Basic research for hematopoietic stem cell transplantation before birth

• Project/Area Number: 24591429
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Jichi Medical University
• Principal Investigator: HANAZONO Yutaka 自治医科大学, 医学部, 教授 (70251246)
• Research Collaborator: NAGAO Yosikazu 宇都宮大学, 農学部, 教授 (70291953) ; ABE Tomoyuki 自治医科大学, 医学部 分子病態治療研究センター, 助教 (20610364)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 造血幹細胞移植 / 出生前治療 / 先天性免疫不全症 / 大型動物 / ヒツジ / 子宮内移植 / 幹細胞移植 / 胎児 / 前処置

Outline of Final Research Achievements

X-linked severe combined immunodeficiency has been successfully treated in US and European countries by in utero transplantation (IUT) of hematopoietic stem cells; however, the engraftment efficiency has not necessarily been high enough for other diseases in which graft cells do not have growth advantage. From a series of experiments in the setting of sheep IUT, we have shown that the engraftment efficiency can be improved by the following methods: (1) Pre-conditioning of fetuses with busulfan, (2) transient forced expression of the HoxB4 gene in donor hematopoietic stem cells, and (3) post-conditioning with cytokines.

Research Products

• [Journal Article] A long-term follow-up study on the engraftment of human hematopoietic stem cells in sheep (2014)
  • Author(s): Abe, T., Hanazono, Y., Nagao, Y.
  • Journal Title: Exp. Anim. Volume: 63 Pages: 475-481
  • NAID: 130004701642
  • Peer Reviewed
• [Journal Article] Generating porcine chimeras using inner cell mass cells and parthenogenetic preimplantation embryos (2013)
  • Author(s): Nakano, K., Watanabe, M., Matsunari, H., Matsuda, T., Honda, K., Maehara, M., Kanai, T., Hayashida, G., Kobayashi, M., Kuramoto, M., Arai, Y., Umeyama, K., Fujishiro, S.-H., Mizukami, Y., Nagaya, M., Hanazono, Y., Nagashima, H.
  • Journal Title: PLOS ONE Volume: 8
  • Peer Reviewed
• [Journal Article] 大型動物を用いた幹細胞研究 (2013)
  • Author(s): 花園豊
  • Journal Title: 臨床血液 Volume: 54 Pages: 329-335
  • NAID: 10031171335
  • Peer Reviewed
• [Journal Article] DNA methylation profiles provide a viable index for porcine pluripotent stem cells (2013)
  • Author(s): Arai, Y., Ohgane, J., Fujishiro, S.-H., Nakano, K., Matsunari, H., Watanabe, M., Umeyama, K., Azuma, D., Uchida, N., Sakamoto, N., Makino, T., Yagi, S., Shiota, K., Hanazono, Y., Nagashima, H.
  • Journal Title: Genesis Volume: 51 Pages: 763-776
  • Peer Reviewed
• [Journal Article] Generation of interleukin-2 receptor gamma gene knockout pigs from somatic 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
  • Peer Reviewed
• [Journal Article] Maternal administration of busulfan before in utero transplantation of human hematopoietic stem cells enhances engraftments in sheep (2012)
  • Author(s): Abe T, Masuda S, Tanaka Y, Nitta S, Kitano Y, Hayashi S, Hanazono Y, Nagao Y
  • Journal Title: Exp Hematol Volume: 40(6) Issue: 6 Pages: 436-444
  • DOI: 10.1016/j.exphem.2012.01.018
  • Peer Reviewed
• [Presentation] ヒツジ子宮内移植系におけるヒト造血細胞の生着促進・増幅技術の開発 (2015)
  • Author(s): 阿部朋行
  • Organizer: 第17回日本異種移植研究会
  • Place of Presentation: 栃木
  • Year and Date: 2015-03-14
• [Presentation] ヒツジ子宮内移植系におけるヒト造血細胞の生着・増幅技術の開発 (2014)
  • Author(s): 阿部朋行
  • Organizer: 第61回日本実験動物学会総会
  • Place of Presentation: 札幌
  • Year and Date: 2014-05-15 – 2014-05-17
• [Presentation] 幹細胞治療研究における医学と獣医学の連携 (2014)
  • Author(s): 花園豊
  • Organizer: 平成25年度獣医学術学会年次大会
  • Place of Presentation: 幕張
  • Invited
• [Presentation] ヒツジ子宮内移植系によるヒト造血幹細胞の定量評価の試み (2014)
  • Author(s): 阿部朋行、長尾慶和、柳瀬公秀、ボラジギン・サラントラガ、緒方和子、山口美緒、花園豊
  • Organizer: 第36回 日本造血細胞移植学会
  • Place of Presentation: 沖縄
• [Presentation] カニクイザル初期胚を用いたES細胞の特性に関する検討 (2013)
  • Author(s): 下澤律浩、藤城修平、水上喜久、阿部朋行、花園豊
  • Organizer: 第54回日本卵子学会
  • Place of Presentation: 東京
• [Presentation] 実物としてのマウス多能性幹細胞DNAメチル化プロフィールに基づく幹細胞評価: ブタiPS細胞を例として (2013)
  • Author(s): 新井良和、大鐘潤、藤城修平、中野和明、塩田邦郎、花園豊、長嶋比呂志
  • Organizer: 第7回日本エピジェネティクス研究会年会
  • Place of Presentation: 奈良
• [Presentation] 再生医学研究：臨床応用をめざして (2013)
  • Author(s): 花園豊
  • Organizer: 第77回日本皮膚科学会東部支部学術大会
  • Place of Presentation: 大宮
• [Presentation] Porcine iPS (2013)
  • Author(s): Hanazono, Y.
  • Organizer: 12th Congress of International Xenotransplantation Association
  • Place of Presentation: 大阪
  • Invited
• [Presentation] Factors influencing engraftment of monkey embryonic stem cells in sheep after xenogeneic in utero transplantation (2013)
  • Author(s): Nagao, Y., Abe, T., Tanaka, Y., Sasaki, K., Masuda, S., Sarentonglaga, B., Ogata, K., Yamaguchi, M., Hayashi, S., Kitano, Y., Hanazono, Y.
  • Organizer: 12th Congress of International Xenotransplantation Association
  • Place of Presentation: 大阪
• [Presentation] Long-term comparative study on the engraftment of human hematopoietic stem cells in sheep after xenogeneic in utero transplantation (2013)
  • Author(s): Abe, T., Masuda, S., Sarentonglaga, B., Ogata, K., Yamaguchi, M., Hayashi, S., Nagao, Y., Hanazono, Y.
  • Organizer: 12th Congress of International Xenotransplantation Association
  • Place of Presentation: 大阪
• [Presentation] Evaluation of porcine induced pluripotent stem cells based on the DNA methylation profile of mouse embryonic stem cell-specific hypomethylated loci (2013)
  • Author(s): Arai, Y., Ohgane, J., Fujishiro, S., Nakano, K., Matsunari, H., Watanabe, M., Umeyama, K., Azuma, D., Uchida, N., Sakamoto, N., Makino, T., Yagi, S., Shiota, K., Hanazono, Y., Nagashima, H.
  • 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、花園豊、長嶋比呂志
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] ヒツジ胎子内におけるサルES細胞の生着、造血系分化およびテラトーマ形成に及ぼす要因 (2012)
  • Author(s): 新田卓, 阿部朋行, 田中裕次郎, 佐々木京子, 増田茂夫, ボラジギン・サラントラガ, 林聡, 北野良博,花園豊, 長尾慶和
  • Organizer: 第105回日本繁殖生物学会
  • Place of Presentation: 茨城
• [Presentation] Possible factors for engraftment of monkey embryonic stem cells after in utero transplantation into sheep fetuses (2012)
  • Author(s): Nagao, Y., Abe, T., Tanaka, Y., Sasaki, K., Masuda, S., Nitta, S., Sarentonglaga, B., Hayashi, S., Kitano, Y., Hanazono, Y.
  • Organizer: 17th International Congress on Animal Reproduction
  • Place of Presentation: Vancouver, Canada
• [Presentation] Long-term follow-up on the engraftment of human HSCs in sheep after in utero transplantation (2012)
  • Author(s): Hanazono, Y., Abe, T., Nitta, S., Masuda, S., Hayashi, S., Nagao, Y.
  • Organizer: International Society for Hematology and Stem Cells 41st Annual Scientific Meeting
  • Place of Presentation: Amsterdam, Netherland
• [Presentation] HoxB4-transduction of human HSC results in long-term engraftment compared to busulfan-conditioning in sheep after in utero transplantation (2012)
  • Author(s): Abe, T., Nitta, S., Masuda, S., Hayashi, S., Nagao, Y., Hanazono, Y.
  • Organizer: The 18th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: 熊本
• [Presentation] Long-term comparative study on the engraftment of human hematopoietic stem cells in sheep (2012)
  • Author(s): Abe, T., Nitta, S., Masuda, S., Hayashi, S., Nagao, Y., Hanazono, Y
  • Organizer: International Society for Stem Cell Research 10th Annual Meeting
  • Place of Presentation: 横浜
• [Presentation] HoxB4-transduction of human HSC results in longer-term engraftment compared to busulfan-conditioning in sheep after in utero transplantation (2012)
  • Author(s): Abe, T., Nitta, S., Masuda, S., Hayashi, S., Nagao, Y., Hanazono, Y.
  • Organizer: 第10回 幹細胞シンポジウム
  • Place of Presentation: 淡路島
• [Presentation] ヒツジにおけるヒト造血細胞長期生着の条件検討 (2012)
  • Author(s): 阿部朋行，新田卓，増田茂夫，林聡，長尾慶和，花園豊
  • Organizer: 日本実験動物科学・技術九州2012
  • Place of Presentation: 別府
• [Presentation] ヒト化ヒツジの作製を目指して
  • Author(s): 阿部朋行、花園豊
  • Organizer: 第60回日本実験動物学会総会ワークショップ
  • Place of Presentation: つくば市
• [Presentation] Human-to-Animal Reversed Xenogeneic Transplantation For Producing Human Blood in Animals
  • Author(s): Hanazono, Y.
  • Organizer: Joint Meeting of the 2nd Symposium of the East Asia Xenotransplantation Association (EAXA) /the 16th Japan Xenotransplantation Association (JXA)
  • Place of Presentation: 大阪国際会議場（大阪）
  • Invited
• [Presentation] ヒツジ子宮内異種移植(I)：生着条件の検討
  • Author(s): 長尾慶和、阿部朋行、柳瀬公秀、ボラジギン・サラントラガ、緒方和子、山口美緒、林聡、花園豊
  • Organizer: 第16回日本異種移植研究会
  • Place of Presentation: 大阪国際会議場（大阪）
• [Presentation] ヒツジ子宮内異種移植(II)：長期間の造血再構築
  • Author(s): 阿部朋行、長尾慶和、柳瀬公秀、ボラジギン・サラントラガ、緒方和子、山口美緒、林聡、花園豊
  • Organizer: 第16回日本異種移植研究会
  • Place of Presentation: 大阪国際会議場（大阪）
• [Presentation] マウスからヒトへ：ブタを利用する橋渡し研究
  • Author(s): 花園豊
  • Organizer: 第1回日本先進医工学ブタ研究会
  • Place of Presentation: 大阪
  • Invited