膵癌間質のプロテオーム解析

2013 Fiscal Year Annual Research Report

• Project/Area Number: 23592018
• Research Institution: Yokohama City University
• Principal Investigator: 遠藤 格 横浜市立大学, 医学(系)研究科(研究院), 教授 (60211091)
• Co-Investigator(Kenkyū-buntansha): 平野 久 横浜市立大学, 生命ナノシステム科学研究科, 教授 (00275075) ; 森 隆太郎 横浜市立大学, 附属病院, 助教 (90596412) ; 廣島 幸彦 横浜市立大学, 医学(系)研究科(研究院), 客員研究員 (60718021)
• Keywords: 膵癌 / 転移 / 浸潤 / ノックアウトマウス / 膵炎 / リン酸化

Research Abstract

【候補蛋白DPYSL3(CRMP4)の機能解析実験】CRMP4が膵癌細胞の浸潤を促進し、静脈浸潤、肝転移を増加させ、膵癌患者の予後を増悪させていることが示唆された（Ann Surg Oncol. 2012.）。
【CRMP4-EGFP, Long formCRMP4-EGFP安定発現株の作成】CRMP4の各isoformの作用を検証するために、CRMP4-EGFP, Long formCRMP4-EGFPのプラスミドを作成し、安定発現株を樹立することを計画した。当該年度においてCRMP4-EGFP, Long formCRMP4-EGFPプラスミドを使用し安定発現株の樹立に成功し、CRMP4の安定発現株で有意にフィロポディア（糸状仮足；細胞の浸潤に不可欠）形成を認めた（未発表データ）。
【CRMP4リン酸化とfilopodiaについて】CRMP4はリン酸化蛋白質であり、リン酸化もより活性化され、その機能を発揮する。サイクリン依存性キナーゼ５(Cdk5)は CRMPのリン酸化に深く関わっていることが知られており、またCdk5の活性化にはp25が必要である。CRMP4高発現膵癌細胞株（CAPAN1）にp25とCDK5をco-transfectionをするとフィロポディアが伸長する傾向を認めた（未発表データ）。
【CRMP4と膵炎】我々はKrasG12D発現マウスとCRMP4ノックアウトマウスを交配させ、KrasG12D発現CRMP4ノックアウトマウスを作成した後に人工的に膵炎を発症させ、CRMP4ノックアウトマウスにおける膵癌モデルを作成する予定であったが、caeruleinによる膵炎惹起方法の検討の中で、CRMP4ノックアウトマウスにおいてより膵炎が重症化する知見を得た（未発表データ）。膵炎の有無でCRMP4自体の発現量を比較したが、差は見られなかった（未発表データ）。これらの結果から、膵炎の重症化にはCRMP4のリン酸化が深く関与している可能性が示唆された。

Research Products

• [Journal Article] Hand-held high-resolution fluorescence imaging system for fluorescence-guided surgery of patient and cell-line pancreatic tumors growing orthotopically in nude mice. (2014)
  • Author(s): Hiroshima Y, Maawy A, Sato S, Murakami T, Uehara F, Miwa S, Yano S, Momiyama M, Chishima T, Tanaka K, Bouvet M, Endo I, Hoffman RM.
  • Journal Title: J Surg Res. Volume: 187(2) Pages: 510-7
  • DOI: 10.1016/j.jss.2013.11.1083
  • Peer Reviewed
• [Journal Article] Efficacy of Salmonella typhimurium A1-R Versus Chemotherapy on a Pancreatic Cancer Patient-Derived Orthotopic Xenograft (PDOX). (2014)
  • Author(s): Hiroshima Y, Zhao M, Maawy A, Zhang Y, Katz MH, Fleming JB, Uehara F, Miwa S, Yano S, Momiyama M, Suetsugu A, Chishima T, Tanaka K, Bouvet M, Endo I, Hoffman RM.
  • Journal Title: J Cell Biochem Volume: 115(7) Pages: 1254-61
  • DOI: 10.1002/jcb.24769
  • Peer Reviewed
• [Journal Article] Successful fluorescence-guided surgery on human colon cancer patient-derived orthotopic xenograft mouse models using a fluorophore-conjugated anti-CEA antibody and a portable imaging system. (2014)
  • Author(s): Hiroshima Y, Maawy A, Metildi CA, Zhang Y, Uehara F, Miwa S, Yano S, Sato S, Murakami T, Momiyama M, Chishima T, Tanaka K, Bouvet M, Endo I, Hoffman RM.
  • Journal Title: J Laparoendosc Adv Surg Tech A. Volume: 24(4) Pages: 241-7
  • DOI: 10.1089/lap.2013.0418
  • Peer Reviewed
• [Journal Article] Collapsin response mediator protein 4 expression is associated with liver metastasis and poor survival in pancreatic cancer. (2013)
  • Author(s): Hiroshima Y, Nakamura F, Miyamoto H, Mori R, Taniguchi K, Matsuyama R, Akiyama H, Tanaka K, Ichikawa Y, Kato S, Kobayashi N, Kubota K, Nagashima Y, Goshima Y, Endo I.
  • Journal Title: Ann Surg Oncol. Volume: 20 Pages: 369-378
  • DOI: 10.1245/s10434-012-2491-3.
  • Peer Reviewed
• [Journal Article] Comparison of efficacy of Salmonella typhimurium A1-R and chemotherapy on stem-like and non-stem human pancreatic cancer cells. (2013)
  • Author(s): Hiroshima Y, Zhao M, Zhang Y, Maawy A, Hassanein MK, Uehara F, Miwa S, Yano S, Momiyama M, Suetsugu A, Chishima T, Tanaka K, Bouvet M, Endo I, Hoffman RM.
  • Journal Title: Cell Cycle Volume: 1;12(17) Pages: 2774-80
  • DOI: 10.4161/cc.25872
  • Peer Reviewed
• [Presentation] Establishment of a patient-derived orthotopic xenograft (PDOX) model of patient cervical cancer (2014)
  • Author(s): Yukihiko Hiroshima1, Yong Zhang1, Ali Maawy2, Sho Sato3, Takashi Murakami3, Mako Yamamoto1, Fuminari Uehara1, Shinji Miwa1, Shuya Yano1, Masashi Momiyama3, Takashi Chishima3, Kuniya Tanaka3, Ali Maawy2, Itaru Endo3, Robert M. Hoffman1.
  • Organizer: AACR
  • Place of Presentation: San Diego, USA
  • Year and Date: 20140405-20140409
• [Presentation] The advantages of patient-derived orthotopic xenograft (PDOX) of metastatic cervical cancer for individualized therapy compared to the PDX model (2014)
  • Author(s): Yukihiko Hiroshima1, Yong Zhang1, Ali Maawy2, Sho Sato3, Takashi Murakami3, Mako Yamamoto1, Fuminari Uehara1, Shinji Miwa1, Shuya Yano1, Masashi Momiyama3, Takashi Chishima3, Kuniya Tanaka3, Michael Bouvet2, Itaru Endo3, Robert M. Hoffman1.
  • Organizer: AACR
  • Place of Presentation: San Diego, USA
  • Year and Date: 20140405-20140409
• [Presentation] Efficacy of Salmonella typhimurium A1-R and anti-VEGF therapy on a patient-derived orthotopic xenograft (PDOX) pancreatic cancer model (2014)
  • Author(s): Yukihiko Hiroshima1, Ming Zhao1, Matthew H.G. Katz2, Jason B. Fleming2, Sho Sato3, Takashi Murakami3, Mako Yamamoto1, Fuminari Uehara1, Shinji Miwa1, Shuya Yano1, Masashi Momiyama3, Yong Zhang1, Ali Maawy4, Takashi Chishima3, Kuniya Tanaka3, Michael Bouvet4, Itaru Endo3, Robert M. Hoffman1.
  • Organizer: AACR
  • Place of Presentation: San Diego, USA
  • Year and Date: 20140405-20140409
• [Presentation] Zoledronic acid inhibits proliferation and metastasis of human pancreatic cancer in the patient-derived orthotopic xenograft (PDOX) model by targeting tumor-educated macrophages (2014)
  • Author(s): Yukihiko Hiroshima1, Mohamed K. Hassenein1, Rhiana Menen1, Matthew H.g. Katz2, Jason B. Fleming2, Sho Sato3, Takashi Murakami3, Mako Yamamoto1, Fuminari Uehara1, Shinji Miwa1, Shuya Yano1, Masashi Momiyama3, Ali Maawy4, Takashi Chishima3, Kuniya Tanaka3, Michael Bouvet4, Itaru Endo3, Robert M. Hoffman1.
  • Organizer: AACR
  • Place of Presentation: San Diego, USA
  • Year and Date: 20140405-20140409
• [Presentation] Efficacy of neoadjuvant chemotherapy in combination with fluorescence-guided surgery on a pancreatic cancer patient-derived orthotopic xenograft (PDOX) (2014)
  • Author(s): Yukihiko Hiroshima1, Ali Maawy1, Yong Zhang1, Sho Sato2, Takashi Murakami2, Mako Yamamoto1, Fuminari Uehara1, Shinji Miwa1, Shuya Yano1, Masashi Momiyama2, Takashi Chishima2, Kuniya Tanaka2, Michael Bouvet3, Itaru Endo2, Robert M. Hoffman1.
  • Organizer: AACR
  • Place of Presentation: San Diego, USA
  • Year and Date: 20140405-20140409
• [Presentation] UVC irradiation in combination with fluorescence-guided surgery cures metastatic human pancreatic cancer in orthotopic mouse models (2014)
  • Author(s): Yukihiko Hiroshima1, Ali Maawy2, Sho Sato3, Takashi Murakami3, Mako Yamamoto1, Fuminari Uehara1, Shinji Miwa1, Shuya Yano1, Masashi Momiyama3, Takashi Chishima3, Kuniya Tanaka3, Michael Bouvet4, Itaru Endo3, Robert M. Hoffman1.
  • Organizer: AACR
  • Place of Presentation: San Diego, USA
  • Year and Date: 20140405-20140409