Establishment of Transgenic Pig for Organ Xenotransplantation

用于器官异种移植的转基因猪的建立

• 批准号: 06557068
• 负责人: TAKAGI Hiroshi
• 金额: $ 10.37万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557068/
• 关键词: Xenotransplantaion; Transgenic pig; Complement regulatory factor; Xenoantigen; Gene engineering; 補体制御因子遺伝子; トランスジェニック動物

In organ xenotransplantetion, hyperacute rejection, induced by natural antibody, xenoantigen, and complement, occurs in a few minutes or hours after reperfusion. In this study, based on the in vitro study, in which the double transduction of complement regulatory factors (DAF and HRF20) is more effective than the single transduction on the xenogeneic cells to inhibit complement-dependent cytotoxicity, double transgenic mice with DAF and HRF20 genes were established. Then the construction of transgenic pig with DAF,HRF20, and membrane cofactor protein (MCP) was studied. To control the expression of major xenoantigen GalalphaGal, double knockout of alpha (1,3) GT gene on embryonic stem (ES) cells was examined. As another strategy to control GalalphaGal, it was demonstrated that the transduction of alpha (1,2) FT gene to the xenogenic cell inhibited the GaalphaaGal expression and complement-dependent cytotoxicity, and the transgenic pig with alpha (1,2) FT gene was established. Thus it is concluded that the construction of transgenic pigs with complement regulatory factor genes and alpha (1,2) FT gene is useful for the successful clinical xenotransplantation.

在异种器官移植中，由天然抗体、异种抗原和补体诱导的超急性排斥反应在再灌注后几分钟或几小时内发生。本研究在体外研究补体调节因子（HRF 20和HRF 20）双转导对异种细胞的补体依赖性细胞毒性抑制作用优于单转导的基础上，建立了HRF 20和HRF 20双转基因小鼠。在此基础上，研究了转HRF、HRF 20和膜辅因子蛋白（MCP）基因猪的构建。为了控制主要异种抗原GalalphaGal的表达，检查了胚胎干（ES）细胞上α（1，3）GT基因的双敲除。作为另一种控制GalalphaGal的策略，证明了将α（1，2）FT基因转导到异种细胞中抑制了GalalphaGal的表达和补体依赖的细胞毒性，并建立了α（1，2）FT基因转基因猪。因此，构建含有补体调节因子基因和α（1，2）FT基因的转基因猪，为临床异种移植的成功提供了实验依据。

项目成果

S.Hayashi,et al.: "Effect of antisence ribozyme to pit α(1,3) galactosyl transferase gene on the expression of Gal α(1,3)Gal epitope" Transplantation Proceedings. (in press).

S. Hayashi 等人：“凹坑 α(1,3) 半乳糖基转移酶基因的反义核酶对 Gal α(1,3)Gal 表位表达的影响”移植论文集（出版中）。

Shuji Hayashi: "Synergistic effect of donor pretreatment using FK506 in hamster to rat cardiac xenotransplantation" Transplantation Proceedings. 26. 1284 (1994)

Shuji Hayashi：“在仓鼠至大鼠心脏异种移植中使用 FK506 供体预处理的协同效应”移植论文集。

Y.Namii, et al: "Tetrahydropyranyladriamycin with FK506 combination therapy for hamster-to-rat concordant xenotransplantation" Transpl Proc. (in press).

Y.Namii 等人：“四氢吡喃拉霉素与 FK506 联合疗法用于仓鼠-大鼠一致异种移植”Transpl Proc。

S.Hayashi, et al: "Protection of guinea pig cells transfected with 512 gene from complement dependent cytolysis of rat" Transpl Proc. (in press).

S.Hayashi 等人：“用 512 基因转染的豚鼠细胞免受大鼠补体依赖性细胞溶解的保护”Transpl Proc。

S.Hayashi, et al: "Evidence that double transfection to xenoendothelial cells using GPI-anchoring complement regulatory factor (DAF and HRF20) genes is useful for the inhibition of human complement-mediated cytolysis" Xenotransplantation. 2. 259-294 (1995

S.Hayashi 等人：“有证据表明使用 GPI 锚定补体调节因子（DAF 和 HRF20）基因双重转染异种内皮细胞可用于抑制人补体介导的细胞溶解作用”异种移植。