Identification of sentization markers and elucidation of drug resistance mechanism for a CK2 inhibitor on cholangiocarcinoma

• Project/Area Number: 19K08449
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Nagoya City University
• Principal Investigator: Miyabe Katsuyuki 名古屋市立大学, 医薬学総合研究院(医学), 研究員 (00543985)
• Co-Investigator(Kenkyū-buntansha): 吉田 道弘 名古屋市立大学, 医薬学総合研究院(医学), 講師 (20636328)
• 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: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 胆管癌 / Silmitasertib / CK2阻害薬 / 薬剤抵抗性 / 胆道癌 / CK2 inhibitor / drug resistance

Outline of Research at the Start

Silmitasertib抵抗性細胞株を作成するとともに、xenograftを使用しRNA sequencingによる薬剤抵抗性遺伝子変異の候補を検出、実際に薬剤抵抗性細胞株から候補遺伝子異常除去後に薬剤感受性が回復するかを検証することで、薬剤抵抗性に関与する遺伝子異常を同定し、関連する薬剤抵抗性因子の存在確認を検体上で行う。次に、胆管癌細胞株に薬剤抵抗性関与遺伝子のクローニングを行うと共にin vitro、in vivoでの薬剤抵抗性の評価も行う。同定された薬剤抵抗性に対する分子標的治療薬追加投与の評価も行い、胆道癌の病理組織標本を用いて、薬剤抵抗性遺伝子発現の臨床検体を用いた確認を行う。

Outline of Final Research Achievements

Exome sequencing was performed on DNA extracted from xenografts obtained from mice treated with Silmitasertib and GEM+CDDP, mice treated with GEM+CDDP, and control mice. Differences such as genetic mutations were investigated, and gene amplification of PGLYRP2 and PCDH86 was also revealed in the comparison between GEM+CDDP and GEM+CDDP+Silmitasertib. Although attempts were made to create drug-resistant strains for in vitro experiments, they have been unsuccessful so far. Therefore, further analysis based on data obtained from in vivo specimens will be continued to elucidate the mechanism of drug resistance.

Academic Significance and Societal Importance of the Research Achievements

CK2阻害剤Silmitasertibの薬剤抵抗性獲得に寄与する機序を検討した。すでにSilmitasertibを投与されたサンプルおよび投与されていないサンプルとの比較をすることにより、薬剤耐性に関わる遺伝子を探り出すことができた。Silmitasertibは近年胆管癌の治療に有望な分子標的薬として注目されており、一部の胆管癌患者の予後を延長するものと期待されている。その薬剤耐性機序を探るうえでのヒントを見つけ出すことにより、本阻害剤が実際の患者に投与され、耐性となった際の次の治療戦略を考える上で、本研究が重要な意義を果たすものと思われる。

Research Products

• [Int'l Joint Research] Mayo Clinic(米国)
• [Int'l Joint Research] メイヨークリニック(米国)
• [Int'l Joint Research] Mayo Clinic(米国)
• [Journal Article] Relevance of gene mutations and methylation to the growth of pancreatic intraductal papillary mucinous neoplasms based on pyrosequencing. (2022)
  • Author(s): Asano, G., Miyabe, K., Kato, H., Yoshida, M., Sawada, T., Okamoto, Y., Sahashi, H., Atsuta, N., Kachi, K., Kato, A., Jinno, N., Natsume, M., Hori, Y., Naitoh, I., Hayashi, K., Matsuo, Y., Takahashi, S., Suzuki, H., Kataoka, H.
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 419-419
  • DOI: 10.1038/s41598-021-04335-z
  • Peer Reviewed / Open Access
• [Journal Article] IgG4-related Sclerosing Cholangitis Complicated with Cholangiocarcinoma and Detected by Forkhead Box P3 Immunohistochemical Staining (2021)
  • Author(s): Toyohara Tadashi、Nakazawa Takahiro、Zakharia Kais、Shimizu Shuya、Miyabe Katsuyuki、Harada Kenichi、Notohara Kenji、Yamada Tomohiro、Hayashi Katsumi、Naitoh Itaru、Hayashi Kazuki、Kataoka Hiromi
  • Journal Title: Internal Medicine Volume: 60 Issue: 6 Pages: 859-866
  • DOI: 10.2169/internalmedicine.5920-20
  • NAID: 130007998464
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2021-03-15
  • Peer Reviewed / Open Access
• [Journal Article] Early Detection of High-grade Biliary Intraepithelial Neoplasia (BilIN-3) in the Cystic Duct Visualized by SpyGlass DS Cholangioscopy (2021)
  • Author(s): Miyabe Katsuyuki、Notohara Kenji、Asano Go、Kato Akihisa、Jinno Naruomi、Natsume Makoto、Hori Yasuki、Yoshida Michihiro、Naitoh Itaru、Tsuboi Ken、Matsuo Yoichi、Ohara Hirotaka、Hayashi Kazuki、Kataoka Hiromi
  • Journal Title: Internal Medicine Volume: 60 Issue: 1 Pages: 47-52
  • DOI: 10.2169/internalmedicine.5072-20
  • NAID: 130007965395
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2021-01-01
• [Journal Article] Over-the-scope-clip treatment for perforation of the duodenum after endoscopic papillectomy (2021)
  • Author(s): Miyabe Katsuyuki、Hori Yasuki、Yoshida Michihiro、Naitoh Itaru、Hayashi Kazuki
  • Journal Title: VideoGIE Volume: 6 Issue: 2 Pages: 101-104
  • DOI: 10.1016/j.vgie.2020.10.001
• [Journal Article] Effect of Statins on the Risk of Extrahepatic Cholangiocarcinoma (2020)
  • Author(s): Lavu Sravanthi、Therneau Terry M.、Harmsen William S.、Mara Kristin C.、Wongjarupong Nicha、Hassan Mohamed、Ali Hamdi A.、Antwi Samuel、Giama Nasra H.、Miyabe Katsuyuki、Roberts Lewis R.
  • Journal Title: Hepatology Volume: - Issue: 4 Pages: 1298-1309
  • DOI: 10.1002/hep.31146
  • Peer Reviewed
• [Journal Article] Novel technique for intraductal cholangioscopy-assisted biliary drainage with over-the-wire microcatheter manipulation. (2019)
  • Author(s): Yoshida M, Kato A, Hayashi K, Naitoh I, Miyabe K, Hori Y, Asano G.
  • Journal Title: Endoscopy Volume: 51 Issue: 12 Pages: 398-399
  • DOI: 10.1055/a-0962-9628
  • Peer Reviewed / Open Access
• [Journal Article] Predictive factors for the mortality of acute pancreatitis on admission. (2019)
  • Author(s): Jinno N, Hori Y, Naitoh I, Miyabe K, Yoshida M, Natsume M, Kato A, Asano G, Sano H, Hayashi K.
  • Journal Title: PLoS One Volume: 14 Issue: 8 Pages: e0221468-e0221468
  • DOI: 10.1371/journal.pone.0221468
  • Peer Reviewed / Open Access
• [Presentation] Relevance of Gene Mutations and Methylation to the Growth of IPMN Based on Pyrosequencing (2022)
  • Author(s): Go Asano
  • Organizer: 第53回日本膵臓学会・第26回国際膵臓学会
  • Int'l Joint Research
• [Presentation] The role of the biliary and gut microbiome in the progression of cholangiocarcinoma and primary sclerosing cholangitis (2019)
  • Author(s): Katsuyuki Miyabe, Vinay Chandrasekhara, Nicha Wongjarupong, Jun Chen, Stephen Johnson, Nicholas Chia, Marina Walther-Antonio, Gregory J. Gores, Mark D. Topazian, Purma C. Kashyap, Lewis R. Roberts, et al.
  • Organizer: American College of Gastoenterology Annual Meeting 2019
  • Int'l Joint Research