Elucidation of the pathogenesis of malignant mesothelioma based on genetic abnormalities and therapeutic targets for genomic medicine

• Project/Area Number: 19K09292
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 55040:Respiratory surgery-related
• Research Institution: Aichi Medical University
• Principal Investigator: Sivasundaram Karnan 愛知医科大学, 医学部, 講師 (30557096)
• Co-Investigator(Kenkyū-buntansha): 村上 秀樹 愛知医科大学, 愛知医科大学, 客員研究員 (90303619)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: MPM / NF2 / p16 / CD24 / ゲノム編集 / 悪性胸膜中皮腫 / p16INK4a / BAP1 / CAMK2D / 脂肪酸合 / デスモソーム / 早期診断マーカー / 新規分子標的薬 / CRISPR-Cas9 / 二重破壊クローン

Outline of Research at the Start

悪性中皮腫は、アスベスト曝露を原因とし発病までに約30-40年を要する。症状確認時の年齢中央値は60歳以後となり特に高齢者に影響している。呼吸困難や息切れなどの自覚症状や画像診断による胸膜の肥厚が認められたときにはすでに進行していることが多い。また、中皮腫ではがん抑制遺伝子である神経線維腫症2型遺伝子NF2、p16, BAP1遺伝子において高頻度に異常が存在しているが中皮腫発症の分子基盤はほとんど明らかになっておらず、早期診断マーカ－や分子標的薬の開発が切望されている。本研究は悪性中皮種の新規標的分子を見出すことが目的である。

Outline of Final Research Achievements

Malignant pleural mesothelioma (MPM) is an aggressive malignancy of the pleura that is currently incurable due to the lack of an effective early diagnostic method and specific medication. The CDKN2A (p16) and NF2 genes are both frequently mutated in MPM. To understand how these mutations contribute to MPM tumor growth, we generated NF2/p16 double-knockout (DKO) cell clones using human MeT-5A and HOMC-B1 mesothelial cell lines. Quantitative PCR analys showed upregulation of CD24 in DKO clones. CD24 was highly expressed in human mesothelioma tissues and associated with the loss of NF2 and p16. Public data analysis revealed a significantly shorter survival time in MPM patients with high CD24 gene expression levels. These results strongly indicate the potential use of CD24 as a prognostic marker as well as a novel diagnostic and therapeutic target for MPM.

Academic Significance and Societal Importance of the Research Achievements

本研究では、NF2欠損に加えてMPM患者で高頻度に見られるp16INK4欠損やBAP1欠損を二重または三重欠損させた細胞を樹立して、その表現型を動物モデルや分子生物学的手法で解明することにより、新たな早期診断マーカーや分子標的治療薬の候補分子の同定が期待できる。本研究から解き明かされる悪性中皮腫の発がん分子病態は極めて高い新規性とゲノム医療への応用が期待できる。また、本研究における解析システムは、他の癌種の分子病態と発がん機構の解析に応用できる可能性を秘めており、その波及的効果は高い。

Research Products

• [Journal Article] Long noncoding RNA profile of the intracranial artery in patients with moyamoya disease (2023)
  • Author(s): Mamiya Takashi、Kanamori Fumiaki、Yokoyama Kinya、Ota Akinobu、Karnan Sivasundaram、Uda Kenji、Araki Yoshio、Maesawa Satoshi、Yoshikawa Kazuhiro、Saito Ryuta
  • Journal Title: Journal of Neurosurgery Volume: 138 Issue: 3 Pages: 709-716
  • DOI: 10.3171/2022.5.jns22579
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Establishment of Mucoepidermoid Carcinoma Cell Lines from Surgical and Recurrence Biopsy Specimens (2023)
  • Author(s): Yamanaka S, Suzuki S, Ito H, Sivasundaram K, Hanamura I, Okubo I, Yoshikawa K, Ono S, Takahara T, Satou A, Tsuzuki T, Ueda R, Ogawa T, Fujimoto Y.
  • Journal Title: International journal of molecular sciences Volume: 24 Issue: 2 Pages: 1722-1722
  • DOI: 10.3390/ijms24021722
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Accumulation of versican and lack of versikine ameliorate acute colitis (2022)
  • Author(s): Islam Shamima、Jahan Nushrat、Shahida Arbee、Karnan Sivasundaram、Watanabe Hideto
  • Journal Title: Matrix Biology Volume: 107 Pages: 59-76
  • DOI: 10.1016/j.matbio.2022.02.004
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cytotoxicity of Callerya speciosa Fractions against Myeloma and Lymphoma Cell Lines (2022)
  • Author(s): Lam Vu Quang、Anh La Hoang、Quan Nguyen Van、Xuan Tran Dang、Hanamura Ichiro、Uchino Kaori、Karnan Sivasundaram、Takami Akiyoshi
  • Journal Title: Molecules Volume: 27 Issue: 7 Pages: 2322-2322
  • DOI: 10.3390/molecules27072322
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Correction of a CD55 mutation to quantify the efficiency of targeted knock-in via flow cytometry (2022)
  • Author(s): Rahman Md. Lutfur、Hyodo Toshinori、Hasan Muhammad Nazmul、Mihara Yuko、Karnan Sivasundaram、Ota Akinobu、Tsuzuki Shinobu、Hosokawa Yoshitaka、Konishi Hiroyuki
  • Journal Title: Molecular Biology Reports Volume: - Issue: 7 Pages: 6241-6248
  • DOI: 10.1007/s11033-022-07422-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell surface expression of human<scp>RP105</scp>depends on<i>N</i>‐glycosylation of<scp>MD</scp>‐1 (2022)
  • Author(s): Biswas Mrityunjoy、Yamazaki Tatsuya、Tomono Susumu、Karnan Sivasundaram、Takagi Hidekazu、Ichimonji Isao、Inui Masanori、Nagaoka Fumiaki、Hosokawa Yoshitaka、Akashi‐Takamura Sachiko
  • Journal Title: FEBS Letters Volume: 596 Issue: 24 Pages: 3211-3231
  • DOI: 10.1002/1873-3468.14452
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BCL6 inhibition ameliorates resistance to ruxolitinib in &lt;i&gt;CRLF2&lt;/i&gt;-rearranged acute lymphoblastic leukemia (2022)
  • Author(s): Tsuzuki Shinobu、Yasuda Takahiko、Goto Hiroaki、Maeda Naoko、Akahane Koshi、Inukai Takeshi、Yamamoto Hideyuki、Karnan Sivasundaram、Ota Akinobu、Hyodo Toshinori、Konishi Hiroyuki、Hosokawa Yoshitaka、Kiyoi Hitoshi、Hayakawa Fumihiko
  • Journal Title: Haematologica Volume: 108 Issue: 2 Pages: 394-408
  • DOI: 10.3324/haematol.2022.280879
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] CD52 is a novel target for the treatment of FLT3-ITD-mutated myeloid leukemia (2021)
  • Author(s): Karnan Sivasundaram、Hanamura Ichiro、Uchino Kaori、Murakami Satsuki、Wahiduzzaman Md、Quang Vu Lam、Rahman Md Lutfur、Hasan Muhammad Nazmul、Hyodo Toshinori、Konishi Hiroyuki、Tsuzuki Shinobu、Yoshikawa Kazuhiro、Suzuki Susumu、Ueda Ryuzo、Ejiri Masayuki、Hosokawa Yoshitaka、Takami Akiyoshi
  • Journal Title: Cell Death Discovery Volume: 7 Issue: 1 Pages: 1-10
  • DOI: 10.1038/s41420-021-00446-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Experimental strategies to achieve efficient targeted knock-in via tandem paired nicking (2021)
  • Author(s): Rahman Md. Lutfur、Hyodo Toshinori、Karnan Sivasundaram、Ota Akinobu、Hasan Muhammad Nazmul、Mihara Yuko、Wahiduzzaman Md、Tsuzuki Shinobu、Hosokawa Yoshitaka、Konishi Hiroyuki
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 1-13
  • DOI: 10.1038/s41598-021-01978-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Flow cytometry-based quantification of targeted knock-in events in human cell lines using a GPI-anchor biosynthesis gene <i>PIGP</i> (2021)
  • Author(s): Rahman Md.?Lutfur、Hyodo Toshinori、Hasan Muhammad?Nazmul、Mihara Yuko、Karnan Sivasundaram、Ota Akinobu、Tsuzuki Shinobu、Hosokawa Yoshitaka、Konishi Hiroyuki
  • Journal Title: Bioscience Reports Volume: 41 Issue: 12
  • DOI: 10.1042/bsr20212231
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chromosomal translocation t(11;14) and p53 deletion induced by the CRISPR/Cas9 system in normal B cell-derived iPS cells (2021)
  • Author(s): Yusuke Azami, Naohiro Tsuyama, Yu Abe, Misaki Sugai-Takahashi, Ken-Ichi Kudo, Akinobu Ota, Karnan Sivasundaram , Moe Muramatsu, Tomonari Shigemura, Megumi Sasatani, Yuko Hashimoto, Shigehira Saji, Kenji Kamiya, Ichiro Hanamura, Takayuki Ikezoe, Masafumi Onodera, Akira Sakai.
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 5216-5216
  • DOI: 10.1038/s41598-021-84628-5
  • Peer Reviewed / Open Access
• [Journal Article] The Clinical and Biological Effects of PD-1 Expression on Tumor Cells in Diffuse Large B-Cell Lymphoma (2021)
  • Author(s): Hanamura Ichiro、Suzuki Susumu、Ota Akinobu、Murakami Satsuki、Satou Akira、Takahara Taishi、Karnan Sivasundaram、Lam Vu Quang、Nakamura Ayano、Takasugi Souichi、Yoshikawa Kazuhiro、Banno Shogo、Ejiri Masayuki、Tsuzuki Toyonori、Hosokawa Yoshitaka、Ueda Ryuzo、Takami Akiyoshi
  • Journal Title: Hemato Volume: 2 Issue: 2 Pages: 368-381
  • DOI: 10.3390/hemato2020023
  • Peer Reviewed / Open Access
• [Journal Article] Transcriptome-wide analysis of intracranial artery in patients with moyamoya disease showing upregulation of immune response, and downregulation of oxidative phosphorylation and DNA repair (2021)
  • Author(s): Kanamori Fumiaki、Yokoyama Kinya、Ota Akinobu、Yoshikawa Kazuhiro、Karnan Sivasundaram、Maruwaka Mikio、Shimizu Kenzo、Ota Shinji、Uda Kenji、Araki Yoshio、Okamoto Sho、Maesawa Satoshi、Wakabayashi Toshihiko、Natsume Atsushi
  • Journal Title: Neurosurgical Focus Volume: 51 Issue: 3 Pages: E3-E3
  • DOI: 10.3171/2021.6.focus20870
  • Peer Reviewed / Open Access
• [Journal Article] Identification of CD24 as a potential diagnostic and therapeutic target for malignant pleural mesothelioma (2020)
  • Author(s): KarnanS, Ota A, Murakami H, Rahman ML, Hasan MN, Wahiduzzaman MD, Hanamura I, Vu LQ, Inoko A, Hyodo T, Konishi H, Tsuzuki S, Hosokawa Y.
  • Journal Title: Cell Death Discovery Volume: 18 Issue: 1 Pages: 127-139
  • DOI: 10.1038/s41420-020-00364-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Discovery of novel molecular characteristics and cellular biological properties in ameloblastoma (2020)
  • Author(s): Kondo Sayuri、Ota Akinobu、Ono Takayuki、Karnan Sivasundaram、Wahiduzzaman Md、Hyodo Toshinori、Lutfur Rahman Md、Ito Kunihiro、Furuhashi Akifumi、Hayashi Tomio、Konishi Hiroyuki、Tsuzuki Shinobu、Hosokawa Yoshitaka、Kazaoka Yoshiaki
  • Journal Title: Cancer Medicine Volume: 9 Issue: 8 Pages: 2904-2917
  • DOI: 10.1002/cam4.2931
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bi-allelic loss of FAM46C triggers tumor growth with concomitant activation of Akt signaling in multiple myeloma cells (2020)
  • Author(s): Kanasugi J, Hanamura I, Ota A, Karnan S, Vu LQ, Mizuno S, Wahiduzzaman M, Rahman ML, Hyodo T, Konishi H, Tsuzuki S, Hosokawa Y, Takami A
  • Journal Title: Cancer Science Volume: - Issue: 5 Pages: 999-999
  • DOI: 10.1111/cas.14386
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Novel Interleukin-6 Inducible Gene PDZ-Binding Kinase Promotes Tumor Growth of Multiple Myeloma Cells (2020)
  • Author(s): Ota Akinobu、Hanamura Ichiro、Karnan Sivasundaram、Inaguma Shingo、Takei Norio、Lam Vu Quang、Mizuno Shohei、Kanasugi Jo、Wahiduzzaman Md、Rahman Md Lutfur、Hyodo Toshinori、Konishi Hiroyuki、Tsuzuki Shinobu、Ikeda Hiroshi、Takami Akiyoshi、Hosokawa Yoshitaka
  • Journal Title: Journal of Interferon & Cytokine Research Volume: 40 Issue: 8 Pages: 389-405
  • DOI: 10.1089/jir.2020.0111
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tandem paired nicking promotes precise genome editing with scarce interference by p53 (2019)
  • Author(s): Hyodo T, Rahman ML, Karnan S, Ito T, Toyoda A, Ota A, Wahiduzzaman M, Tsuzuki S, Okada Y, Hosokawa Y, Konishi H.
  • Journal Title: Cell Rep. Volume: 30 Issue: 4 Pages: 1195-1207
  • DOI: 10.1016/j.celrep.2019.12.064
  • Peer Reviewed / Open Access
• [Presentation] CRISPRCas9を用いたNF2及びCDKN2Aノックアウト人中皮腫細胞株の樹立と分子生物学的解析 (2019)
  • Author(s): 6.Sivasundaram Karnan, Akinobu ota, Ichiro Hanamura, Hideki Murakami, Toshinori Hyodo, Wahiduzzanan Md, Hiroyuki Konishi , Shinobu Tsuzuki, Yoshitaka Hosokawa
  • Organizer: 第92回日本生化学会大会
• [Presentation] P16/NF2遺伝子の欠損により発現の誘導される悪性中皮腫特異的分子の同定 (2019)
  • Author(s): 7.シバスンダラン カルナン、太田明伸、花村一朗、村上秀樹、兵頭寿典、小西裕之、 都築忍、細川好孝
  • Organizer: 第78回日本癌学会学術総会