Elucidation of the differentiation mechanism of cancer-related fibroblasts using genetically engineered mouse models that develop pancreatic ductal adenocarcinoma

• Project/Area Number: 19K07682
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 50010:Tumor biology-related
• Research Institution: Chiba University
• Principal Investigator: Ikehara Yuzuru 千葉大学, 大学院医学研究院, 教授 (10311440)
• Co-Investigator(Kenkyū-buntansha): 吉富 秀幸 獨協医科大学, 医学部, 教授 (60375631) ; 山口 高志 千葉大学, 大学院医学研究院, 講師 (60626563)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 遺伝子改変膵臓発がんマウスモデル / 間葉系幹細胞 / 癌関連線維芽細胞 / 細胞分化 / がん関連線維芽細胞

Outline of Research at the Start

本研究は、間葉系幹細胞(Mesenchymal Stem Cell [MSC])が、癌関連線維維芽細胞
(Cancer Associated Fibroblast [CAF])の元となり膵管癌間質を構成するようになる分化メカニズムを明らかにし、in vitro三次元培養でその分化過程を再現することである。このため、マウス膵管癌マウスへGFP標識MSC細胞株を移入してCAFを調整して遺伝子発現を解析し、CAFで活性化しているシグナル伝達系路を同定する。さらにGFP標識MSC細胞株に対して、CAFで活性化しているシグナル伝達系路の活性化を誘導することで、分化過程の再現をすすめる。

Outline of Final Research Achievements

We have generated a genetically engineered mouse model (GEM) in which doxycycline administration induces pancreatic ductal adenocarcinoma (PDAC) and dies due to the progression of PDAC about two weeks after administration. Moreover, we revealed that the stromal cells in pancreatic cancer tissue are from bone marrow upon this GEM transplanted with bone marrow from RFP mice.
Furthermore, in in vitro experiments, we recapitulated the CAF differentiation of bone marrow-derived mesenchymal stem cells (MSCs) upon interacting with cancer cells. The gene expression analysis at the single-cell level elucidated the activation of signaling pathways in MSCs towards myofibroblasts differentiation and identified biomarkers detecting CAFs from bone marrow-derived. It is noteworthy that the stroma of human PDAC expressed the identified CAF markers and concluded that we discovered a new path for developing stromal cells in pancreatic ductal adenocarcinoma.

Academic Significance and Societal Importance of the Research Achievements

膵臓癌と診断された患者の平均生存期間は8か月程度であり、これは、化学療法や放射線療法への抵抗性によると考えられている。膵臓癌の治療抵抗性については、膵臓癌組織で観察される豊富ながん間質の出現との関係が指摘されており、治療抵抗性の解消には、間質の発生と進展のメカニズムを理解して対処法を開発することが必要である。本研究は、膵管癌発生の瞬間を再現して、骨髄に由来する細胞が膵臓癌の間質を構成していることを明らかにした。加えて、CAFの同定で利用できるバイオマーカーの開発に成功したことで、膵臓癌の克服につながる研究成果をあげたと考えている。

Research Products

• [Journal Article] Calcification in Werner syndrome associated with lymphatic vessels aging (2021)
  • Author(s): Ogata Hideyuki、Akita Shinsuke、Ikehara Sanae、Azuma Kazuhiko、Yamaguchi Takashi、Maimaiti Maihulan、Maezawa Yoshiro、Kubota Yoshitaka、Yokote Koutaro、Mitsukawa Nobuyuki、Ikehara Yuzuru
  • Journal Title: Aging Volume: 13 Issue: 24 Pages: 25717-25728
  • DOI: 10.18632/aging.203789
  • Peer Reviewed / Open Access
• [Journal Article] The heavy chain of 4F2 antigen promote prostate cancer progression via SKP-2 (2021)
  • Author(s): Maimaiti Maihulan、Sakamoto Shinichi、Sugiura Masahiro、Kanesaka Manato、Fujimoto Ayumi、Matsusaka Keisuke、Xu Minhui、Ando Keisuke、Saito Shinpei、Wakai Ken、Imamura Yusuke、Nakayama Keiichi、Kanai Yoshikatsu、Kaneda Atsushi、Ikehara Yuzuru、Ikeda Jun-Ichiro、Anzai Naohiko、Ichikawa Tomohiko
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 11478-11478
  • DOI: 10.1038/s41598-021-90748-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional analysis of LAT3 in prostate cancer: Its downstream target and relationship with androgen receptor (2021)
  • Author(s): Rii Junryo、Sakamoto Shinichi、Sugiura Masahiro、Kanesaka Manato、Fujimoto Ayumu、Yamada Yasutaka、Maimaiti Maihulan、Ando Keisuke、Wakai Ken、Xu Minhui、Imamura Yusuke、Shindo Norihisa、Hirota Toru、Kaneda Atsushi、Kanai Yoshikatsu、Ikehara Yuzuru、Anzai Naohiko、Ichikawa Tomohiko
  • Journal Title: Cancer Science Volume: 112 Issue: 9 Pages: 3871-3883
  • DOI: 10.1111/cas.14991
  • Peer Reviewed / Open Access
• [Journal Article] TIGIT/CD155 axis mediates resistance to immunotherapy in patients with melanoma with the inflamed tumor microenvironment (2021)
  • Author(s): Kawashima S, Inozume T, Kawazu M, Ueno T, Nagasaki J, Tanji E, Honobe A, Ohnuma T, Kawamura T, Umeda Y, Nakamura Y, Kawasaki T, Kiniwa Y, Yamasaki O, Fukushima S, Ikehara Y, Mano H, Suzuki Y, Nishikawa H, Matsue H, Togashi Y.
  • Journal Title: Journal for ImmunoTherapy of Cancer Volume: 9 Issue: 11 Pages: e003134-e003134
  • DOI: 10.1136/jitc-2021-003134
  • NAID: 120007175403
  • Peer Reviewed / Open Access
• [Journal Article] Brain cell proliferation in adult rats after irradiation with nonequilibrium atmospheric pressure plasma (2021)
  • Author(s): Masanori YAMATO, et al
  • Journal Title: Applied Phisics Express Volume: in press Issue: 6 Pages: 067002-067002
  • DOI: 10.35848/1882-0786/ac03c1
  • Peer Reviewed / Open Access
• [Journal Article] Revision of CHAARTED and LATITUDE criteria among Japanese de novo metastatic prostate cancer patients (2021)
  • Author(s): Kanesaka Manato、Sakamoto Shinichi、Yamada Yasutaka、Rii Junryo、Maimaiti Maihulan、Sazuka Tomokazu、Imamura Yusuke、Komiya Akira、Akakura Koichiro、Ikehara Yuzuru、Nakatsu Hiroomi、Ichikawa Tomohiko
  • Journal Title: Prostate International Volume: 9 Issue: 4 Pages: 208-214
  • DOI: 10.1016/j.prnil.2021.06.001
  • Peer Reviewed / Open Access
• [Journal Article] Effects of electric charges on serum protein aggregation induced by a low temperature atmospheric pressure plasma. (2021)
  • Author(s): Sakakita H, Yamada H, Shimizu T, Fujiwara M, Kato S, Kim J, Ikehara S, Shimizu N, and Ikehara Y.
  • Journal Title: J Phys. D: Appl. Phys. Volume: 54 Pages: 215201-215201
  • Peer Reviewed
• [Journal Article] Detection of Nonpalpable Tiny Axillary Lymph Nodes Surrounded by Adipose Tissue Using a Near-Infrared Camera (2020)
  • Author(s): Akita Shinsuke、Yamaji Yoshihisa、Takeuchi Nobuyoshi、Wakai Ken、Azuma Kazuhiko、Nakagawa Ayako、Fujimoto Hiroshi、Sangai Takafumi、Nagashima Takeshi、Mitsukawa Nobuyuki、Ikehara Yuzuru
  • Journal Title: Lymphatic Research and Biology Volume: 18 Issue: 5 Pages: 455-463
  • DOI: 10.1089/lrb.2019.0022
  • NAID: 130008144970
  • Peer Reviewed
• [Journal Article] Striation phenomena in a low temperature atmospheric pressure neon plasma jet by optical emission spectroscopy (2020)
  • Author(s): Yamada Hiromasa、Kato Susumu、Shimizu Tetsuji、Fujiwara Masanori、Fujiwara Yutaka、Kim Jaeho、Ikehara Sanae、Shimizu Nobuyuki、Ikehara Yuzuru、Sakakita Hajime
  • Journal Title: Physics of Plasmas Volume: 27 Issue: 2 Pages: 022107-022107
  • DOI: 10.1063/1.5124122
  • Peer Reviewed
• [Journal Article] Expression of L-type amino acid transporter 1 as a molecular target for prognostic and therapeutic indicators in bladder carcinoma. (2020)
  • Author(s): Maimaiti M, Sakamoto S, Yamada Y, Sugiura M, Rii J, Takeuchi N, Imamura Y, Furihata T, Ando K, Higuchi K, Xu M, Sazuka T, Nakamura K, Kaneda A, Kanai Y, Kyprianou N, Ikehara Y, Anzai N, Ichikawa T.
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 1292-1292
  • DOI: 10.1038/s41598-020-58136-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mucin-type O-glycosylation controls pluripotency in mouse embryonic stem cells via Wnt receptor endocytosis (2020)
  • Author(s): Pecori Federico、Akimoto Yoshihiro、Hanamatsu Hisatoshi、Furukawa Jun-ichi、Shinohara Yasuro、Ikehara Yuzuru、Nishihara Shoko
  • Journal Title: Journal of Cell Science Volume: 133 Issue: 20 Pages: 245845-245845
  • DOI: 10.1242/jcs.245845
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Dynamics of flow in albumin solution treated by low-temperature atmospheric pressure helium plasma jet (2020)
  • Author(s): Shimizu Tetsuji、Yamada Hiromasa、Fujiwara Masanori、Kato Susumu、Ikehara Yuzuru、Sakakita Hajime
  • Journal Title: AIP Advances Volume: 10 Issue: 12 Pages: 125216-125216
  • DOI: 10.1063/5.0033307
  • Peer Reviewed
• [Journal Article] Cytology-based Detection of Circulating Tumour Cells in Human Pancreatic Cancer Xenograft Models With KRAS Mutation (2020)
  • Author(s): ITO YUKAKO、INOUE ERIKO、MATSUI YUKI、KOBUCHI SHINJI、MOYAMA CHIAMI、AMAGASE KIKUKO、YOSHIMURA MAYUMI、IKEHARA YUZURU、NAKATA SUSUMU、NAKANISHI HAYAO
  • Journal Title: Anticancer Research Volume: 40 Issue: 12 Pages: 6781-6789
  • DOI: 10.21873/anticanres.14701
  • Peer Reviewed
• [Journal Article] TGF-β signaling promotes tube-structure-forming growth in pancreatic duct adenocarcinoma (2019)
  • Author(s): Yamaguchi Takashi、Ikehara Sanae、Akimoto Yoshihiro、Nakanishi Hayao、Kume Masahiko、Yamamoto Kazuo、Ohara Osamu、Ikehara Yuzuru
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 11247-11247
  • DOI: 10.1038/s41598-019-47101-y
  • Peer Reviewed / Open Access
• [Journal Article] A measurement method for determining the correlation between the amount of haemolysis and the electric current in low-temperature plasma treatment. (2019)
  • Author(s): Miyamoto K, Ikehara Y.
  • Journal Title: Plasma Processes and Polymers Volume: 16 Issue: 5 Pages: 1-6
  • DOI: 10.1002/ppap.201800142
  • Peer Reviewed / Open Access
• [Presentation] Development of a new mouse model and imaging sensor technology to elucidate the emergence of pancreatic duct carcinoma (2020)
  • Author(s): Ｉkehara Ｙ, Yamaguchi Ｔ, Ｉkehara S, Hashimoto M, Kusano T, Azuma K
  • Organizer: 第79回日本癌学会学術総会
  • Int'l Joint Research
• [Presentation] プラズマによる帯電調整による血清タンパク質の分散安定性制御 (2019)
  • Author(s): 池原 譲
  • Organizer: 第36回 プラズマ・核融合学会 年会
  • Invited
• [Presentation] Modulation of dispersion stability for serum albumin by through the plasma treatment (2019)
  • Author(s): S. Ikehara, Y. Akimoto, T. Yamaguchi, H. Sakakita, M. Hori, and Y. Ikehara
  • Organizer: 第29回日本MRS年次大会
  • Invited
• [Presentation] Detection of glycosylations by lectins histochemistry, immunohistochemistry and the electron microscopeSymposium：Histochemistry in glycopathology and glycobiology (2019)
  • Author(s): Yuzuru Ikehara, Takashi Yamaguchi, Sanae Ikehara, Kazuhiko Azuma
  • Organizer: 第60回日本組織細胞化学会総会・学術集会/The 13th Japan-China Joint Seminar on Histochemistry and Cytochemistry
  • Int'l Joint Research / Invited
• [Presentation] Low-temperature plasma treatment is a new tissue processing technology - Plasma-induced blood coagulation limits the excessive host responses- (2019)
  • Author(s): Y. Ikehara, S. Ikehara, K. Miyamoto, H. Sakita, K Wakai, N. Takeuchi, T. Yamaguchi
  • Organizer: 24th International Symposium on Plasma Chemistry (ISPC24)
  • Int'l Joint Research / Invited
• [Presentation] 糖鎖情報解析が拓く、Pathogenesis と Disease Progression の理解 (2019)
  • Author(s): 池原 譲
  • Organizer: 第108回日本病理学会総会