Regulatory mechanisms of immune checkpoint PD-L1 via the Arf6 pathway in progressive cancer

• Project/Area Number: 17K08614
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Hokkaido University
• Principal Investigator: Hashimoto Ari 北海道大学, 医学研究院, 助教 (60390803)
• Project Period (FY): 2017-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: Arf6 / PD-L1 / 免疫チェクポイント / 免疫回避 / 癌免疫 / 免疫チェックポイント / 免疫監視 / 癌 / シグナル伝達

Outline of Final Research Achievements

We previously showed that the small GTPase Arf6 and its downstream effector AMAP1 constitute the core signaling machinery that drives cancer malignancy and therapeutic resistance in breast and renal cancers. In this project, we showed that mutations in KRAS/TP53, the major driver oncogenes of pancreatic cancer, cooperatively cause the overexpression of Arf6 and AMAP1 proteins and activation of the Arf6-AMAP1 pathway to promote PDAC invasion, metastasis, fibrosis, and immune evasion. Mechanistically, the Arf6-AMAP1 pathway promoted recycling of the immune checkpoint molecule PD-L1 to the cell membrane surface. We also found that the epigenetic factor EZH2 is involved in the regulation of Arf6 and PD-L1 expression, and identified candidate molecules for its regulation. Moreover, we demonstrated that inhibition of the Arf6-AMAP1 pathway in cancer cells results in therapeutic synergy with an anti-PD-1 antibody, and is hence a novel method for improving the efficacy of anti-PD-1 therapies.

Academic Significance and Societal Importance of the Research Achievements

本研究により、膵癌のドライバー変異がArf6-AMAP1経路を高発現活性化することにより、膵癌の悪性度進展を促進し、悪性予後因子となる重要な知見を得た。さらに、エピジェネティック因子EZH2はArf6経路因子及びPD-L1の遺伝子発現に関わること、Arf6-AMAP1経路はPD-L1の細胞内動態制御にも関わり，免疫回避を駆動していることを見出した。実際、Arf6-AMAP1経路阻害は免疫チェックポイント阻害療法に対する抵抗性を改善し、奏功性を相乗的に高めることを実証した。本研究成果により、Arf6-AMAP1の高発現は膵癌の適用バイオマーカーとして有効であり、新規治療戦略の創出が期待出来る。

Research Products

• [Journal Article] Inhibition of mutant KRAS-driven overexpression of ARF6 and MYC by an eIF4A inhibitor drug improves the effects of anti-PD-1 immunotherapy for pancreatic cancer (2021)
  • Author(s): Hashimoto Ari、Handa Haruka、Hata Soichiro、Tsutaho Akio、Yoshida Takao、Hirano Satoshi、Hashimoto Shigeru、Sabe Hisataka
  • Journal Title: Cell Communication and Signaling Volume: 19 Issue: 1 Pages: 54-54
  • DOI: 10.1186/s12964-021-00733-y
  • Peer Reviewed / Open Access
• [Journal Article] Arid5a Promotes Immune Evasion by Augmenting Tryptophan Metabolism and Chemokine Expression (2021)
  • Author(s): Parajuli Gyanu、Tekguc Murat、Wing James B.、Hashimoto Ari、Okuzaki Daisuke、Hirata Takeshi、Sasaki Atsushi、Itokazu Takahide、Handa Haruka、Sugino Hirokazu、Nishikawa Yoshihiro、Metwally Hozaifa、Kodama Yuzo、Tanaka Shinya、Sabe Hisataka、Yamashita Toshihide、Sakaguchi Shimon、Kishimoto Tadamitsu、Hashimoto Shigeru
  • Journal Title: Cancer Immunology Research Volume: 9 Issue: 8 Pages: 862-876
  • DOI: 10.1158/2326-6066.cir-21-0014
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of mutant KRAS-driven overexpression of ARF6 and and MYC by an eIF4A inhibitor drug improves the effects of anti-PD-1 immunotherapy for pancreatic cancer. (2020)
  • Author(s): Hashimoto, A., Handa, H., Hata, S., Tsutaho, A., Yoshida, T., Hirano, S., Hashimoto, S.,and Sabe, H.
  • Journal Title: Cell Communication and Signaling Volume: -
  • Peer Reviewed / Open Access
• [Journal Article] High expression of AMAP1, an ARF6 effector, is associated with elevated levels of PD-L1 and fibrosis of pancreatic cancer. (2020)
  • Author(s): Tsutaho, A., Hashimoto, A., Hashimoto, S., Hata, S., Kachi, S., Hirano, S., and Sabe, H.
  • Journal Title: Cell Communication and Signaling Volume: 18(1) Issue: 1 Pages: 101-101
  • DOI: 10.1186/s12964-020-00608-8
  • Peer Reviewed / Open Access
• [Journal Article] ARF6 and AMAP1 are major targets of KRAS and TP53 mutations to promote invasion, PD-L1 dynamics and immune evasion of pancreatic cancer (2019)
  • Author(s): Hashimoto, S*., Furukawa, S*., Hashimoto, A*., Tsutaho, A., Fukao, A., Sakamura, Y., Parajuli, G., Onodera, Y., Otsuka, Y., Handa, H., Oikawa, T., Hata, S., Nishikawa Y., Mizukami, Y., Kodama, Y., Murakami M., Fujiwara, T., Hirano, S., and Sabe, H. (*Equally contributed)
  • Journal Title: Proceedings of the National Academy of the Sciences of the United States of America Volume: 116 Issue: 35 Pages: 17450-17459
  • DOI: 10.1073/pnas.1901765116
  • Peer Reviewed / Open Access
• [Journal Article] Endothelin type B receptor interacts with the 78-kDa glucose-regulated protein. (2019)
  • Author(s): Mazaki, Y., Higashi, T., Onodera, Y., Nam, JM., Hashimoto, A., Hashimoto, S., Horinouchi, T., and Miwa, S.
  • Journal Title: FEBS Lett Volume: 593(6) Issue: 6 Pages: 644-651
  • DOI: 10.1002/1873-3468.13347
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Epithelial-specific histone modification of the miR-96/182 locus targeting AMAP1 mRNA predisposes p53 to suppress cell invasion in epithelial cells. (2018)
  • Author(s): Handa, H*., Hashimoto, A*., Hashimoto S., Sugino, S., Oikawa, T., and Sabe, H.
  • Journal Title: Cell Commun. Signal. Volume: 16(1) Issue: 1 Pages: 94-94
  • DOI: 10.1186/s12964-018-0302-6
  • NAID: 120006585787
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Frequent overexpression of AMAP1, an Arf6 effector in cell invasion, is characteristic of the MMTV-PyMT rather than the MMTV-Neu human breast cancer model (2018)
  • Author(s): Otsuka Y, Oikawa T, Yoshino H, Hashimoto S, Handa H, Yamamoto H, Hashimoto A, and Sabe H.
  • Journal Title: Cell Commun Signal Volume: 16(1) Issue: 1 Pages: 1-9
  • DOI: 10.1186/s12964-017-0212-z
  • NAID: 120006410532
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prognostic significance of erythrocyte protein band 4.1-like5 expression in upper urinary tract urothelial carcinoma (2017)
  • Author(s): Daimon T, Kosaka T, Kikuchi E, Mikami S, Miyazaki Y, Hashimoto A, Hashimoto S, Mizuno R, Miyajima A, Okada Y, Sabe H, and Oya M
  • Journal Title: Urol Oncol Volume: 35(9) Issue: 9 Pages: 543.e17-543.e24
  • DOI: 10.1016/j.urolonc.2017.04.008
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Blocking the ARF6-AMAP1 pathway cooperatively improves anti-PD-1 immunotherapy for pancreatic cancer (2021)
  • Author(s): Ari Hashimoto, Haruka Handa, Soichiro Hata, Akio Tsutaho, Takao Yoshida, Satoshi Hirano, Shigeru Hashimoto, and Hisataka Sabe
  • Organizer: 第94回日本生化学大会
• [Presentation] Mitochondria after epithelial-mesenchymal transition (EMT) ~ their function and dynamics ~ (2021)
  • Author(s): Haruka Handa, Shingo Takada, Yasuhito Onodera, Yoshizuki Fumoto, Tsukasa Oikawa, Ari Hashimoto, Takashi Yokota, Souichiro Hata, and Hisataka Sabe
  • Organizer: 第94回日本生化学大会
• [Presentation] Pancreatic KRAS/TP53 mutations promote ARF6-based immune evasion via activating mRNA translation and protein prenylation (2021)
  • Author(s): Shigeru Hashimoto, Ari Hashimoto, Shotaro Furukawa, Akio Tsutaho, Yasuhito Onnodera, Yutaro Otsuka, Haruka Handa, Tsukasa Oikawa, Soichiro Hata, Akira Fukao, Yusuke Mizukami, Masaaki Murakami, Toshinobu Fujiwara, Satoshi Hirano, and Hisataka Sabe
  • Organizer: 第7回北大部局横断シンポジウム
• [Presentation] 膵癌におけるARF6-AMAP1経路亢進は浸潤転移制及び免疫抑制的微小環境の誘導に関与する (2020)
  • Author(s): 橋本あり、橋本茂、古川聖太郎、蔦保暁生、半田 悠、畑宗一郎、小野寺康仁、及川司、水上裕輔、平野聡、佐邊壽孝
  • Organizer: 第100回北海道医学大会腫瘍系分科会・第122回北海道癌談話会
• [Presentation] High expression of AMAP1, an ARF6 effector, relates to elevated levels of PD-L1 and fibrosis of pancreatic cancer (2020)
  • Author(s): Ari Hashimoto, Akio Tsutaho, Shigeru Hashimoto, Soichiro Hata, Shion Kachi, Satoshi Hirano, and Hisataka Sabe
  • Organizer: 第79回日本癌学会学術総会
• [Presentation] Upregulation of eIF4A/4E-dependent mRNA translation is the major target of KRAS signaling (2019)
  • Author(s): Hashimoto A., Hashimoto S., Furukawa S., Tsutaho A., Fukao A., Onodera Y., Handa H., Oikawa T., Hata S., Nishikawa Y., Mizukami Y., Kodama Y., Murakami M., Fujiwara T., Hirano S., and Sabe H.
  • Organizer: 第19回蛋白質科学・第71回日本細胞生物合同年次大会（オーガナイザー）
• [Presentation] Pancreatic KRAS/TP53 mutations promote ARF6-based immune evasion via activating mRNA translation and protein prenylation (2019)
  • Author(s): Hashimoto A., Hashimoto S., Furukawa S., Tsutaho A., Fukao A., Onodera Y., Handa H., Oikawa T., Hata S., Nishikawa Y., Mizukami Y., Kodama Y., Murakami M., Fujiwara T., Hirano S., and Sabe H.
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] P53 excludes EZH2 from H3.1 interactome during S phase to maintain histone code (2019)
  • Author(s): Oikawa T., Ohnishi N., Onodera Y., Hashimoto A., Hashimoto A., Ueda K., and Sabe H.
  • Organizer: The 38th Sapporo International Cancer Symposium
  • Int'l Joint Research
• [Presentation] Requirement for p53 in intra-nuclear dynamics of the K27-trimethylated histone H3 during DNA replication (2019)
  • Author(s): Oikawa T., Ohnishi N., Onodera Y., Hashimoto A., Hashimoto A., Ueda K., and Sabe H.
  • Organizer: 第92回日本生化学会
• [Presentation] Pancreatic KRAS and TP53 oncogenes cooperatively activate ARF6-AMAP1 pathway to drive malignancy and immune evasion (2018)
  • Author(s): Ari Hashimoto, Shigeru Hashimoto, Shotaro Furukawa, Akio Tsutaho, Yasuhito Onodera, Yutaro Otsuka, Haruka Handa, Tsukasa Oikawa, Yusuke Mizukami, Masaaki Murakami, Satoshi Hirano, and Hisataka Sabe
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] Requirement for p53 in intra-nuclear dynamics of the K27-trimethylated histone H3 during DNA replication (2018)
  • Author(s): Tsukasa Oikawa, Yuki Shino, Suguru Kurosawa, Yasuhito Onodera, Yutaro Otsuka, Ari Hashimoto, Hisataka Sabe
  • Organizer: Gordon Research Conference 2018 Genomic Instability
• [Presentation] Requirement for p53 in intra-nuclear dynamics of the K27-trimethylated histone H3 during DNA replication (2018)
  • Author(s): Tsukasa Oikawa, Yuki Shino, Suguru Kurosawa, Yasuhito Onodera, Yutaro Otsuka, Ari Hashimoto, Hisataka Sabe
  • Organizer: 第70回日本細胞生物学会（日本発生生物学会合同大会）
• [Presentation] 癌悪性度の代謝側面と癌免疫との関連 (2017)
  • Author(s): 橋本あり
  • Organizer: 第９回シグナルネットワーク研究会
• [Presentation] メバロン酸代謝活性とArf6による癌悪性度進展の分子機序並びにスタチン有効性の解析 (2017)
  • Author(s): 橋本あり、橋本茂、及川司、大塚勇太郎、半田悠、小野寺康仁、佐邊壽孝
  • Organizer: 第116回北海道癌談話会
• [Presentation] 膵癌ドライバー変異はARF6経路を介して癌悪性度とPD-L1発現を促進する (2017)
  • Author(s): 橋本あり、橋本茂、古川聖太郎、蔦保暁生、大塚勇太郎、半田悠、小野寺康仁、及川司、平野聡、佐邊壽孝
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Remarks] http://g21001.med.hokudai.ac.jp/
• [Remarks] 北海道大学大学院医学研究院 生化学分野 分子生物学教室
  • URL: http://g21001.med.hokudai.ac.jp/
• [Patent(Industrial Property Rights)] 癌免疫療法併用剤 (2019)
  • Inventor(s): 佐邊壽孝、平野聡、橋本あり、橋本茂
  • Industrial Property Rights Holder: 佐邊壽孝、平野聡、橋本あり、橋本茂
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
  • Overseas
• [Patent(Industrial Property Rights)] 癌免疫療法併用剤 (2018)
  • Inventor(s): 佐邊壽孝、平野聡、橋本あり、橋本茂
  • Industrial Property Rights Holder: 佐邊壽孝、平野聡、橋本あり、橋本茂
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-048179
  • Filing Date: 2018