Color-coded intravital imaging demonstrates a transforming growth factor B antagonist selectively targets stromal cells in a human pancreatic-cancer orthotopic mouse model

2016 Fiscal Year Annual Research Report

• Project/Area Number: 26830081
• Research Institution: Yokohama City University
• Principal Investigator: 廣島 幸彦 横浜市立大学, 医学研究科, 客員研究員 (60718021)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 膵癌 / 間質 / SMAD4 / TGFβ / マウスモデル / プロテオーム解析 / realtime imaging

Outline of Annual Research Achievements

膵癌細胞株同所移植モデル、Fluorescent-PDXモデルを作成し、TGFβR1阻害薬（SB-43154）によるTME変化のintravital realtime imagingを行った。その結果、細胞株モデルではTGF-β阻害薬により間質量、微小血管数の減少を認めたが、fluorescent-PDXモデルでは、間質量の減少効果は認められず、むしろ腫瘍量は増大傾向にあった（AACR2016にて発表）。これはOriginalの腫瘍環境をより忠実に再現できるPDXにおいて、細胞株モデルでは殆ど認められなかったTGF-βの抗腫瘍効果が再現されており、それを阻害したために生じたものと考えられ、PDXの有用性を再認識した。
以上の結果から、TGFβR1阻害薬（SB-43154）とゲムシタビンの併用療法の有用性は期待できなかったため、規模を縮小しSMAD4欠失型のPDX同所移植モデルのみを用い、TGFβR1阻害薬とゲムシタビンの併用療法の治療効果について検討した。治療後の腫瘍重量は、①ゲムシタビン単独＜②併用療法＜③コントロール≒④TGFβR1阻害薬単独となり、やはりTGFβR1阻害薬の上乗せ効果は認められなかった。
次に、膵癌の間質増生に関わる新たなターゲット分子を見出すため、SMAD4野生型、欠失型膵癌のTMEにおける発現蛋白の網羅的解析を（LC FT-LTQ MS）を研究協力者平野が施行し、1961種類のタンパク質の同定に成功した。同時にコントロールとして、非癌部間質の解析も同時に施行し、SMAD4欠失型で2倍以上の発現上昇のあった172分子に絞り込んだ。その中から文献的に癌部における細胞外マトリックスの構築に関連していると考えられる以下の3分子を候補分子として選出した。候補分子：①Periostin, ②TenascinC, ③Thrombospondin-1

Research Products

• [Journal Article] Color-coded intravital imaging demonstrates a transforming growth factor-β (TGF-β) antagonist selectively targets stromal cells in a human pancreatic-cancer orthotopic mouse model. (2017)
  • Author(s): Murakami T, Hiroshima Y, Miyake K, Hwang HK, Kiyuna T, DeLong JC, Lwin TM, Matsuyama R, Mori R, Kumamoto T, Chishima T, Tanaka K, Ichikawa Y, Bouvet M, Endo I, Hoffman RM.
  • Journal Title: Cell Cycle Volume: 25 Pages: 1-7
  • DOI: 10.1080/15384101.2017.1315489.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Recombinant methioninase effectively targets a Ewing's sarcoma in a patient-derived orthotopic xenograft (PDOX) nude-mouse model. (2017)
  • Author(s): Murakami T, Li S, Han Q, Tan Y, Kiyuna T, Igarashi K, Kawaguchi K, Hwang HK, Miyake K, Singh AS, Nelson SD, Dry SM, Li Y, Hiroshima Y, Lwin TM, DeLong JC, Chishima T, Tanaka K, Bouvet M, Endo I, Eilber FC, Hoffman RM.
  • Journal Title: Oncotarget Volume: Epub ahead of print Pages: ahead of print
  • DOI: 10.18632/oncotarget.15823.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Patient-derived mouse models of cancer need to be orthotopic in order to evaluate targeted anti-metastatic therapy. (2016)
  • Author(s): Hiroshima Y, Maawy A, Zhang Y, Zhang N, Murakami T, Chishima T, Tanaka K, Ichikawa Y, Bouvet M, Endo I, Hoffman RM.
  • Journal Title: Oncotarget Volume: 7 Pages: 71696-71702
  • DOI: Oncotarget
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Effective fluorescence-guided surgery of liver metastasis using a fluorescent anti-CEA antibody. (2016)
  • Author(s): Hiroshima Y, Lwin TM, Murakami T, Mawy AA, Kuniya T, Chishima T, Endo I, Clary BM, Hoffman RM, Bouvet M.
  • Journal Title: J Surg Oncol Volume: 114 Pages: 951-958
  • DOI: 10.1002/jso.24462.
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Transforming growth factor-beta (TGF-β) inhibitor modulates cancer stroma in a pancreatic cancer orthotopic mouse model (2017)
  • Author(s): Takashi Murakami, Yukihiko Hiroshima, Kentaro Miyake, Tasuku Kiyuna, Ho Kyoung Hwang, Kei Kawaguchi, Jonathan C. DeLong, Thinzer M. Lwin, Kentaro Igarashi, Ryusei Matsuyama, Ryutaro Mori, Takafumi Kumamoto, Takashi Chishima, Kuniya Tanaka, Michael Bouvet, Itaru Endo, Robert M. Hoffman.
  • Organizer: AACR Annual Meeting 2017
  • Place of Presentation: Washington, D.C., USA
  • Year and Date: 2017-04-01 – 2017-04-05
  • Int'l Joint Research
• [Presentation] Tumor-targeting Salmonella typhimurium A1-R promotes CD8+ T cell infiltration in pancreatic cancer in an orthotopic syngeneic mouse model (2016)
  • Author(s): Takashi Murakami, Yukihiko Hiroshima, Yong Zhang, Ming Zhao, Tasuku Kiyuna, Ryusei Matsuyama, Takashi Chishima, Kuniya Tanaka, Michael Bouvet, Itaru Endo, Robert M. Hoffman.
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans, Louisiana, USA
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] Imaging formation of the tumor microenvironment in orthotopic liver tumors in red fluorescent protein (RFP) transgenic nude mice (2016)
  • Author(s): Imaging formation of the tumor microenvironment in orthotopic liver tumors in red fluorescent protein (RFP) transgenic nude mice
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans, Louisiana, USA
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research
• [Presentation] Tumor-targeting Salmonella typhimurium A1-R efficacy on colon cancer liver metastasis alone and in combination with fluorescence-guided surgery (2016)
  • Author(s): Takashi Murakami, Yukihiko Hiroshima, Yong Zhang, Ming Zhao, Ryusei Matsuyama, Takashi Chishima, Kuniya Tanaka, MIchael Bouvet, Itaru Endo, Robert M. Hoffman.
  • Organizer: AACR Annual Meeting 2016
  • Place of Presentation: New Orleans, Louisiana, USA
  • Year and Date: 2016-04-16 – 2016-04-20
  • Int'l Joint Research