A transgenic-cloned pig model expressing fluorescent or non-fluorescent modified Plum gene for the evaluation of islet transplantation

• Project/Area Number: 25293279
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: General surgery
• Research Institution: Meiji University
• Principal Investigator: Masaki Nagaya 明治大学, 公私立大学の部局等, 教授 (90329300)
• Co-Investigator(Kenkyū-buntansha): NAGASHIMA HIROSHI 明治大学, 農学部, 教授 (50318664) ; UMEYAMA KAZUHIRO 明治大学, 研究・知財戦略機構, 特任准教授 (70342699) ; WATANABE MASATO 明治大学, 研究・知財戦略機構, 特任講師 (00321688) ; MATSUNARI HITOMI 明治大学, 研究・知財戦略機構, 特任講師 (70639517) ; ARAI YOSHIKAZU 明治大学, 研究・知財戦略機構, 研究推進員 (90614769)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2015: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2013: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
• Keywords: 膵ランゲルハンス島 / トランスジェニックブタ / 膵島移植 / 拒絶 / 移植 / 膵ランゲルハンス糖 / 糖尿病 / クローンブタ / ヒト糖尿病モデル / 赤紫蛍光蛋白Plum / ヒト糖尿病モデルモデル

Outline of Final Research Achievements

A problem with pancreatic islet transplantation for type-1 diabetes patients is the low engraftment rate of islets in the recipient. About 90% of islet transplantations are carried out by intrahepatic transplantation through the portal vein, but a variety of problems occur with this method. Frequent transplants necessary for insulin therapy withdrawal because overcoming the serious problem of transplanted islets will be impossible until new, appropriate transplant method is discovered. If two individual pigs- one that expresses fluorescent proteins, and one that expresses mutated non-fluorescent forms of these proteins- are generated using somatic cell cloning technology, the recipient pig will become immunotolerant to the donor pancreatic islet. This research was undertaken to generate such pigs and to determine appropriate sites for transplantation of islet transplantation.

Research Products

• [Journal Article] A New System to Evaluate the Influence of Immunosuppressive Drugs on Pancreatic Islets Using Epigenetic Analysis in a 3-Dimensional Culture. (2015)
  • Author(s): Nagaya M, Arai Y, Matsunari H, Honda M, Nakano K, Maehara M, Sugimoto N, Kobayashi M, Sakai R, Asano Y, Watanabe M, Umeyama K, Nagashima H.
  • Journal Title: Pancreas. Volume: 44 Issue: 5 Pages: 778-85
  • DOI: 10.1097/mpa.0000000000000366
  • Peer Reviewed
• [Journal Article] Production of transgenic cloned pigs expressing the far-red fluorescent protein monomeric Plum (2015)
  • Author(s): Watanabe M, Kobayashi M, Nagaya M, Matsunari H, Nakano K, Maehara M, Hayashida G, Takayanagi S, Sakai R, Umeyama K, Watanabe N, Onodera M, Nagashima H.
  • Journal Title: Journal of Reproduction and Development Volume: 61 Issue: 3 Pages: 169-177
  • DOI: 10.1262/jrd.2014-153
  • NAID: 130005077737
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed
• [Journal Article] Production of transgenic cloned pigs expressing the far-red fluorescent protein monomeric Plum. (2015)
  • Author(s): 1.Watanabe M, Kobayashi M, Nagaya M, Matsunari H, Nakano K, Maehara M, Hayashida G, Takayanagi S, Sakai R, Umeyama K, Watanabe N, Onodera M, Nagashima H.
  • Journal Title: Journal of Reproduction and Development Volume: -
  • NAID: 130005077737
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Transgenic pigs with pancres-specific expression of green fluorescent protein. (2014)
  • Author(s): 2.Matsunari H, Kobayashi T, Watanabe K, Umeyama K, Nakano K, Kanai T, Matsuda T, Nagaya M, Hara M, Nakauchi H, Nagashima H: Transgenic pigs with pancres-specific expression of green fluorescent protein.
  • Journal Title: Journal of Reproduction and Development Volume: 60 Pages: 230-237
  • Peer Reviewed
• [Journal Article] A conserved rule for pancreatic islet rganization. (2014)
  • Author(s): 3.Hoang D-T, Matsunari H, Nagaya M, Nagashima H, Millis JM, Witkowski P, Periwal V, Hara M, Jo J
  • Journal Title: PLOS ONE Volume: 9 Issue: 10 Pages: e110384-e110384
  • DOI: 10.1371/journal.pone.0110384
  • Peer Reviewed / Open Access
• [Journal Article] Transgenic pigs with pancreas specific expression of green fluorescent protein. (2014)
  • Author(s): Matsunari H, Kobayashi T., Watanabe M, Umeyama K, Nakano K, Kanai T, Matsuda T, Nagaya M, Hara M, Nakauchi H, Nagashima H
  • Journal Title: J Reprod Dev Volume: [Epub ahead of print]
  • NAID: 130003390840
  • Peer Reviewed
• [Journal Article] Generation of interleukin-2 receptor gamma gene knockout pigs from so matic cells genetically modified by zinc finger nuclease-encoding mRNA. (2013)
  • Author(s): Watanabe M, Nakano K, Matsunari H, Matsuda T, Maehara M, Kanai T, Kobayashi M, Matsumura Y, Sakai R, Kuramoto M, Hayashida G, Asano Y, Takayanagi S, Arai Y, Umeyama K., Nagaya M, Hanazono Y, Nagashima H
  • Journal Title: PLOS ONE Volume: 8 Issue: 10 Pages: e76478-e76478
  • DOI: 10.1371/journal.pone.0076478
  • Peer Reviewed
• [Journal Article] Production of diabetic offspring using cryopreserved epididymal sperm by in vitro fertilization and intrafallopian insemination techniques in transgenic pigs. (2013)
  • Author(s): Umeyama K, Honda K, Matsunari H, Nakano K, Hidaka T, Sekiguchi K, Mochizuki H, Takeuchi Y, Fujiwara T, Watanabe M, Nagaya M, Nagashima H.
  • Journal Title: J Reprod Dev. Volume: 59 Pages: 599-603
  • NAID: 130003361059
  • Peer Reviewed
• [Presentation] ブタにおけるキメラ胚の発達動態 (2015)
  • Author(s): 1.畑江将太, 中野和明, 松成ひとみ, 内倉鮎子, 浅野吉則, 武石透輝, 渡邊將人, 梅山一大, 長屋昌樹, 長嶋比呂志
  • Organizer: 第108回日本繁殖生物学会
  • Place of Presentation: 宮崎
  • Year and Date: 2015-09-17
• [Presentation] Blastocyst complementation restore apancreatic phenotype of pdx1 pigs. (2015)
  • Author(s): 1.Matsunari H, Watanabe M, Nakano K, Uchikura A, Asano Y, Umeyama K, Yamaguchi T, Nagaya M, Nakauchi H, Nagashima H
  • Organizer: International Society for Stem Cell Research 2015 Annual Meeting
  • Place of Presentation: Stockholm, Sweden.
  • Year and Date: 2015-06-24
  • Int'l Joint Research
• [Presentation] Production efficiency of gene knockout pigs using genome editing and somatic cell cloning (2015)
  • Author(s): 1.Matsunari H, Watanabe M, Nakano K, Uchikura A, Asano Y, Hatae S, Takeishi T, Umeyama K, Nagaya M, Miyagawa S, Hanazono Y, Nakauchi H, Nagashima H
  • Organizer: 41th Annual Conference of the International Embryo Transfer Society
  • Place of Presentation: Versailles, France
  • Year and Date: 2015-01-10 – 2015-01-13
• [Presentation] Transmission of lethal phenotype in a Mendelian fashion by genetically modified pigs that underwent blastocyst complementatio (2014)
  • Author(s): 2.Matsunari H, Watanabe M, Umeyama K, Nakano K, Nagaya M, Nakauchi H, Nagashima H
  • Organizer: World Congress of Reproductive Biology
  • Place of Presentation: Edinburgh, UK
  • Year and Date: 2014-09-02 – 2014-09-04
• [Presentation] 糖尿病発症トランスジェニックブタ凍結精子の体外受精能の評価. (2014)
  • Author(s): 1.倉本桃子, 小林美里奈, 林田豪太, 松村幸奈, 松成ひとみ, 中野和明, 鞍本友香, 一山琴世, 浅野吉則, 内倉鮎子, 渡邊將人, 梅山一大, 長屋昌樹, 長嶋比呂志
  • Organizer: 第107回日本繁殖生物学会大会
  • Place of Presentation: 帯広
  • Year and Date: 2014-08-21 – 2014-08-24
• [Presentation] 遠赤色蛍光タンパクmonomeric Plum を全身発現するトランスジェニック・クローンブタの作出 (2014)
  • Author(s): 2.小林美里奈, 渡邊將人, 松成ひとみ, 中野和明, 林田豪太, 坂井理恵子, 松村幸奈, 倉本桃子, 梅山一大, 渡辺信之, 小野寺雅史, 長屋昌樹, 長嶋比呂志
  • Organizer: 第107回日本繁殖生物学会大会
  • Place of Presentation: 帯広
  • Year and Date: 2014-08-21 – 2014-08-24
• [Presentation] 細胞周期可視化蛍光プローブFucci導入トランスジェニック・クローンブタの作出 (2014)
  • Author(s): 3.渡邊將人, 林田豪太, 松成ひとみ, 中野和明, 小林美里奈, 松村幸奈, 倉本桃子, 坂井理恵子, 浅野吉則, 内倉鮎子, 新井良和, 梅山一大, 長屋昌樹, 阪上‐沢野朝子, 宮脇敦史, 長嶋比呂志
  • Organizer: 第107回日本繁殖生物学会大会
  • Place of Presentation: 帯広
  • Year and Date: 2014-08-21 – 2014-08-24
• [Presentation] Development of apancreatic pig by genome editing and its application in organ regeneration research (2014)
  • Author(s): 3.Matsunari H, Watanabe M, Nakano K, Uchikura A, Asano Y, Kobayashi M, Umeyama K, Nagaya M, Nakauchi H, Nagashima H
  • Organizer: Swine in Biomedical Research International Conference
  • Place of Presentation: Raleigh, USA
  • Year and Date: 2014-07-06 – 2014-07-08
• [Presentation] In vivo exogenic organ generation with organogenesis-disabled cloned pigs as a platform. (2014)
  • Author(s): Matsunari H, Nakano K, Kanai T, Matsuda T, Maehara M, Watanabe M, Umeyama K, Nagaya M, Nakauchi H, Nagashima H
  • Organizer: 40th Annual Conference of the International Embryo Transfer Society
  • Place of Presentation: Reno, USA.
• [Presentation] Generation and characterization of transgenic-cloned pigs expressing the far-red fluorescent protein monomeric plum. (2014)
  • Author(s): Kobayashi M, Watanabe M, Matsunari H, Nakano K, Kanai T, Hayashida G, Matsumura Y, Kuramoto M, Sakai R, Arai Y, Umeyama K, Watanabe N, Onodera M, Nagaya M, Nagashima H.
  • Organizer: 40th Annual Conference of the International Embryo Transfer Society
  • Place of Presentation: Reno, USA.
• [Presentation] 膵臓欠損ブタ胎仔を用いた外来性細胞由来膵臓形成の試み. (2014)
  • Author(s): 松成ひとみ、浅野吉則、小林美里奈、内倉鮎子、渡邊將人、梅山一大、長屋昌樹、中内啓光、長嶋比呂志.
  • Organizer: 第13回日本再生医療学会総会.
  • Place of Presentation: 京都.
• [Presentation] Application of blastocyst complementation to development of genetically modified pigs for xenotransplantation. (2013)
  • Author(s): Nakano K, Matsunari H, Matsuda T, Kanai T, Hayashida G, Matsumura K, Kobayashi M, Kuramoto M, Asano Y, Sakai R, Uchikura A, Arai Y, Watanabe M, Umeyama K, Nagaya M, Nagashima H.
  • Organizer: 12th Congress of International Xenotransplantation Association (IXA).
  • Place of Presentation: Osaka, Japan.
• [Presentation] In vivo generation of exogenic pancreas in apancreatic cloned pigs via blastocyst complementation. (2013)
  • Author(s): Matsunari H, Nakano K, Kanai T, Matsuda T, Maehara M, Watanabe M, Umeyama K, Nagaya M, Nakauchi H, Nagashima H.
  • Organizer: 12th Congress of International Xenotransplantation Association (IXA).
  • Place of Presentation: Osaka, Japan.
• [Presentation] ブタ卵におけるmRNA injection法を用いたZinc Finger Nucleasesによる遺伝子ノックアウト. (2013)
  • Author(s): 松田泰輔, 渡邊將人, 中野和明, 松成ひとみ, 小林美里奈, 林田豪太, 倉本桃子, 金井貴博, 山口智之, 中内啓光, 長屋昌樹, 長嶋比呂志.
  • Organizer: 第36回日本分子生物学会.
  • Place of Presentation: 神戸.
• [Presentation] Zinc finger nuclease発現mRNAによるIL2RG遺伝子ノックアウトブタの作出. (2013)
  • Author(s): 渡邊將人, 中野和明, 松成ひとみ, 松田泰輔, 金井貴博, 小林美里奈, 松村幸奈, 坂井理恵子, 倉本桃子, 林田豪太, 浅野吉則, 高柳就子, 新井良和, 梅山一大, 長屋昌樹, 豊 花, 長嶋比呂志
  • Organizer: 第36回日本分子生物学会.
  • Place of Presentation: 神戸.
• [Presentation] 異種移植研究における遺伝子改変ブタの作出への人工生殖技術の利用. (2013)
  • Author(s): 坂井理恵子, 中野和明, 松成ひとみ, 新井良和, 渡邊將人, 梅山一大, 佐原寿史, 山田和彦, 長屋昌樹, 宮川周士, 長嶋比呂志.
  • Organizer: 第1回日本先進医工学ブタ研究会.
  • Place of Presentation: 大阪.
• [Presentation] 遺伝子改変クローンブタ開発における人工生殖技術を利用した遺伝的バックグラウンドの更新. (2013)
  • Author(s): 坂井理恵子, 中野和明, 松成ひとみ, 新井良和, 渡邊將人, 梅山一大, 長屋昌樹, 宮川周士, 長嶋比呂志.
  • Organizer: 第16回日本異種移植研究会.
  • Place of Presentation: 大阪.
• [Presentation] 細胞周期可視化蛍光プローブFucci を発現するブタ体細胞核移植胚の作出. (2013)
  • Author(s): 林田豪太, 渡邊将人, 松成ひとみ, 中野和明, 金井貴博, 小林美里奈, 松村幸奈, 倉本桃子, 坂井理恵子, 浅野吉則, 内倉鮎子, 前原美樹, 新井良和, 梅山一大, 長屋昌樹, 沢野朝子, 宮脇敦史, 長嶋比呂志.
  • Organizer: 第106回日本繁殖生物学会大会.
  • Place of Presentation: 東京.
• [Presentation] ガラス化保存された体外成熟／体外受精胚を用いた糖尿病モデル遺伝子改変ブタの作出. (2013)
  • Author(s): 松村幸奈, 前原美樹, 本田香澄, 林田豪太, 倉本桃子, 中野和明, 松成ひとみ, 小林美里奈, 内倉鮎子, 浅野吉則, 渡邊将人, 梅山一大, 長屋昌樹, 長嶋比呂志.
  • Organizer: 第106回日本繁殖生物学会大会.
  • Place of Presentation: 東京.
• [Remarks] 明治大学バイオリソース研究国際インスティチュート
  • URL: http://muiibr.com/institute/kenkyuchizai.html
• [Remarks] 明治大学バイオリソース研究国際インスティチュート
  • URL: http://muiibr.com/