Research on Biological and Clinical Significance of VUS in Cancer Clinical Sequencing

2020 Fiscal Year Annual Research Report

• Project/Area Number: 18H02692
• Research Institution: Kyoto University
• Principal Investigator: 武藤 学 京都大学, 医学研究科, 教授 (40360698)
• Co-Investigator(Kenkyū-buntansha): 奥野 恭史 京都大学, 医学研究科, 教授 (20283666) ; 大橋 真也 京都大学, 医学研究科, 特定准教授 (20435556)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 遺伝子変異 / HER2 / APC / がんクリニカルシークエンス

Outline of Annual Research Achievements

がんクリニカルシークエンス検査で検出される、生物学的・臨床的意義の不明な遺伝子変異に関し、当院で解析した大腸癌症例のHER2G776S変異を参考としその生物学的機能解析や標的治療薬の効果を検討した。本研究でHER2G776S変異とAPC変異が相互作用してHER2シグナル伝達に影響することを示した。野生型HER2および変異型HER2-G776Sの発現ベクターを作成し複数のがん細胞株にトランスフェクションすると、HER2G776Sは野生型HER2と比較しHER2シグナルのリン酸化、キナーゼ活性を亢進した。しかしAPC野生型の細胞株（HeLa細胞、FHC細胞）にHER2G776SをトランスフェクションしてもHER2下流シグナル（ERKやAKT）のリン酸化に影響しなかった。一方APC変異のある細胞株（CACO-2、COLO-320）にHER2G776Sのトランスフェクションを行うとERKのリン酸化が増強し足場非依存的増殖能が亢進した。これらよりHER2 G776S変異とAPC変異が相互作用する可能性が示唆された。そこでCRISPR-Cas9法を用いてAPCノックアウトHeLa細胞を樹立し、この細胞にHER2 G776S発現ベクターをトランスフェクションさせると、ERKシグナルの活性化が誘導された。一方APC発現ベクターで野生型APCを高発現させるとHER2-G776S変異によるHER2下流シグナルの活性化は抑制された。これらより変異型HER2 G776S単独でのがん細胞における影響力は低いが、APCの機能喪失を伴うことでがん細胞の悪性能に影響を及ぼす可能性が示唆された。また汎HERチロシンキナーゼ阻害剤であるアファチニブはHER2 G776Sを導入したCOLO-320異種移植片の腫瘍増殖を有意に抑制したことから、このHER2変異は有効な治療標的となると考えられた。

Research Progress Status

令和2年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和2年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Association of local complete response with prognosis after salvage photodynamic therapy for esophageal squamous cell carcinoma. (2021)
  • Author(s): Yusuke Amanuma, Takahiro Horimatsu, Shinya Ohashi, Masashi Tamaoki, Manabu Muto.
  • Journal Title: Dig Endosc. Volume: 33 Pages: 355-363
  • DOI: 10.1111/den.13730.
  • Peer Reviewed / Open Access
• [Journal Article] A nonrandomized, single-arm confirmatory trial of expanded endoscopic submucosal dissection indication for undifferentiated early gastric cancer: Japan Clinical Oncology Group study (JCOG1009/1010) (2021)
  • Author(s): Takizawa K, Ono H, Hasuike N, Takashima A , Minashi K, Boku N , Kushima R , Katayama H, Ogawa G , Fukuda H , Fujisaki J, Oda I, Yano T, Hori S, Doyama H , Hirasawa K, Yamamoto Y, Ishihara R , Tanabe S , Niwa Y, Nakagawa M , Terashima M , Muto M
  • Journal Title: Gastric Cancer. Volume: 24 Pages: 479-491
  • DOI: 10.1007/s10120-020-01134-9
  • Peer Reviewed / Open Access
• [Journal Article] Early gastric cancer detection in high-risk patients: a multicentre randomised controlled trial on the effect of second-generation narrow band imaging. (2021)
  • Author(s): Naohiro Yoshida, Hisashi Doyama, Tomonori Yano, Takahiro Horimatsu, Noriya Uedo, Yoshinobu Yamamoto, Naomi Kakushima, Hiromitsu Kanzaki, Shinichiro Hori, Kenshi Yao, Ichiro Oda, Chikatoshi Katada, Chizu Yokoi, Ken Ohata, Kenichi Yoshimura, Hideki Ishikawa, Manabu Muto.
  • Journal Title: Gut. Volume: 70 Pages: 67-75
  • DOI: 10.1136/gutjnl-2019-319631
  • Peer Reviewed / Open Access
• [Journal Article] Near-focus magnification and second-generation narrow-band imaging for early gastric cancer in a randomized trial (2020)
  • Author(s): Naomi Kakushima, Naohiro Yoshida, Hisashi Doyama, Tomonori Yano, Takahiro Horimatsu, Noriya Uedo, Yoshinobu Yamamoto, Hiromitsu Kanzaki, Shinichiro Hori, Kenshi Yao, Ichiro Oda, Satoshi Tanabe, Chizu Yokoi, Ken Ohata, Kenichi Yoshimura, Hideki Ishikawa & Manabu Muto.
  • Journal Title: J Gastroenterol. Volume: 55 Pages: 1127-1137
  • DOI: 10.1007/s00535-020-01734-3.
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive genomic profiling for patients with chemotherapy-naive advanced cancer. (2020)
  • Author(s): Kondo T, Matsubara J , Pham Nguyen Quy, Fukuyama K, Nomura M, Funakoshi T, Doi K , Sakamori Y, Yoshioka M , Yokoyama A , Tamaoki M, Kou T , Hirohashi K ,Yamada A, Yamamoto Y , Minamiguchi S , Nishigaki M, Yamada T, Kanai M , Matsumoto S , Muto M.
  • Journal Title: Cancer Sci. Volume: 112 Pages: 296-304
  • DOI: 10.1111/cas.14674.
  • Peer Reviewed / Open Access
• [Journal Article] E487K-induced disorder in functionally relevant dynamics of mitochondrial aldehyde dehyderogenase 2. (2020)
  • Author(s): Shigeyuki Matsumoto, Mitsugu Araki, Yuta Isaka, Fumie Ono, Kenshiro Hirohashi, Shinya Ohashi, Manabu Muto, Yasushi Okuno.
  • Journal Title: Biophys J. Volume: 119 Pages: 628-637
  • DOI: 10.1016/j.bpj.2020.07.002.
  • Peer Reviewed / Open Access
• [Journal Article] Cancer of unknown primary with EGFR mutation successfully treated with targeted therapy directed by clinical next-generation sequencing: a case report. (2020)
  • Author(s): Mitani Y, Kanai M, Kou T, Kataoka S, Doi K, Matsubara J, Ohashi S, Matsumoto S, Muto M.
  • Journal Title: BMC Cancer. Volume: 20 Pages: 1177
  • DOI: 10.1186/s12885-020-07640-4.
  • Open Access
• [Journal Article] Protective effects of Alda-1, an ALDH2 activator, on alcohol-derived DNA damage in the esophagus of human ALDH2*2 (Glu504Lys) knock-in mice. (2020)
  • Author(s): Hirohashi K, Ohashi S, Amanuma Y, Nakai Y, Ida T, Baba K, Mitani Y, Mizumoto A, Yamamoto Y, Kikuchi O, Matsubara J, Yamada A, Miyamoto S, Seno H, Matsuda T, Muto M.
  • Journal Title: Carcinogenesis. Volume: 41 Pages: 194-202
  • DOI: 10.1093/carcin/bgz091.
  • Peer Reviewed / Open Access
• [Journal Article] Association between macrocytosis and metachronous squamous cell carcinoma of the esophagus after endoscopic resection in men with early esophageal squamous cell carcinoma. (2020)
  • Author(s): Katada C, Yokoyama T, Yano T, Oda I, Shimizu Y, Doyama H, Koike T, Takizawa K, Hirao M, Okada H, Yoshii T, Kubota Y, Yamanouchi T, Tsuda T, Omori T, Kobayashi N, Suzuki H, Tanabe S, Hori K, Nakayama N, Kawakubo H, Kakushima N, Matsuo Y, Ishikawa H, Yokoyama A, Muto M.
  • Journal Title: Esophagus. Volume: 17 Pages: 149-158
  • DOI: 10.1007/s10388-019-00685-w.
  • Peer Reviewed / Open Access
• [Journal Article] CD24 and CK4 are upregulated by SIM2, and are predictive biomarkers for chemoradiotherapy and surgery in esophageal cancer. (2020)
  • Author(s): Takashima K, Fujii S, Komatsuzaki R, Komatsu M, Takahashi M, Kojima T, Daiko H, Minashi K, Chiwaki F, Muto M, Sasaki H, Yano T.
  • Journal Title: Int J Oncol. Volume: 56 Pages: 835-847
  • DOI: 10.3892/ijo.2020.4963.
  • Peer Reviewed / Open Access
• [Journal Article] The Alcohol Use Disorders Identification Test and the risk of metachronous cancer after endoscopic resection of esophageal cancer. (2020)
  • Author(s): Yokoyama A , Katada C , Yokoyama T , Takizawa K , Yano T, Oda I , Shimizu Y , Nakanishi H , Koike T , Hirao M , Okada H, Yoshii T ,Katagiri A, Yamanouchi T, Matsuo Y , Kawakubo H, Kobayashi N, Ishikawa H, Muto M.
  • Journal Title: Carcinogenesis. Volume: 41 Pages: 1049-1056
  • DOI: 10.1093/carcin/bgaa022.
  • Peer Reviewed / Open Access
• [Journal Article] Synthetic Lethality with Trifluridine/Tipiracil and Checkpoint Kinase 1 Inhibitor for Esophageal Squamous Cell Carcinoma. (2020)
  • Author(s): Shinya Ohashi, Osamu Kikuchi, Yukie Nakai, Tomomi Ida, Tomoki Saito, yuki Kondo, Yoshihiro Yamamoto, Yosuke Mitani, Trang H. Nguyen Vu, Keita Fukuyama, Hiroshi Tsukihara, Norihiko Suzuki, Manabu Muto.
  • Journal Title: Mol Cancer Ther. Volume: 19 Pages: 1363-1372
  • DOI: 10.1158/1535-7163.MCT-19-0918.
  • Peer Reviewed / Open Access
• [Journal Article] Experimental model for the irradiation-mediated abscopal effect and factors influencing this effect. (2020)
  • Author(s): Kiichiro Baba, Motoo Nomura, Shinya Ohashi, Takuya Hiratsuka, Yukie Nakai, Tomoki Saito, Yuki Kondo, Keita Fukuyama, Osamu Kikuchi, Atsushi Yamada, Junichi Matsubara, Kenshiro Hirohashi, Yosuke Mitani, Ayaka Mizumoto, Manabu Muto.
  • Journal Title: Am J Cancer Res. Volume: 10 Pages: 440-453
  • Peer Reviewed / Open Access
• [Journal Article] Clinical significance of TP53 variants as possible secondary findings in tumor-only next-generation sequencing. (2020)
  • Author(s): Yoshihiro Yamamoto, Masashi Kanai, Tadayuki Kou, Aiko Sugiyama, Eijiro Nakamura, Hidehiko Miyake, Takahiro Yamada, Masakazu Nishigaki, Tomohiro Kondo, Hiromi Murakami, Masako Torishima, Shigemi Matsumoto, Shinji Kosugi, and Manabu Muto.
  • Journal Title: J Hum Genet. Volume: 65 Pages: 125-132
  • DOI: 10.1038/s10038-019-0681-6.
  • Peer Reviewed / Open Access
• [Presentation] 食道表在扁平上皮癌内視鏡切除後に発生する異時性頭頸部食道癌とAlcohol Use Disorders Identification Test(AUDIT)の関連 (2020)
  • Author(s): 堅田 親利、横山 顕、武藤 学
  • Organizer: 第62回日本消化器病学会大会
• [Presentation] 食道・大腸の発がんメカニズム (2020)
  • Author(s): 武藤 学
  • Organizer: 第32回広島消化管病態機能研究会
• [Presentation] 頭頚部表在癌全国登録調査－内視鏡診断の検討- (2020)
  • Author(s): 松浦 倫子、鼻岡 昇、堅田 親利、藤井 誠志、横山 徹爾、坂本 泰理、郷田 憲一、石原 立、加納 孝一、門馬 久美子、大森 泰、武藤 学、林 隆一
  • Organizer: 第99回日本消化器内視鏡学会総会
• [Presentation] 本邦における喫煙・飲酒状況の評価を含めた食道内視鏡検査の現状-Japan Endoscopy Database(JED)による検討- (2020)
  • Author(s): 堅田 親利、堀松 高博、武藤 学
  • Organizer: 第99回日本消化器内視鏡学会総会
• [Presentation] 頭頚部表在癌全国登録調査－内視鏡診断の検討 (2020)
  • Author(s): 加納 孝一、鼻岡 昇、堅田 親利、藤井 誠志、郷田 憲一、石原 立、門馬 久美子、大森 泰、武藤 学、林 隆一
  • Organizer: 第44回日本頭頚部癌学会
• [Presentation] 頭頚部・食道における発癌メカニズム (2020)
  • Author(s): 武藤 学、横山 顕礼、大橋 真也
  • Organizer: 第44回日本頭頚部癌学会