高強度ポーラスゲルによる３次元がん組織モデル創出とがん幹細胞標的新規治療法の開発

• Project/Area Number: 21H03802
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: Hokkaido University
• Principal Investigator: 津田 真寿美 北海道大学, 医学研究院, 准教授 (30431307)
• Co-Investigator(Kenkyū-buntansha): 野々山 貴行 北海道大学, 先端生命科学研究院, 准教授 (50709251) ; 田中 伸哉 北海道大学, 医学研究院, 教授 (70261287)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2023: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2022: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2021: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: がん幹細胞 / がん幹細胞ニッシェ / ハイドロゲル / がん治療法 / ポーラスゲル / 癌治療法

Outline of Research at the Start

がんの根絶には、がん組織におけるがん幹細胞の誕生・生存機構の解明が必須である。我々は近年、高強度ハイドロゲルを用いてがん幹細胞を迅速に誘導するリプログラミング現象を見出した（Nature Biomed. Eng. 2021）。これは、高機能ハイドロゲルが分化したがん細胞を未分化ながん幹細胞へ初期化することを可能としたものである。本研究では高強度ポーラスゲルを最適化して3次元がん組織モデルを創出し、がん組織の多種細胞間の時空間的相互作用、およびがん幹細胞が誕生・生存・形質転換する動的分子機構を解明する。最終的にはがん幹細胞を根絶し得る治療標的分子を同定し、がん制圧に向けた基盤の構築を目指す。

Outline of Annual Research Achievements

がんの再発は、がん治療に耐性を示すがん幹細胞が生き残ることによって起きる。癌幹細胞は癌組織中の低酸素や低栄養、および傍血管部位などの特殊な微小環境(癌幹細胞ニッチ)で生成・生存すると考えられているが、その詳細な機序は未だ不明な点が多い。研究代表者らは、近年、高強度ダブルネットワークゲル上にがん細胞を播種すると、24時間以内に迅速にがん幹細胞が誘導されるリプログラミング現象を見出した(Nat. Biomed. Eng., 2021)。当該技術を基盤として、本研究ではがん組織におけるがん幹細胞の誕生・生存機序の解明を目指す。
令和5年度は、極めて予後不良の膠芽腫において、がん細胞ニッチが幹細胞性誘導に与える影響について検討した。ハイドロゲル上でヒト膠芽腫細胞と神経の支持細胞であるアストロサイトを共培養し、両者の直接接触ならびにアストロサイト由来の分泌蛋白が膠芽腫の幹細胞性誘導に与える影響について検討した。共培養ではアストロサイトに接着して膠芽腫細胞が優位に増殖する形態が認められ、検討した４種類のハイドロゲルのうち３種類において幹細胞マーカーのさらなる亢進が認められた。一方、アストロサイト由来の分泌蛋白の影響については、１種類のゲル上で培養したアストロサイトの分泌蛋白が幹細胞性を増加させた。この結果は、脳に存在する支持細胞が至適な環境下で腫瘍の幹細胞性を誘導、およびがん幹細胞の生存に有利に機能していることを示唆する。
さらに、ハイドロゲル上で誘導されたがん幹細胞に対して効果がある薬剤をドラッグスクリーニングした。ヒト膠芽腫細胞を最も幹細胞性の誘導率が高かったハイドロゲル上で培養後に薬剤処理し細胞生存率を検討したところ、複数の膠芽腫細胞株に対して共通して効果が認められた薬剤を複数同定した。これらの薬剤は、高悪性度の膠芽腫に対して生命予後を改善できる可能性があると期待される。

Research Progress Status

令和5年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和5年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] University Hospital Magdeburg(ドイツ)
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• [Journal Article] Geometrical analysis identified morphological features of hydrogel-induced cancer stem cells in synovial sarcoma model cells (2023)
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• [Journal Article] Empagliflozin suppresses mitochondrial reactive oxygen species generation and mitigates the inducibility of atrial fibrillation in diabetic rats (2023)
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  • Journal Title: Frontiers in Cardiovascular Medicine Volume: 10 Pages: 1005408-1005408
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• [Journal Article] Engineering of an electrically charged hydrogel implanted into a traumatic brain injury model for stepwise neuronal tissue reconstruction (2023)
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  • Journal Title: Scientific Reports Volume: 13 Issue: 1 Pages: 2233-2233
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• [Journal Article] Novel rapid immunohistochemistry using an alternating current electric field identifies Rac and Cdc42 activation in human colon cancer FFPE tissues (2022)
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• [Journal Article] Attenuated fusogenicity and pathogenicity of SARS-CoV-2 Omicron variant (2022)
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• [Journal Article] Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike (2022)
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• [Journal Article] Evaluation of the context of downstream N- and free N-glycomic alterations induced by swainsonine in HepG2 cells (2022)
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  • Journal Title: Biochimica et Biophysica Acta (BBA) - General Subjects Volume: 1866 Issue: 9 Pages: 130168-130168
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• [Journal Article] Virological characteristics of the SARS-CoV-2 Omicron BA.2 subvariants, including BA.4 and BA.5 (2022)
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• [Journal Article] Virological characteristics of the SARS-CoV-2 Omicron BA.2.75 variant (2022)
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  • Journal Title: Cell Host & Microbe Volume: 30 Issue: 11 Pages: 1540-1540
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• [Journal Article] Force-triggered rapid microstructure growth on hydrogel surface for on-demand functions (2022)
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  • Journal Title: Nature Communications Volume: 13 Issue: 1 Pages: 6213-6213
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• [Journal Article] Enhanced fusogenicity and pathogenicity of SARS-CoV-2 Delta P681R mutation (2021)
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  • Journal Title: Nature Volume: 602 Issue: 7896 Pages: 300-306
  • DOI: 10.1038/s41586-021-04266-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels (2021)
  • Author(s): Suzuka J, Tsuda M, Wang L, Kohsaka S, Kishida K, Semba S, Sugino H, Aburatani S, Frauenlob M, Kurokawa T, Kojima S, Ueno T, Ohmiya Y, Mano H, Yasuda K, Gong JP, Tanaka S
  • Journal Title: Nat Biomed Eng Volume: － Issue: 8 Pages: 914-925
  • DOI: 10.1038/s41551-021-00692-2
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• [Journal Article] Involvement of BMP and Wnt signals leading to EMT in colon adenocarcinoma with heterotopic ossification. (2021)
  • Author(s): Katono N, Tsuda M, Suzuka J, Oda Y, Wang L, Tanei ZI, Tanino M, Ohata T, Nagabuchi E, Ishida Y, Kimura S, Iwanaga T, Tanaka S.
  • Journal Title: Annals of Clinical and Laboratory Science Volume: 51 Pages: 271-276
  • Peer Reviewed
• [Journal Article] Loss of H3K27 trimethylation is frequent in IDH1-R132H but not in non-canonical IDH1/2 mutated and 1p/19q codeleted oligodendroglioma: a Japanese cohort study (2021)
  • Author(s): Habiba Umma、Sugino Hirokazu、Yordanova Roumyana、Ise Koki、Tanei Zen-ichi、Ishida Yusuke、Tanikawa Satoshi、Terasaka Shunsuke、Sato Ken-ichi、Kamoshima Yuuta、Katoh Masahiko、Nagane Motoo、Shibahara Junji、Tsuda Masumi、Tanaka Shinya
  • Journal Title: Acta Neuropathologica Communications Volume: 9 Issue: 1 Pages: 95-95
  • DOI: 10.1186/s40478-021-01194-7
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive molecular profiling of pulmonary pleomorphic carcinoma (2021)
  • Author(s): Nagano Masaaki、Kohsaka Shinji、Hayashi Takuo、Ueno Toshihide、Kojima Shinya、Shinozaki-Ushiku Aya、Morita Shigeki、Tsuda Masumi、Tanaka Shinya、Shinohara Toshiya、Omori Yuko、Sugaya Fumiko、Kato Hiroaki、Narita Yoshiaki、Nakajima Jun、Suzuki Kenji、Takamochi Kazuya、Mano Hiroyuki
  • Journal Title: npj Precision Oncology Volume: 5 Issue: 1 Pages: 57-57
  • DOI: 10.1038/s41698-021-00201-3
  • Peer Reviewed / Open Access
• [Journal Article] Aberrant expression of MYD88 via RNA‐controlling CNOT4 and EXOSC3 in colonic mucosa impacts generation of colonic cancer (2021)
  • Author(s): Tsuda Masumi、Noguchi Misa、Kurai Tsuyoshi、Ichihashi Yuji、Ise Koki、Wang Lei、Ishida Yusuke、Tanino Mishie、Hirano Satoshi、Asaka Masahiro、Tanaka Shinya
  • Journal Title: Cancer Science Volume: 112 Issue: 12 Pages: 5100-5113
  • DOI: 10.1111/cas.15157
  • Peer Reviewed / Open Access
• [Journal Article] Hydroxyapatite‐hybridized double‐network hydrogel surface enhances differentiation of bone marrow‐derived mesenchymal stem cells to osteogenic cells (2021)
  • Author(s): Kaibara Takuma、Wang Lei、Tsuda Masumi、Nonoyama Takayuki、Kurokawa Takayuki、Iwasaki Norimasa、Gong Jian Ping、Tanaka Shinya、Yasuda Kazunori
  • Journal Title: Journal of Biomedical Materials Research Part A Volume: 110 Issue: 4 Pages: 747-760
  • DOI: 10.1002/jbm.a.37324
  • Peer Reviewed
• [Journal Article] Synthetic poly(2‐acrylamido‐2‐methylpropanesulfonic acid) gel induces chondrogenic differentiation of ATDC5 cells via a novel protein reservoir function (2020)
  • Author(s): Semba Shingo、Kitamura Nobuto、Tsuda Masumi、Goto Keiko、Kurono Sadamu、Ohmiya Yoshihiro、Kurokawa Takayuki、Gong Jian Ping、Yasuda Kazunori、Tanaka Shinya
  • Journal Title: Journal of Biomedical Materials Research Part A Volume: 109 Issue: 3 Pages: 354-364
  • DOI: 10.1002/jbm.a.37028
  • Peer Reviewed / Open Access
• [Presentation] ハイドロゲルが解き明かすがん幹細胞の特性と治療戦略 (2024)
  • Author(s): 津田真寿美
  • Organizer: 第113回日本病理学会総会
• [Presentation] ハイドロゲルを用いた新規治療耐性細胞株の樹立 (2024)
  • Author(s): 王磊、窪田武哲、津田真寿美、小田義崇、平野聡、グン剣萍、田中伸哉
  • Organizer: 第113回日本病理学会総会
• [Presentation] Analysis of stiffness of tumor microenvironment on cancer stemness generation using hydrogels (2024)
  • Author(s): Yanpeng Sun, Yuheng Nie, Masumi Tsuda, Lei Wang, Jian Ping Gong, Shinya Tanaka
  • Organizer: 第113回日本病理学会総会
• [Presentation] Investigation of electric charge on cancer stem cell generation in tumor microenvironment (2024)
  • Author(s): Yuheng Nie, Yanpeng Sun, Masumi Tsuda, Lei Wang, Jian Ping Gong, Shinya Tanaka
  • Organizer: 第113回日本病理学会総会
• [Presentation] TKIs耐性膠芽腫細胞の特性と耐性メカニズムの解析 (2023)
  • Author(s): 津田真寿美、王磊、小田義崇、谷川聖、種井善一、田中伸哉
  • Organizer: 第112回日本病理学会総会
• [Presentation] ハイドロゲルを用いた中皮腫幹細胞の創出および治療標的分子の探索 (2023)
  • Author(s): 加藤万里絵、杉野弘和、津田真寿美、王 磊、種井善一、小田義崇、谷川聖、グン剣萍、田中伸哉
  • Organizer: 第112回日本病理学会総会
• [Presentation] ハイドロゲルを用いた骨肉腫細胞における幹細胞特性の迅速誘導と新規治療標的分子の探索 (2023)
  • Author(s): 甲斐原拓真、王磊、津田真寿美、野々山貴行、黒川孝幸、グン剣萍、岩崎倫政、田中伸哉、安田和則
  • Organizer: 第142回北海道整形災害外科学会
• [Presentation] ハイドロゲルによる幹細胞性誘導を用いて同定したCXCR4の髄膜腫における病理学的、生物学的役割の探索 (2023)
  • Author(s): 小田義崇、津田真寿美、王磊、田中伸哉
  • Organizer: 第27回日本がん分子標的治療学会
• [Presentation] ある種の合成高分子ハイドロゲルは浸潤能の高い滑膜肉腫幹細胞を誘導する (2023)
  • Author(s): 寺島祐樹、津田真寿美、小田義崇、谷川聖、種井善一、王磊、グン剣萍、田中伸哉
  • Organizer: 第40回分子病理学研究会 クラークシンポジウム
• [Presentation] ハイドロゲルを用いた中皮腫幹細胞の創出およびその発現遺伝子の検討 (2023)
  • Author(s): 加藤万里絵、杉野弘和、津田真寿美、王磊、種井 善一、小田義崇、谷川聖、グン剣萍、田中伸哉
  • Organizer: 第40回分子病理学研究会 クラークシンポジウム
• [Presentation] Advancing the Detection of Cancer Stem Cells Using Line Illumination Raman Microscopy and Hydrogel Substrates (2023)
  • Author(s): Jean-Emmanuel Clement, Zannatul Ferdous, Masumi Tsuda, Shinya Tanaka, Jian Ping Gong, Thomas Bocklitz, Katsumasa Fujita, Tamiki Komatsuzaki
  • Organizer: The 11th International Conference on Biological Physics
  • Int'l Joint Research
• [Presentation] Advancing cancer stem cell detection through line illumination Raman microscope and hydrogel substrate (2023)
  • Author(s): Jean-Emmanuel Clement, Zannatul Ferdous, Masumi Tsuda, Shinya Tanaka, Jian Ping Gong, JN Taylor, Thomas Bocklitz, Katsumasa Fujita, Tamiki Komatsuzaki
  • Organizer: The Great SCIentific exchange
  • Int'l Joint Research
• [Presentation] Hydrogel PCDME creates pancreatic cancer stem cells in OXPHOS metabolic state with TXNIP elevation (2023)
  • Author(s): Lei Wang, Yuma Aoki, Masumi Tsuda, Yusuke Saito, Takenori Kubota, Yoshitaka Oda, Satoshi Hirano, Jian Ping Gong, Shinya Tanaka
  • Organizer: The 82nd Annual Meeting of Japanese Cancer Association
• [Presentation] Investigation of cancer stem cell generation by simulating hydrogel- based tumor microenvironment (2023)
  • Author(s): Yuheng Nie, Yanpeng Sun, Masumi Tsuda, Lei Wang, Jian ping Gong, Shinya Tanaka
  • Organizer: The 82nd Annual Meeting of Japanese Cancer Association
• [Presentation] Analysis of the effect of stiffness of cancer microenvironment on stemness induction of cancer cells using hydrogels. (2023)
  • Author(s): Yanpeng Sun, Yuheng Nie, Masumi Tsuda, Lei Wang, Jian ping Gong, Shinya Tanaka
  • Organizer: The 82nd Annual Meeting of Japanese Cancer Association
• [Presentation] Analysis of cancer stem cells at invasive front niche (2022)
  • Author(s): Masumi Tsuda, Shinya Tanaka
  • Organizer: 第81回日本癌学会学術総会
  • Invited
• [Presentation] RNA制御分子CNOT4およびEXOSC3を介したMYD88の発現亢進は大腸癌の発生母地となる (2022)
  • Author(s): 津田真寿美、倉井毅、市橋裕司、伊勢昂生、王磊、石田雄介、谷野美智枝、平野聡、浅香正博、田中伸哉
  • Organizer: 第111回日本病理学会総会
• [Presentation] 肺癌における新規作用機序チロシンキナーゼ阻害剤の解析 (2022)
  • Author(s): 津田真寿美、王磊、田中伸哉
  • Organizer: 第26回日本がん分子標的治療学会学術集会
• [Presentation] Identification of novel LCSCs marker CMKLR1 via reprogramming by Double-Network hydrogel (2022)
  • Author(s): Masumi Tsuda, Michio Tani, Jun Suzuka, Lei Wang, Akinobu Taketomi, Shinya Tanaka
  • Organizer: 第81回日本癌学会学術総会
• [Presentation] Research development and medical application of cancer stem cells using high-functioning hydrogel (2022)
  • Author(s): Masumi Tsuda
  • Organizer: Understanding of novel biological phenomena in cancer and immunology research
  • Int'l Joint Research / Invited
• [Presentation] Analysis of cancer stem cells with invasive capacity by Hydrogel-Activated Reprogramming (HARP) phenomenon using PNaSS hydrogel (2022)
  • Author(s): Masumi Tsuda, Jun Suzuka, Yoshitaka Oda, Lei Wang, Yusuke Saito, Jian Ping Gong, Shinya Tanaka
  • Organizer: 12th AACR-JCA Joint Conference: Breakthroughs in Cancer Research: Translating Knowledge into Practice
  • Int'l Joint Research
• [Presentation] 高分子ハイドロゲルによるリプログラミングを利用した癌幹細胞標的治療薬の同定 (2021)
  • Author(s): 津田真寿美、鈴鹿淳、王磊、田中伸哉
  • Organizer: 第25回日本がん分子標的治療学会学術集会
• [Presentation] 大腸正常粘膜においてRNA制御分子CNOT4およびEXOSC3を介したMYD88の発現亢進は大腸癌の発生に関与する (2021)
  • Author(s): 津田真寿美, 王磊, 平野聡, 浅香正博, 田中伸哉
  • Organizer: 第80回日本癌学会学術総会
• [Patent(Industrial Property Rights)] 細胞培養器、細胞培養キット、および細胞培養方法 (2021)
  • Inventor(s): 黒川孝幸、勝山吉徳、グン剣萍、田中伸哉、津田真寿美
  • Industrial Property Rights Holder: 北海道大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2021-151208
  • Filing Date: 2021