GalT-knockout pig to monkey xenogeneic kidney and islet transplantation

GalT 基因敲除猪猴异种肾和胰岛移植

• 批准号: 19209043
• 负责人: YAMADA Kazuhiko
• 金额: $ 31.62万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-19209043/
• 关键词: 異種移植; ブタ・サル間移植; 腎移植; 膵島移植; GalKOブタ; 遺伝子導入; hCD47; 免疫寛容

Due largely to improvements in immunosuppressive therapies, organ transplantation has become the primary cure for end-stage organ failure in the United States, Europe, as well as Japan. However, the current shortage of donor organs is a critical problem. One attractive solution is xenotransplantation utilizing miniature swine donor organs. However, because human blood contains native antibodies (NAb) directed toward a constitutively expressed pig cell surface carbohydrate called galactose-alpha-1,3-galactose (Gal), pig-to-human xenotransplantation initially resulted in hyper-acute (graft loss in <1 day) humoral rejection. With this grant support in the past four years, we have made two major achievements, neither of which had previously been accomplished in Japan. Achievement 1 (first domestic success) : Using nuclear transfer from somatic nuclei of GalT-KO miniature swine, obtained from the TBRC at Harvard Medical School, we and our colleagues, as a collaborative effort, succeeded in … More producing GalT-KO miniature swine. We then transplanted GalT-KO kidneys from those pigs into cynomologous monkeys to confirm that hyperacute rejection was avoidable. Rejection did not occur for 15 days after xenotransplantation. Achievement 2 (first domestic success) : We successfully isolated islets from miniature swine and, following xenogeneic transplantation of these islets, we observed maintenance of normal blood glucose levels up to two months in monkeys. In addition, we have explored the mechanisms of rejection in xenotransplantation. In particular, our results utilizing GalT-KO swine suggested that CD59 is involved in development of accommodation. Moreover, our studies have suggested immune-regulatory/protective effects by (1) transgene human CD47 onto porcine cells and (2) administration of soluble thrombomodulin or hepatocyte growth factor. Using the support of this grant, we have developed new immunosuppressive regimens for renal/islet xenotransplantation, including these reagents. Less

在美国、欧洲和日本，主要由于免疫抑制疗法的改进，器官移植已经成为终末期器官衰竭的主要治疗方法。然而，目前捐赠器官的短缺是一个严重的问题。一个有吸引力的解决方案是利用小型猪供体器官进行异种移植。然而，由于人类血液中含有针对猪细胞表面碳水化合物的天然抗体(NAB)，这种碳水化合物被称为半乳糖-α-1，3-半乳糖(Gal)，猪到人的异种移植最初会导致超急性(移植物丢失1天内)体液排斥反应。在过去四年的赠款支持下，我们取得了两项重大成就，这两项成就以前都不是在日本实现的。成果1(国内首例成功)：利用哈佛医学院生物医学研究中心的GALT-KO小型猪体细胞核移植，我们和同事合作，成功地进行了…。生产更多的Galt-KO小型猪。然后，我们将这些猪的Galt-KO肾脏移植到染色体正常的猴子体内，以证实超急性排斥反应是可以避免的。异种移植后15天内未发生排斥反应。成果2(国内首例成功)：我们成功地从小型猪中分离出胰岛，在异种移植这些胰岛后，我们观察到猴子的正常血糖水平维持了长达两个月。此外，我们还对异种移植排斥反应的发生机制进行了探讨。特别是，我们利用Galt-KO猪的结果表明CD59参与了调节的发展。此外，我们的研究表明，(1)将人CD47转基因到猪细胞上，以及(2)给予可溶性血栓调节蛋白或肝细胞生长因子，具有免疫调节/保护作用。利用这笔赠款的支持，我们开发了用于异种肾/胰岛移植的新的免疫抑制方案，包括这些试剂。较少

项目成果

Necessity and possibility of serial cloning in development of advanced genetically engineered pigs for xenotransplantation.

连续克隆在开发用于异种移植的先进基因工程猪的必要性和可能性。

• 发表时间: 2010
• 作者: Nagashima H;et al.
• 通讯作者: et al.

Hepatocyte Growth Factor regimen in a life supporting xenogeneic islet transplantation CLAWN miniature swine-to -cynomologous monkey model.

生命支持异种胰岛移植 CLAWN 小型猪至食蟹猴模型中的肝细胞生长因子方案。

• 发表时间: 2010
• 作者: Yamada K;Nishimura H;Sahara H.;et al
• 通讯作者: et al

レクチンブロット法によるαGa1-knockout ブタの糖鎖抗原の解析

通过凝集素印迹分析 αGa1 敲除猪碳水化合物抗原

• 发表时间: 2009
• 作者: 山本亜紀;徐恒傑;武石俊作;山田正雄;三善英知;池田孔佑;松成ひとみ;長嶋比呂志
• 通讯作者: 長嶋比呂志

α1, 3ガラクトース転移酵素遺伝子ダブルノックアウトブタの体細胞クローニングにおける Scriptaid の効果

Scriptaid对α1,3半乳糖基转移酶基因双敲除猪体细胞克隆的影响

• 发表时间: 2010
• 作者: 松成ひとみ;池澤有加;渡邊将人;梅山一大;中野和明;藤原主;竹内靖浩;本田香澄;前原美樹;高柳就子;山田和彦;宮川周士;長嶋比呂志
• 通讯作者: 長嶋比呂志

Knockout of exogenous EGFP gene in porcine somatic cells using zink-finger nucleases.

使用锌指核酸酶敲除猪体细胞中的外源 EGFP 基因。

• 发表时间: 2010
• 期刊: Biochemical and Biophysical Research Communications
• 影响因子: 3.1
• 作者: Watanabe M;Umeyama K;Matsunari H;Takayanagi S;Haruyama E;Nakano K;Fujiwara T;Ikezawa Y;Nakauchi H;Nagashima H
• 通讯作者: Nagashima H