Elucidation of the Mechanisms Governing the Proliferation, Migration and Metastasis of Oral Cancer Cells using Single Cell Analysis

• Project/Area Number: 20H03851
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 57010:Oral biological science-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Watabe Tetsuro 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (00334235)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2022: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2021: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2020: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: 口腔がん / 増殖 / 運動 / 転移 / シングルセル解析 / TGF-β / がん

Outline of Research at the Start

がん細胞は正常細胞が無限の増殖能を獲得することにより生じ、さらに運動能を獲得することで、全身に転移し患者の生命を奪う。がんによる死亡の９割が転移に関連していることを考えると、がんの進展におけるがん細胞の運動能の獲得機構の解明は急務である。本研究においては、腫瘍組織（がん微小環境）における様々な外部因子による口腔がん細胞の増殖と運動の制御がどのように関連しているかという重要な問いを明らかにしたい。

Outline of Final Research Achievements

TGF-β increases epithelial cancer cell migration and metastasis by inducing EMT. TGF-β also inhibits cell proliferation by inducing G1 phase cell-cycle arrest. However, the correlation between these tumor-promoting and -suppressing effects remains unclear. Here, we show that TGF-β confers higher motility and metastatic ability to oral cancer cells in G1 phase. Mechanistically, KRTAP2-3 is a regulator of these dual effects of TGF-β, and its expression is correlated with tumor progression in patients with head and neck cancer and migratory and metastatic potentials of oral cancer cells. Furthermore, scRNA seq reveals that TGF-β generates two populations of mesenchymal cancer cells with differential cell-cycle status through two distinctive EMT pathways mediated by Slug and KRTAP2-3. Thus, TGF-β-induced KRTAP2-3 orchestrates cancer cell proliferation and migration by inducing EMT, suggesting motile cancer cells arrested in G1 phase as a target to suppress metastasis.

Academic Significance and Societal Importance of the Research Achievements

これまで多くの研究者が「増殖をしている細胞が運動する」という通念を持ってきたが、本研究成果により、「増殖をしていない細胞が運動する」という新たな概念を提唱し、基礎細胞生物学におけるパラダイムシフトを起こした点で学術的意義が大きい。また、本研究の成果により新たな診断マーカーならびに治療標的が同定されたため、臨床応用面での波及効果から社会的意義も大きい。

Research Products

• [Journal Article] TGF-β generates a population of cancer cells residing in G1 phase with high motility and metastatic potential via KRTAP2-3 (2022)
  • Author(s): Takahashi Kazuki、Podyma-Inoue Katarzyna A.、Saito Maki、Sakakitani Shintaro、Sugauchi Akinari、Iida Keita、Iwabuchi Sadahiro、Koinuma Daizo、Kurioka Kyoko、Konishi Toru、Tanaka Susumu、Kaida Atsushi、Miura Masahiko、Hashimoto Shinichi、Okada Mariko、Uchihashi Toshihiro、Miyazono Kohei、Watabe Tetsuro
  • Journal Title: Cell Reports Volume: 40 Issue: 13 Pages: 111411-111411
  • DOI: 10.1016/j.celrep.2022.111411
  • Peer Reviewed / Open Access
• [Journal Article] An analysis modality for vascular structures combining tissue-clearing technology and topological data analysis (2022)
  • Author(s): Takahashi K, Abe K, Kubota SI, Fukatsu N, Morishita Y, Yoshimatsu Y, Satoshi Hirakawa, Yoshiaki Kubota, Tetsuro Watabe, Shogo Ehata, Hiroki R. Ueda, Teppei Shimamura, Kohei Miyazono
  • Journal Title: Nature Communiactions Volume: 13 Issue: 1 Pages: 5239-5239
  • DOI: 10.1038/s41467-022-32848-2
  • Peer Reviewed / Open Access
• [Journal Article] Transforming growth factor-β-induced secretion of extracellular vesicles from oral cancer cells evokes endothelial barrier instability via endothelial-mesenchymal transition (2022)
  • Author(s): Kobayashi Miho、Fujiwara Kashio、Takahashi Kazuki、Yoshioka Yusuke、Ochiya Takahiro、Podyma-Inoue Katarzyna A.、Watabe Tetsuro
  • Journal Title: Inflammation and Regeneration Volume: 42 Issue: 1 Pages: 38-38
  • DOI: 10.1186/s41232-022-00225-7
  • Peer Reviewed / Open Access
• [Journal Article] Hapten sensitization to vaginal mucosa induces less recruitment of dendritic cells accompanying TGF‐β‐expressing CD206 ⁺ cells compared with skin (2022)
  • Author(s): Nakayama Kanako、Nishijo Taku、Miyazawa Masaaki、Watabe Tetsuro、Azuma Miyuki、Sakaguchi Hitoshi
  • Journal Title: Immunity, Inflammation and Disease Volume: 10 Issue: 4
  • DOI: 10.1002/iid3.605
  • Peer Reviewed
• [Journal Article] Emerging roles of inflammation-mediated endothelial-mesenchymal transition in health and disease. (2022)
  • Author(s): Yoshimatsu Y, Watabe T.
  • Journal Title: Inflamm Regen Volume: 42 Issue: 1 Pages: 9-9
  • DOI: 10.1186/s41232-021-00186-3
  • Peer Reviewed / Open Access
• [Journal Article] Progression of melanoma is suppressed by targeting all transforming growth factorβ isoforms with an Fc chimeric receptor. (2021)
  • Author(s): Kodama S, Podyma-Inoue KΑ, Uchihashi T, Kurioka K, Takahashi H, Sugauchi A, Takahashi K, Inubushi T, Kogo M, Tanaka S, Watabe T.
  • Journal Title: Oncol Rep Volume: 46 Issue: 3 Pages: 197-197
  • DOI: 10.3892/or.2021.8148
  • Peer Reviewed / Open Access
• [Journal Article] Isolation and characterisation of lymphatic endothelial cells from lung tissues affected by lymphangioleiomyomatosis. (2021)
  • Author(s): Nishino K, Yoshimatsu Y, Muramatsu T, Sekimoto Y, Mitani K, Kobayashi E, Okamoto S, Ebana H, Okada Y, Kurihara M, Suzuki K, Inazawa J, Takahashi K, Watabe T, Seyama K.
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 8406-8406
  • DOI: 10.1038/s41598-021-88064-3
  • Peer Reviewed / Open Access
• [Journal Article] Activation of β2-adrenergic receptor signals suppresses mesenchymal phenotypes of oral squamous cell carcinoma cells. (2021)
  • Author(s): Sakakitani S, Podyma-Inoue KA, Takayama R, Takahashi K, Ishigami-Yuasa M, Kagechika H, Harada H, Watabe T.
  • Journal Title: Cancer Sci Volume: 112 Issue: 1 Pages: 155-167
  • DOI: 10.1111/cas.14670
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Angiogenic Effects of Secreted Factors from Periodontal Ligament Stem Cells (2021)
  • Author(s): Iwasaki Kengo、Akazawa Keiko、Nagata Mizuki、Komaki Motohiro、Peng Yihao、Umeda Makoto、Watabe Tetsuro、Morita Ikuo
  • Journal Title: Dentistry Journal Volume: 9 Issue: 1 Pages: 9-9
  • DOI: 10.3390/dj9010009
  • Peer Reviewed / Open Access
• [Journal Article] ASK1 suppresses NK cell-mediated intravascular tumor cell clearance in lung metastasis. (2021)
  • Author(s): Fujimoto, M., Kamiyama, M., Fuse, K., Ryuno, H., Odawara, T., Furukawa, N., Yoshimatsu, Y., Watabe, T., Michaela, P.-M., Veronika, S., Tahara, H., Hayakawa, Y., Sato, T., Takeda, K., Naguro, I. and Ichijo, H.
  • Journal Title: Cancer Science Volume: 112 Issue: 4 Pages: 1633-1643
  • DOI: 10.1111/cas.14842
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TGF-β and TNF-α cooperatively induce mesenchymal transition of lymphatic endothelial cells via activation of Activin signals (2020)
  • Author(s): Yoshimatsu Y, Kimuro S, Pauty J, Takagaki K, Nomiyama S, Inagawa A, Maeda K, Podyma-InoueKA, Kajiya K, Matsunaga Y, Watabe T
  • Journal Title: PLOS ONE Volume: 15 Issue: 5 Pages: e0232356-e0232356
  • DOI: 10.1371/journal.pone.0232356
  • Peer Reviewed / Open Access
• [Journal Article] TNF-α enhances TGF-β-induced endothelial-to-mesenchymal transition via TGF-β signal augmentation (2020)
  • Author(s): Yasuhiro Yoshimatsu, Ikumi Wakabayashi, Shiori Kimuro, Naoya Takahashi, Kazuki Takahashi, Miho Kobayashi, Nako Maishi, Katarzyna A. Podyma-Inoue, Kyoko Hida, Kohei Miyazono, Tetsuro Watabe
  • Journal Title: Cancer Sciense Volume: - Issue: 7 Pages: 2385-2399
  • DOI: 10.1111/cas.14455
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Peptide-2 from mouse myostatin precursor protein alleviates muscle wasting in cancer-associated cachexia (2020)
  • Author(s): Chiharu Ojima, Yuri Noguchi, Tatsuki Miyamoto, Yuki Saito, Hiroki Orihashi, Yasuhiro Yoshimatsu, Tetsuro Watabe, Kentaro Takayama, Yoshio Hayashi, Fumiko Itoh.
  • Journal Title: Cancer Sci Volume: 111 Issue: 8 Pages: 2954-2964
  • DOI: 10.1111/cas.14520
  • Peer Reviewed / Open Access
• [Journal Article] Targeting all transforming growth factor-β isoforms with an Fc chimeric receptor impairs tumor growth and angiogenesis of oral squamous cell cancer (2020)
  • Author(s): Takahashi Kazuki, Akatsu Yuichi, Podyma-Inoue Katarzyna A., Matsumoto Takehisa, Takahashi Hitomi, Yoshimatsu Yasuhiro, Koinuma Daizo, Shirouzu Mikako, Miyazono Kohei, Watabe Tetsuro
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 36 Pages: 12559-12572
  • DOI: 10.1074/jbc.ra120.012492
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tubulin carboxypeptidase activity of vasohibin-1 inhibits angiogenesis by interfering with endocytosis and trafficking of pro-angiogenic factor receptors (2020)
  • Author(s): Kobayashi Miho、Wakabayashi Ikumi、Suzuki Yasuhiro、Fujiwara Kashio、Nakayama Masanori、Watabe Tetsuro、Sato Yasufumi
  • Journal Title: Angiogenesis Volume: 24 Issue: 1 Pages: 159-176
  • DOI: 10.1007/s10456-020-09754-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Targeting tumor microenvironment networks for developing novel therapeutic strategies (2022)
  • Author(s): Tetsuro Watabe
  • Organizer: Korean Vascular Biology Meeting 2022
  • Int'l Joint Research / Invited
• [Presentation] Roles of TGF-β signals during formation and maintenance of vascular systems (2022)
  • Author(s): Tetsuro Watabe
  • Organizer: 22nd International Vascular Biology Meeting (IVBM2022)
  • Int'l Joint Research / Invited
• [Presentation] Roles of signaling and transcriptional networks during maintenance of vascular systems. (2021)
  • Author(s): Tetsuro Watabe
  • Organizer: 第44回日本分子生物学会
  • Invited
• [Presentation] Roles of TGF-β family signals during progression of oral cancer (2021)
  • Author(s): Tetsuro Watabe
  • Organizer: 13th BMP Conference
  • Int'l Joint Research / Invited
• [Presentation] Activation of epithelial-mesenchymal transition program in oral cancer cells under TGF-β-induced cell cycle arrest. (2021)
  • Author(s): Katarzyna A. Inoue, Kazuki Takahashi, Sakakitani Shintaro, Daizo Koinuma, Akinari Sugauchi, Maki Saito, Atsushi Kaida, Yasuhiro Yoshimatsu, Toshihiro Uchihashi, Masahiko Miura, Kohei Miyazono, Tetsuro Watabe.
  • Organizer: TGF-β Superfamily Conference: Signaling in Development and Disease (FASEB)
  • Int'l Joint Research
• [Presentation] Roles of TGF-β family signals during formation and maintenance of blood and lymphatic vascular systems (2020)
  • Author(s): Tetsuro Watabe
  • Organizer: International Vascular Biology Meeting 2020
  • Int'l Joint Research / Invited
• [Presentation] Oral squamous carcinoma cells under TGF-β-induced cell cycle arrest represent highly motile and invasive population (2020)
  • Author(s): Katarzyna A. Inoue, Kazuki Takahashi, Maki Saito, Atsushi Kaida, Akinari Sugauchi, Toshihiro Uchihashi, Yasuhiro Yoshimatsu, Susumu Tanaka, Masahiko Miura, Mikihiko Kogo, Tetsuro Watabe
  • Organizer: AACR Virtual Special Conference: Tumor Heterogeneity: From Single Cells to Clinical Impact
  • Int'l Joint Research
• [Presentation] TNF-αはTGF-βシグナルの増強を介してTGF-βによって 誘導される血管の内皮間葉移行を促進する (2020)
  • Author(s): 若林育海,吉松康裕,紀室志織,高橋直也,高橋和樹,伊東史子,小林美穂,井上カタジナアンナ,宮園浩平,渡部徹郎
  • Organizer: 第93回日本生化学会大会