Generation and analysis of chronic pancreatitis model mice

• Project/Area Number: 24591016
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Kumamoto University
• Principal Investigator: OHMURAYA Masaki 熊本大学, 生命資源研究・支援センター, 准教授 (60398229)
• Co-Investigator(Renkei-kenkyūsha): ARAKI Kimi 熊本大学, 生命資源研究・支援センター, 教授 (90211705)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 慢性膵炎 / 膵星細胞 / SPINK1/Spink3 / オートファジー / ネクロプトーシス / リポソーム / クロドロネート / Pancreatic stellate cell / Liposome / Chronic pancreatitis

Outline of Final Research Achievements

Mutations in SPINK1 are associated with chronic pancreatitis (CP). Genetic deletion of Spink3, mouse homolog of SPINK1, causes postnatal lethality precluding mechanistic investigations into the effects of SPINK deficiency. Here we have developed SPINK1 knockin in mice by cre-loxP technology (termed “SPINK1-X-in” and “SPINK1- in”). This partial restoration of SPINK function rescues mice from lethality, but, in contrast to control mice, SPINK1-X-in and SPINK1- in mice within 4 weeks developed pancreas damage the earliest manifestations of which were impaired autophagy and increased trypsin activity, and these mice progressively develop spontaneous chronic pancreatitis. SPINK1-in mice represent a novel, clinically relevant, genetic model of human CP, revealing the mechanisms whereby SPINK insufficiency causes CP. The results identify new targets of SPINK regulating autophagic and lysosomal pathways in pancreas.

Research Products

• [Journal Article] Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. (2015)
  • Author(s): Nakagawa, Y., Sakuma, T., Sakamoto, T., Ohmuraya, M., Nakagata, N. and Yamamoto, T.
  • Journal Title: BMC Biotechnol. Volume: -
  • Peer Reviewed / Open Access
• [Journal Article] SPINK1 Status in Colorectal Cancer, Impact on Proliferation, and Role in Colitis-associated Cancer. (2015)
  • Author(s): Ida, S., Ozaki, N., Hirashima, K., Zaitsu, Y., Taki, K., Sakamoto, Y., Miyamoto, Y., Oki, E., Morita, M., Watanabe, M., Maehara, Y., Yamamura, K., Baba, H. and Ohmuraya, M.
  • Journal Title: Mol Cancer Res. Volume: -
  • Peer Reviewed
• [Journal Article] Staging laparoscopy leads to rapid induction of chemotherapy for unresectable pancreatobiliary cancers. (2015)
  • Author(s): Hashimoto, D., Chikamoto, A., Sakata, K., Nakagawa, S., Hayashi, H., Ohmuraya, M., Hirota, M., Yoshida, N., Beppu, T. and Baba, H.
  • Journal Title: Asian journal of endoscopic surgery. Volume: 8 Issue: 1 Pages: 59-62
  • DOI: 10.1111/ases.12138
  • Peer Reviewed / Open Access
• [Journal Article] Clinical features and management of pancreatic solid pseudopapillary tumor. (2014)
  • Author(s): Tsukamoto, M., Hashimoto, D., Chikamoto, A., Abe, S., Ohmuraya, M. and Baba, H.
  • Journal Title: Am Surg. Volume: 80 Pages: 1212-1215
  • Peer Reviewed
• [Journal Article] Generation and analysis of serine protease inhibitor Kazal type 3-Cre driver mice. (2014)
  • Author(s): Sakata, K., Ohmuraya, M., Araki, K., Suzuki, C., Ida, S., Hashimoto, D., Wang, J., Uchiyama, Y., Baba, H. and Yamamura, K.
  • Journal Title: Exp Anim. Volume: 63 Pages: 45-53
  • NAID: 130003391605
  • Peer Reviewed
• [Journal Article] Autophagy regulation in the pancreatic acinar cells is independent of epidermal growth factor receptor signaling. (2014)
  • Author(s): Ozaki, N., Fukuchi, Y., Tomiyoshi, S., Uehara1, H., Ida, S., Wang, J., Kimi Araki, K., Sibilia, M., Baba, H., and Yamamura, K. and Ohmuraya, M.
  • Journal Title: Biochem. Biophysic. Res. Comm. Volume: 446 Issue: 1 Pages: 224-230
  • DOI: 10.1016/j.bbrc.2014.02.111
  • Peer Reviewed / Open Access
• [Journal Article] Application of Oocyte Cryopreservation Technology in TALEN-Mediated Mouse Genome Editing. (2014)
  • Author(s): Nakagawa, Y., Sakuma, T., Nakagata, N., Yamasaki, S., Takeda, N., Ohmuraya, M. and Yamamoto, T.
  • Journal Title: Exp Anim. Volume: 63 Pages: 349-355
  • NAID: 130004677827
  • Peer Reviewed
• [Journal Article] Impact of Postoperative Weight Loss on Survival After Resection for Pancreatic Cancer. (2014)
  • Author(s): Hashimoto, D., Chikamoto, A., Ohmuraya, M., Abe, S., Nakagawa, S., Beppu, T., Takamori, H., Hirota, M. and Baba, H.
  • Journal Title: JPEN J Parenter Enteral Nutr. Volume: -
  • Peer Reviewed
• [Journal Article] Impact of Postoperative Weight Loss on Survival After Resection for Pancreatic Cancer. (2014)
  • Author(s): Hashimoto D, Chikamoto A, Ohmuraya M, Abe S, Nakagawa S, Beppu T, Takamori H, Hirota M, Baba H
  • Journal Title: JPEN J Parenter Enteral Nutr. Volume: -
  • Peer Reviewed
• [Journal Article] Generation and analysis of serine protease inhibitor kazal type 3-cre driver mice. (2014)
  • Author(s): Sakata K, Ohmuraya M, Araki K, Suzuki C, Ida S, Hashimoto D, Wang J, Uchiyama Y, Baba H, Yamamura K
  • Journal Title: Exp Anim. Volume: 63 Pages: 45-53
  • NAID: 130003391605
  • Peer Reviewed
• [Journal Article] Screening methods to identify TALEN-mediated knockout mice. (2014)
  • Author(s): Nakagawa Y, Yamamoto T, Suzuki K, Araki K, Takeda N, Ohmuraya M, Sakuma T
  • Journal Title: Exp Anim. Volume: 63 Pages: 79-84
  • NAID: 130003391606
  • Peer Reviewed
• [Journal Article] Pancreatic cancer in the remnant pancreas following primary pancreatic resection. (2013)
  • Author(s): Hashimoto D, Chikamoto A, Ohmuraya M, Sakata K, Miyake K, Kuroki H, Watanabe M, Beppu T, Hirota M, Baba H.
  • Journal Title: Surg Today Volume: - Issue: 7 Pages: 1313-1320
  • DOI: 10.1007/s00595-013-0708-0
  • Peer Reviewed
• [Journal Article] Pancreaticodigestive anastomosis and the postoperative management strategies to prevent postoperative pancreatic fistula formation after pancreaticoduodenectomy. (2013)
  • Author(s): Hashimoto D, Chikamoto A, Ohmuraya M, Hirota M, Baba H
  • Journal Title: Surg Today Volume: - Issue: 7 Pages: 1207-1213
  • DOI: 10.1007/s00595-013-0662-x
  • Peer Reviewed
• [Journal Article] Serine protease inhibitor Kazal type 1 and epidermal growth factor receptor are expressed in pancreatic tubular adenocarcinoma, intraductal papillary mucinous neoplasm, and pancreatic intraepithelial neoplasia. (2013)
  • Author(s): Ozaki N, Ohmuraya M, Ida S, Hashimoto D, Ikuta Y, Chikamoto A, Hirota M, Baba H.
  • Journal Title: J Hepatobiliary Pancreat Sci.
  • NAID: 10031194618
  • Peer Reviewed
• [Journal Article] c-FLIP maintains tissue homeostasis by preventing apoptosis and programmed necrosis. (2013)
  • Author(s): Piao X
  • Journal Title: Sci Signal Volume: ra93 Issue: 255
  • DOI: 10.1126/scisignal.2003558
  • Peer Reviewed
• [Journal Article] Role of intrapancreatic SPINK1/Spink3 expression in the development of pancreatitis (2012)
  • Author(s): Ohmuraya, M., Sugano, A., Hirota, M., Takaoka, Y. and Yamamura, K
  • Journal Title: Frontiers in Physiol Volume: 3
  • DOI: 10.3389/fphys.2012.00126
  • Peer Reviewed
• [Presentation] オートファジー不全と慢性膵炎発症メカニズムの解明 (2014)
  • Author(s): 大村谷昌樹,坂田和也,高城克暢,松元美由紀,荒木喜美,山村研一
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜、横浜市、神奈川県
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] SPINK insufficiency causes impaired autophagy resulting in chronic pancreatitis. (2014)
  • Author(s): Ohmuraya, M., Sakata, K., Mareninova, O., Notohara, K., Gukovsky, I., Gukovskaya, A.
  • Organizer: 2014 APA/JPS 45th Anniversary Meeting
  • Place of Presentation: Hapuna Beach Prince Hotel、ハワイ島、ハワイ、米国
  • Year and Date: 2014-11-05 – 2014-11-08
• [Presentation] SPINK不全に伴うオートファジーの異常と慢性膵炎の発症 (2014)
  • Author(s): 大村谷昌樹
  • Organizer: 第22回若手膵臓研究会
  • Place of Presentation: 生田神社会館、神戸市、兵庫県
  • Year and Date: 2014-10-22
• [Presentation] オートファジー不全と慢性膵炎発症メカニズムの解明 (2014)
  • Author(s): 大村谷昌樹
  • Organizer: 第23回日本Cell Death学会学術集会
  • Place of Presentation: 東京医科歯科大学、東京都
  • Year and Date: 2014-07-18 – 2014-07-19
• [Presentation] 急性膵炎におけるオートファジー選択的基質p62の役割 (2014)
  • Author(s): 大村谷昌樹,能登原憲司,廣田昌彦
  • Organizer: 第45回日本膵臓学会
  • Place of Presentation: 北九州国際会議場、北九州市、福岡
  • Year and Date: 2014-07-11 – 2014-07-12
• [Presentation] Autophagic cell death by SPINK insufficiency induces chronic inflammation in the pancreas (2013)
  • Author(s): Masaki Ohmuraya, Zhenghua Li, Ken-ichi Yamamura
  • Organizer: 45th Annual Meeting of the European Pancreatic Club
  • Place of Presentation: Kongresshaus, Zurich, Switzerland
• [Presentation] p62/SQSTM1 Is Involved in the Mechanism of Acute Pancreatitis. (2013)
  • Author(s): Masaki Ohmuraya, Kenji Notohara, Ken-ichi Yamamura
  • Organizer: 44th Annual Meeting of the American Pancreatic Association
  • Place of Presentation: Eden Roc Hotel, Miami, FL, USA
• [Presentation] カテプシンDによる腺房細胞内トリプシン活性化機構の解析 (2013)
  • Author(s): 大村谷昌樹、橋本大輔、坂田和也、能登原憲司、馬場秀夫
  • Organizer: 第44回 日本膵臓学会大会
  • Place of Presentation: 仙台国際センター、仙台、宮城
• [Presentation] Chronic inflammation in the pancreas induces pre-cancerous development in mice (2013)
  • Author(s): Masaki Ohmuraya, Kazuya Sakata, Kenji Notohara, Kimi Araki, Hideo Baba, Ken-ichi Yamamura.
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜、神奈川
• [Presentation] オートファジー不全がもたらす慢性膵炎発症メカニズムの解析 (2013)
  • Author(s): Masaki Ohmuraya, Kazuya Sakata, Kenji Notohara, Kimi Araki, Hideo Baba, Ken-ichi Yamamura.
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド、神戸、兵庫
• [Presentation] Insufficient levels of serine protease inhibitor Kazal (SPINK) produced by partial expression of SPINK1 in Spink3 deficient pancreas result in chronic pancreatitis. (2012)
  • Author(s): Masaki Ohmuraya, Kenji Notohara, Kimi Araki, Masahiko Hirota, Ken-ichi Yamamura
  • Organizer: アメリカ消化器病週間2013
  • Place of Presentation: San Diego Convention Center、米国
• [Presentation] Chronic pancreatitis triggers factors promoting pancreatic cancer development (2012)
  • Author(s): Masaki Ohmuraya, Kenji Notohara, Kimi Araki, Hideo Baba, Ken-ichi Yamamura
  • Organizer: 第71回日本癌学会
  • Place of Presentation: ロイトン札幌、札幌市
• [Presentation] ヒトSPINK1遺伝子ノックインマウスの樹立と解析 (2012)
  • Author(s): 大村谷昌樹、能登原憲司、廣田昌彦
  • Organizer: 第43回日本膵臓学会大会
  • Place of Presentation: ホテルメトロポリタン山形、山形市