Uncovering malignant mechanisms in multiple myeloma cells using CRISPR/Cas9 system

• Project/Area Number: 15K19561
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Hematology
• Research Institution: Aichi Medical University
• Principal Investigator: Ota Akinobu 愛知医科大学, 医学部, 講師 (30438048)
• Research Collaborator: SIVASUNDARAM Karnan
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 多発性骨髄腫 / 悪性化 / Interleukin-6 / cDNAマイクロアレイ / CRISPR/Cas9 / Multiple myeloma / Gene Ontology analysis / GSEA analysis / 細胞増殖 / 遺伝子発現解析 / スクリーニング / ゲノム編集

Outline of Final Research Achievements

Multiple myeloma (MM) is a hematopoietic malignancy with complex and/or different types of genetic background. Although novel therapeutic agents and their combination therapies are more effective than before, MM remains to be an incurable disease. Thus, development of novel molecular-targeted therapies is required to overcome malignant phenotype of MM. Here, the researcher tried to uncover the malignant mechanism of MM using CRISPR/Cas9 system. cDNA microarray analyses revealed that a novel interleukin-6 (IL-6)-inducible serine/threonine kinase X increased after IL-6 treatment in IL-6-dependent MM cell lines, ANBL-6 and FLAM-76. Both mRNA and protein expression of X kinase were detectable and were over-expressed in a subset of MM cell lines. CRISPR/Cas9-mediated gene disruption of gene X resulted in tumor suppression in MM cells. Collectively, these results suggest that novel IL-6-inducible kinase X might be a promising molecular target for MM therapy.

Research Products

• [Journal Article] Delta40p53 suppresses tumor cell proliferation and induces cellular senescence in hepatocellular carcinoma cells (2017)
  • Author(s): Ota A, Nakao H, Sawada Y, Karnan S, Wahiduzzaman M, Inoue T, Kobayashi Y, Yamamoto T, Ishii N, Ohashi T, Nakade Y, Sato K, Itoh K, Konishi H, Hosokawa Y, Yoneda M
  • Journal Title: J Cell Sci Volume: 130 Issue: 3 Pages: 614-625
  • DOI: 10.1242/jcs.190736
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Versican A-subdomain is required for its adequate function in dermal development. (2017)
  • Author(s): Hatano S, Nagai N, Sugiura N, Tsuchimoto J, Isogai z, Kimata K, Ota A, Karnan S, Hosokawa Y, Watanabe H
  • Journal Title: Connect Tissue Res Volume: 印刷中 Issue: 2 Pages: 178-190
  • DOI: 10.1080/03008207.2017.1324432
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Inhibition of Nox1 induces apoptosis by attenuating the AKT signaling pathway in oral squamous cell carcinoma cell lines (2016)
  • Author(s): Ito K, Ota A, Ono T, Nakaoka T, Wahiduzzaman M, Karnan S, Konishi H, Furuhashi A, Hayashi T, Yamada Y, Hosokawa Y, and Kazaoka Y
  • Journal Title: Oncol Rep Volume: 36 Issue: 5 Pages: 2991-2998
  • DOI: 10.3892/or.2016.5068
  • Peer Reviewed
• [Journal Article] Improved methods of AAV-mediated gene targeting for human cell lines using ribosome-skipping 2A peptide (2016)
  • Author(s): Karnan S, Ota A, Konishi Y, Wahiduzzaman M, Hosokawa Y, and Konishi H
  • Journal Title: Nucleic Acids Research Volume: 44 Issue: 6 Pages: e54-e54
  • DOI: 10.1093/nar/gkv1338
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Overexpression of salivary-type amylase reduces the sensitivity to bortezomib in multiple myeloma cells (2015)
  • Author(s): Mizuno, S., Hanamura, I., Ota, A., Karnan, S., Narita, T., Ri, M., Mizutani, M., Goto, M., Gotou, M., Tsunekawa, N., Shikami, M., Iida, S., Hosokawa, Y., Miwa, H., Ueda, R., Nitta, M. and Takami, A
  • Journal Title: Int J Hematol Volume: 102 Issue: 5 Pages: 569-78
  • DOI: 10.1007/s12185-015-1859-0
  • Peer Reviewed / Int'l Joint Research
• [Presentation] NOX1はAKTシグナルを介して口腔扁平上皮がん細胞の生存に寄与する (2016)
  • Author(s): 太田 明伸、カルナン シバスンダラン、小西 裕之、風岡 宜暁、細川 好孝
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2016-10-06
• [Presentation] Combined arsenic trioxide-cisplatin treatment enhances apoptosis in chronic myelogenous leukemia cell lines (2016)
  • Author(s): Md Wahiduzzaman、Akinobu Ota、Sivasundaram Karnan、Hiroyuki Konishi、Yoshitaka Hosokawa
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2016-09-25
• [Presentation] 肝がん細胞株におけるp53アイソフォームdelta40p53の性状解析 (2016)
  • Author(s): 太田 明伸、ワヒドゥザマン ムハマド、カルナン シバスンダラン、小西 裕之、細川 好孝
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2016-09-25
• [Presentation] 悪性胸膜中皮腫の早期診断の腫瘍マーカーの検討 (2016)
  • Author(s): カルナン シバスンダラン、太田 明伸、ワヒドゥーズマン ムハマド、小西 裕之、細川 好孝
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2016-09-25
• [Remarks] 教員紹介 太田 明伸 （おおた あきのぶ）
  • URL: http://www.aichi-med-u.ac.jp/data/profile/Staff_0084.html