comprehensive molecular profiling of pulmonary pleomorphic carcinoma. NPJ precision oncology, 5:57, 2021

32. Namba S, Ueno T, Kojima S, Kobayashi K, Kawase K, Tanaka Y, Inoue S, Kishigami F, Kawashima S, Maeda N, Ogawa T, Hazama S, Togashi Y, Ando M, Shiraishi Y, Mano H, Kawazu M. Transcript-targeted analysis reveals isoform alterations and double-hop fusions in breast cancer. Communications biology, 4:1320, 2021

33. Suzuka J, Tsuda M, Wang L, Kohsaka S, Kishida K, Semba S, Sugino H, Aburatani S, Frauenlob M, Kurokawa T, Kojima S, Ueno T, Ohmiya Y, Mano H, Yasuda K, Gong JP, Tanaka S. Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels. Nature biomedical engineering, 5:914-925, 2021

34. Takamochi K, Hara K, Hayashi T, Kohsaka S, Takahashi F, Suehara Y, Suzuki K. Programmed death-ligand 1 expression and its associations with clinicopathological features, prognosis, and driver oncogene alterations in surgically resected lung adenocarcinoma. Lung cancer (Amsterdam, Netherlands), 161:163-170, 2021

35. Sano K, Hayashi T, Suehara Y, Hosoya M, Takamochi K, Kohsaka S, Kishikawa S, Kishi M, Saito S, Takahashi F, Kaneko K, Suzuki K, Yao T, Ishijima M, Saito T. Transcription start site-level expression of thyroid transcription factor 1 isoforms in lung adenocarcinoma and its clinicopathological significance. The journal of pathology. Clinical research, 7:361-374, 2021

36. Yoshioka KI, Kusumoto-Matsuo R, Matsuno Y, Ishiai M. Genomic Instability and Cancer Risk Associated with Erroneous DNA Repair. International journal of molecular sciences, 22:2021

37. De Munck S, Provost M, Kurikawa M, Omori I, Mukohyama J, Felix J, Bloch Y, Abdel-Wahab O, Bazan JF, Yoshimi A, Savvides SN. Structural basis of cytokine-mediated activation of ALK family receptors. Nature, 600:143-147, 2021

38. Miyauchi M, Ito Y, Nakahara F, Hino T, Nakamura F, Iwasaki Y, Kawagoshi T, Koya J, Yoshimi A, Arai S, Kagoya Y, Kurokawa M. Efficient production of human neutrophils from iPSCs that prevent murine lethal infection with immune cell recruitment. Blood, 138:2555-2569, 2021

39. Honda A, Koya J, Yoshimi A, Miyauchi M, Taoka K, Kataoka K, Arai S, Kurokawa M. Loss-of-function mutations in BCOR contribute to chemotherapy resistance in acute myeloid leukemia. Experimental hematology, 101-102:42-48.e11, 2021

40. Maki H, Yoshimi A, Shimada T, Arai S, Morita K, Kamikubo Y, Ikegawa M, Kurokawa M. Physical interaction between BAALC and DBN1 induces chemoresistance in leukemia. Experimental hematology, 94:31-36, 2021

41. Urabe F, Kimura T, Ito K, Yamamoto Y, Tsuzuki S, Miki J, Ochiya T, Egawa S. Urinary extracellular vesicles: a rising star in bladder cancer management. Translational andrology and urology, 10:1878-1889, 2021

42. Yano K, Takahashi RU, Shiotani B, Abe J, Shidooka T, Sudo Y, Yamamoto Y, Kan S, Sakagami H, Tahara H. PRPF19 regulates p53-dependent cellular senescence by modulating alternative splicing of MDM4 mRNA. The Journal of biological chemistry, 297:100882, 2021

43. Kadota T, Fujita Y, Araya J, Watanabe N, Fujimoto S, Kawamoto H, Minagawa S, Hara H, Ohtsuka T, Yamamoto Y, Kuwano K, Ochiya T. Human bronchial epithelial cell-derived extracellular vesicle therapy for pulmonary fibrosis via inhibition of TGF-β-WNT crosstalk. Journal of extracellular vesicles, 10:e12124, 2021

44. Harada K, Sakamoto N, Ukai S, Yamamoto Y, Pham QT, Taniyama D, Honma R, Maruyama R, Takashima T, Ota H, Takemoto Y, Tanabe K, Ohdan H, Yasui W. Establishment of oxaliplatin-resistant gastric cancer organoids: importance of myoferlin in the acquisition of oxaliplatin resistance. Gastric cancer: official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 24:1264-1277, 2021

45. Hashimoto K, Inada M, Yamamoto Y, Ochiya T. Preliminary evaluation of miR-1307-3p in human serum for detection of 13 types of solid cancer using microRNA chip. Heliyon, 7:e07919, 2021

46. Shimomura I, Watanabe N, Yamamoto T, Kumazaki M, Tada Y, Tatsumi K, Ochiya T, Yamamoto Y. Selective targeting of KRAS-driven lung tumorigenesis via unresolved ER stress. JCI insight, 6:2021

47. Yoshida K, Yokoi A, Matsuzaki J, Kato T, Ochiya T, Kajiyama H, Yamamoto Y. Extracellular microRNA profiling for prognostic prediction in patients with high-grade serous ovarian carcinoma. Cancer science, 112:4977-4986, 2021

48. Ueda T, Kanai A, Komuro A, Amano H, Ota K, Honda M, Kawazu M, Okada H. KDM4B promotes acute myeloid leukemia associated with AML1-ETO by regulating chromatin accessibility. FASEB bioAdvances, 3:1020-1033, 2021

49. Nakamura IT, Ikegami M, Hasegawa N, Hayashi T, Ueno T, Kawazu M, Yagishita S, Goto Y, Shinno Y, Kojima Y, Takamochi K, Takahashi F, Takahashi K, Mano H, Kohsaka S. Development of an optimal protocol for molecular profiling of tumor cells in pleural effusions at single-cell level. Cancer science, 112:2006-2019, 2021

50. Suehara Y, Kohsaka S, Hayashi T, Akaike K, Kurisaki-Arakawa A, Sato S, Kobayashi E, Mizuno S, Ueno T, Morii T, Okuma T, Kurihara T, Hasegawa N, Sano K, Sasa K, Okubo T, Kim Y, Mano H, Saito T. Identification of a Novel MAN1A1-ROS1 Fusion Gene Through mRNA-based Screening for Tyrosine Kinase Gene Aberrations in a Patient with Leiomyosarcoma. Clinical orthopaedics and related research, 479:838-852, 2021

51. Yagishita S, Kato K, Takahashi M, Imai T, Yatabe Y, Kuwata T, Suzuki M, Ochiai A, Ohtsu A, Shimada K, Nishida T, Hamada A, Mano H. Characterization of the large-scale Japanese patient-derived xenograft (J-PDX) library. Cancer science, 112:2454-2466, 2021