セツキシマブ長期投与による舌癌細胞の細胞骨格系と細胞内タンパク輸送に及ぼす影響

• Project/Area Number: 21K16942
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 57020:Oral pathobiological science-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 奥山 紘平 東京医科歯科大学, 大学院医歯学総合研究科, 非常勤講師 (30781968)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2023: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 口腔扁平上皮癌 / Gasdermin D / Pyroptosis / Inflammasome / Tumor microenvironment / マクロファージ / EGFR / 頭頸部癌 / 口腔がん / セツキシマブ / ガスダーミンD / パイロトーシス / オートファジー / Hippo-YAP経路 / 4-Nitroquinoline 1-Oxide / Cetuximab / Autophagy / Cell migration / Tubulin / Cytoskeleton / YAP pathway / Cell density / 抗EGFR阻害薬 / 細胞遊走 / 細胞骨格 / 細胞内輸送タンパク / モータータンパク

Outline of Research at the Start

筆者はFucciシステムを導入したSAS細胞(舌癌由来)を用い、セツキシマブ(Cmab)処理後の細胞周期動態を解析し、数日を経てG1期で細胞周期の停止(G1 arrest)を確認し、細胞遊走の有意な低下を認めた。タンパク発現を解析すると、G1 arrestはSkp2の活性低下とp27Kip1の蓄積、オートファジーの発現に起因することが分かった。細胞遊走には細胞骨格系を構成する微小管の動態が関与する。本研究では、Cmab処理によって生じる細胞遊走能の低下と微小管動態の関連、そしてそれによる細胞内輸送タンパクの変化、G1 arrestの生物学的意義を明らかにし、Cmabを用いた治療戦略を探る。

Outline of Annual Research Achievements

セツキシマブを口腔扁平上皮癌細胞株に長期処理をすることによって、細胞遊走能変化や細胞密度上昇が見られたことはすでに論文として報告したが、本研究課題ではさらにそのメカニズムや関連するタンパクの細胞内発現等の変化について詳細を追求した。セツキシマブ長期処理の細胞の運命としてpyroptosis様の細胞死が見られたことから、関連タンパクであるCleaved-Gasdermin D (Gsdmd-N)の発現動態がどのように影響するか、そしてその後の腫瘍組織内の免疫細胞のリクルート、また腫瘍形成への影響を明らかにすることを目標とした。Gsdmdノックアウト(KO)および野生型マウスに4-Nitroquinoline 1-oxide (4NQO)を用い、口腔がん誘発モデルを作成した。口腔内病変形成をエンドポイントとするDisease Free Survivalから、Gsdmd-KOマウスでは有意に口腔粘膜病変形成が遅れ、かつ形成された病変数の減少が見られた。しかし、PCRやウェスタンブロット等により腫瘍の免疫関連遺伝子発現を見ると、有意な変化は見られなかった。これは腫瘍組織の高度な不均一性に由来する結果であると考えられた。一方で、Gsdmdは腫瘍形成において重要な役割を果たすことが示された。
セツキシマブはそのターゲットであるEGFRに働き、下流経路を遮断することで腫瘍の細胞増殖を抑制するのみならず、ADCC活性を有し、抗腫瘍免疫もリクルートする事が知られる。また、細胞内外の様々なタンパクと機能的関連性を有することも報告されている。中でも近年、EGFRとの機能関連タンパクとして、頭頸部扁平上皮癌で高発現する事が報告されているTMEM16Aにおいて、EGFRとその顆粒経路における相互関係、そして細胞の代謝や抗腫瘍免疫における免疫細胞のリクルート等をまとめ、総説として発表した。

Research Products

• [Int'l Joint Research] ミシガン大学/テキサス大学MDアンダーソンがんセンター(米国)
• [Int'l Joint Research] University of Michigan(米国)
• [Int'l Joint Research] University of Michigan(米国)
• [Journal Article] Saturated fatty acids dampen the immunogenicity of cancer by suppressing STING (2023)
  • Author(s): Heath Blake R.、Gong Wang、Taner Hulya F.、Broses Luke、Okuyama Kohei、Cheng Wanqing、Jin Max、Fitzsimonds Zackary R.、Manousidaki Andriana、Wu Yuesong、Zhang Shaoping、Wen Haitao、Chinn Steven B.、Bartee Eric、Xie Yuying、Moon James J.、Lei Yu Leo
  • Journal Title: Cell Reports Volume: 42 Issue: 4 Pages: 112303-112303
  • DOI: 10.1016/j.celrep.2023.112303
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tumor microenvironmental modification by the current target therapy for head and neck squamous cell carcinoma (2023)
  • Author(s): Okuyama Kohei、Naruse Tomofumi、Yanamoto Souichi
  • Journal Title: Journal of Experimental & Clinical Cancer Research Volume: 42 Issue: 1 Pages: 114-114
  • DOI: 10.1186/s13046-023-02691-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell-in-cell structure in cancer: evading strategies from anti-cancer therapies (2023)
  • Author(s): Okuyama Kohei、Fukushima Hiromasa、Naruse Tomofumi、Yanamoto Souichi
  • Journal Title: Frontiers in Oncology Volume: 13 Pages: 1248097-1248097
  • DOI: 10.3389/fonc.2023.1248097
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Oral Bacterial Contributions to Gingival Carcinogenesis and Progression (2023)
  • Author(s): Okuyama Kohei、Yanamoto Souichi
  • Journal Title: Cancer Prevention Research Volume: 16 Issue: 4 Pages: 199-209
  • DOI: 10.1158/1940-6207.capr-22-0511
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Relationship between Tumor Budding and Partial Epithelial?Mesenchymal Transition in Head and Neck Cancer (2023)
  • Author(s): Okuyama Kohei、Suzuki Keiji、Yanamoto Souichi
  • Journal Title: Cancers Volume: 15 Issue: 4 Pages: 1111-1111
  • DOI: 10.3390/cancers15041111
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TMEM16A as a potential treatment target for head and neck cancer (2022)
  • Author(s): Okuyama Kohei、Yanamoto Souichi
  • Journal Title: Journal of Experimental & Clinical Cancer Research Volume: 41 Issue: 1 Pages: 196-196
  • DOI: 10.1186/s13046-022-02405-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical manifestations of diffuse large B-cell lymphoma that exhibits initial symptoms in the maxilla and mandible: a single-center retrospective study (2022)
  • Author(s): Yasuyuki Michi、Hiroyuki Hiroyuki、Yu Oikawa、Kohei Okuyama、Takuma Kugimoto、Takeshi Kuroshima、Hideaki Hirai、Yumi Mochizuki、Hiroaki Shimamoto、Hirofumi Tomioka、Hirokazu Kachi、Jun-ichiro Sakamoto、Kou Kayamori、Tetsuya Yoda
  • Journal Title: BMC Oral Health Volume: 22 Issue: 1 Pages: 20-20
  • DOI: 10.1186/s12903-022-02056-x
  • Peer Reviewed / Open Access
• [Journal Article] Long-term immunosuppressive therapy leads to poor outcomes in patients with oral squamous cell carcinoma. (2022)
  • Author(s): Tomofumi Naruse, Souichi Yanamoto, Mitsunobu Otsuru, Kohei Okuyama, Kota Morishita, Shin-ichi Yamada, Masahiro Umeda
  • Journal Title: Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology Volume: 印刷中
  • Peer Reviewed / Open Access
• [Journal Article] Surgical treatment for oral tongue squamous cell carcinoma: A retrospective study of 432 patients (2021)
  • Author(s): Yu Oikawa, Takuma Kugimoto, Yoshihisa Kashima, Kohei Okuyama, Toshimitsu Ohsako, Takeshi Kuroshima, Hideaki Hirai, Hirofumi Tomioka, Hiroaki Shimamoto, Yasuyuki Michi, Hiroyuki Harada
  • Journal Title: Global Health & Medicine Volume: 3 Issue: 3 Pages: 157-162
  • DOI: 10.35772/ghm.2020.01084
  • NAID: 130008062525
  • ISSN: 2434-9186, 2434-9194
  • Year and Date: 2021-06-30
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Surgical strategy for medication-related osteonecrosis of the jaw (MRONJ) on maxilla: A multicenter retrospective study (2021)
  • Author(s): Kohei Okuyama, Saki Hayashida, Satoshi Rokutanda, Akiko Kawakita, Sakiko Soutome, Shunsuke Sawada, Souichi Yanamoto, Yuka Kojima, Masahiro Umeda
  • Journal Title: Journal of Dental Sciences Volume: - Issue: 3 Pages: 885-890
  • DOI: 10.1016/j.jds.2020.12.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Multicenter retrospective analysis of clinicopathological features and prognosis of oral tongue squamous cell carcinoma in adolescent and young adult patients (2021)
  • Author(s): Kohei Okuyama、Souichi Yanamoto、Yasuyuki Michi、Eri Shibata、Maiko Tsuchiya、Misaki Yokokawa、Tomofumi Naruse、Hirofumi Tomioka、Takeshi Kuroshima、Hiroaki Shimamoto、Tohru Ikeda、Masahiro Umeda、Tetsuya Yoda、Hiroyuki Harada
  • Journal Title: Medicine Volume: 100 Issue: 41 Pages: e27560-e27560
  • DOI: 10.1097/md.0000000000027560
  • Peer Reviewed / Open Access
• [Journal Article] Epithelial-Myoepithelial Carcinoma of the Minor Salivary Glands: Case Series with Comprehensive Review (2021)
  • Author(s): Kohei Okuyama、Yasuyuki Michi、Yoshihisa Kashima、Hirofumi Tomioka、Hideaki Hirai、Misaki Yokokawa、Yuko Yamagata、Takeshi Kuroshima、Yuriko Sato、Maiko Tsuchiya、Kou Kayamoro、Tohru Ikeda、Hiroyuki Harada
  • Journal Title: Diagnostics Volume: 11 Issue: 11 Pages: 2124-2124
  • DOI: 10.3390/diagnostics11112124
  • Peer Reviewed / Open Access
• [Journal Article] Prodromal symptoms of streptococcal toxic shock syndrome caused by Group A Streptococcus in the head and neck region: a case report (2021)
  • Author(s): Kohei Okuyama, Tomofumi Naruse, Souichi Yanamoto, Saki Hayashida, Satoshi Rokutanda, Akiko Kawakita, Kohei Furukawa, MasahiroUmeda
  • Journal Title: Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology Volume: 33 Issue: 2 Pages: 211-214
  • DOI: 10.1016/j.ajoms.2020.08.014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 肺転移に対し外科療法を行った下顎骨原発骨肉腫の1例 (2021)
  • Author(s): 柴田 恵里, 奥山 紘平, 道 泰之, 坂本 啓, 原田 浩之, 依田 哲也
  • Journal Title: 日本口腔外科学会雑誌 Volume: 67 Pages: 353-358
  • NAID: 130008076268
  • Peer Reviewed
• [Presentation] Tumor Microenvironment Modulation by Cetuximab and its Contribution to Subsequent Immune-Checkpoint Inhibition Therapy in Recurrent/Metastatic Head and Neck Squamous Cell Carcinoma (2023)
  • Author(s): Kohei Okuyama, Tomofumi Naruse, Souichi Yamamoto
  • Organizer: 105th American Association of Oral and Maxillofacial Surgeons (AAOMS), Annual Meeting
  • Int'l Joint Research
• [Presentation] BATF2 suppresses cancer initiation by promoting γδ T-cell-mediated immunity (2023)
  • Author(s): Wang Gong, Hulya Taner, Yuesong Wu, Wanqing Cheng, Kohei Okuyama, Zaiye Li, Shadmehr Demehri, Felipe Nör, Deepak Nagrath, Steven B Chinn, Christopher R Donnelly, James J Moon, Yuying Xie, Yu Leo Lei
  • Organizer: IOTN Semi-Annual SC Meeting
  • Int'l Joint Research
• [Presentation] Resolving an Immune Tolerogenic Niche at the Earliest Phase of Oral Cancer Initiation (2023)
  • Author(s): Hulya Taner, Wang Gong, Luke Broses, Kohei Okuyama, Wanqing Cheng, Jung Kuczura, Sashider Rajesh, Yee Sun Tan, Shadmehr Demehri, Jianwen Que, Yuying Xie, Yu Leo Lei
  • Organizer: AACR ANNUAL MEETING 2023
  • Int'l Joint Research
• [Presentation] BATF2 suppresses cancer initiation by promoting γδ T-cell-mediated immunity (2023)
  • Author(s): Wang Gong, Hulya Taner, Yuesong Wu, Wanqing Cheng, Kohei Okuyama, Zaiye Li, Shadmehr Demehri, Felipe Nor, Deepak Nagrath, Steven B Chinn, Christopher R Donnelly, James J Moon, Yuying Xie, Yu Leo Lei
  • Organizer: IOTN Semi-Annual SC Meeting 2023
  • Int'l Joint Research
• [Presentation] IL-1α Mediated Suppressive Myeloid Function in Head and Neck Cancer (2023)
  • Author(s): Hulya F. Taner, Wang Gong, Kohei Okuyama, Luke Proses, Wanqing Cheng, Jung Kuczura, Sashider Rajesh, Yuying Xie, Yu Leo Lei
  • Organizer: The 52nd meeting of the AADOCR, and 47th meeting of the CADR
  • Int'l Joint Research
• [Presentation] Sox2-driven Epithelial Transformation Promotes IL1-mediated Peripheral Immune Tolerance (2023)
  • Author(s): Hulya F. Taner, Wang Gong, Kohei Okuyama, Luke Broses, Wanqing Cheng, Jung Kuczura, Sashider Rajesh, Yuying Xie, Yu Leo Lei
  • Organizer: University of Michigan School of Dentistry Research Day 2023
• [Presentation] TMEM16A as a therapeutic target for Head and Neck Cancer (2022)
  • Author(s): Kohei Okuyama, Souichi Yanamoto
  • Organizer: 8th World Congress of the International Academy of Oral Oncology
  • Int'l Joint Research
• [Presentation] SOX2 Driven Epithelial Transformation Creates Immune Tolerance By Recruiting Suppressive Myeloid Cells (2022)
  • Author(s): Taner H, Broses L, Gong W, Barnett KC, Li Z, Manousidaki A, Nor F, Lanzel E, Tan YS, Okuyama K, Ge S, Zhao H, Polverini PJ, Chinn S, Prince M, Demehri S, Wolf GT, Nor JE, Que J, Ting JP, Xie Y, Lei YL.
  • Organizer: American Association of Dental, Oral, and Craniofacial Research (AADOCR)
  • Int'l Joint Research
• [Presentation] SOX2-driven epithelial transformation creates immune tolerance by recruiting suppressive myeloid cells (2022)
  • Author(s): Taner H, Broses L, Gong W, Barnett KC, Li Z, Manousidaki A, Nor F, Lanzel E, Tan YS, Okuyama K, Ge S, Zhao H, Polverini PJ, Chinn S, Prince M, Demehri S, Wolf GT, Nor JE, Que J, Ting JP, Xie Y, Lei YL.
  • Organizer: University of Michigan School of Dentistry Research Day
  • Int'l Joint Research
• [Presentation] 舌に発生した孔道癌の１例 (2021)
  • Author(s): 時崎 詩織, 富岡 寛文, 伊澤 和倫, 奥山 紘平, 釘本 琢磨, 田中 眞也, 栢森 高, 池田 通, 原田 浩之
  • Organizer: 第211回（公社）日本口腔外科学会関東支部学術集会
• [Book] 月刊「細胞」10月臨時増刊号 特集「がん生物学モデルの最前線」: 頭頸部扁平上皮癌薬物治療が腫瘍微小環境に及ぼす影響 (2023)
  • Author(s): 奥山紘平, 鈴木啓司
  • Total Pages: 5
  • Publisher: 株式会社 ニューサイエンス社