Gene therapy for organ grafts using rapid injection of naked DNA: Application to the rat liver
Gene therapy for organ grafts using rapid injection of naked DNA: Application to the rat liver
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DOI：
10.1097/01.tp.0000118404.66106.e8
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发表时间：
2004-04-15
期刊：
影响因子：
6.2
通讯作者：
Kobayashi, E
中科院分区：
文献类型：
--
作者：
Inoue, S;Hakamata, Y;Kobayashi, E
背景我们建立了一种快速注射裸DNA靶向肝脏的非病毒基因转移方法，并将其应用于大鼠肝移植模型。方法.使用近交的Dark Agglutinus和刘易斯大鼠。为了测试全身或局部（基于导管）注射的功效和副作用，注射不同体积的含有编码β-半乳糖苷酶（IacZ）的裸DNA的磷酸盐缓冲盐水。荧光素酶表达后进行非侵入性成像，细胞毒性T淋巴细胞抗原4-免疫球蛋白（CTLA 4 Ig）蛋白进行了功能测试同种异体心脏移植。然后在大鼠辅助性肝移植（ALT）和原位肝移植（奥尔特）中测试基因转移。在辅助性肝移植模型中评估基因转移的时机，并使用在2天前转移荧光素酶或CTLA 4 Ig基因的肝移植物进行奥尔特。LacZ以体积依赖性方式广泛表达;然而，大体积常常诱导受体死亡。在将CTLA 4 Ig cDNA局部递送至肝脏后，随着CTLA 4 Ig血清水平的增加，Dark Agglutination心脏移植物的存活时间延长。在捐赠时注射裸DNA的肝脏移植物不能存活，但在基因转移后2天移植的肝脏存活。结果表明，OLT大鼠体内成功表达了荧光素酶和CTLA 4 Ig。我们成功地将非病毒流体动力学基因转移方法应用于大鼠肝脏，并显示了其在肝脏移植中的潜力。在器官捐献当天进行该手术时移植失败的高发生率是临床应用中需要克服的潜在限制。
Background. We developed a nonviral gene transfer method using rapid injection of naked DNA targeting the liver and applied it in a rat model of liver transplantation. Methods. Inbred Dark Agouti and Lewis rats were used. To test the efficacy and adverse effects of systemic or local (catheter-based) injection, different volumes of phosphate-buffered saline containing naked DNA encoding beta-galactosidase (IacZ) were injected. Luciferase expression was followed by non-invasive imaging, and a cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4Ig) protein was tested functionally by allogenic heart transplantation. Gene transfer was then tested in rat auxiliary liver transplantation (ALT) and orthotopic liver transplantation (OLT). The timing of gene transfer was evaluated in the auxiliary liver transplantation model, and OLT was performed using a liver graft to which luciferase or the CTLA4Ig gene was transferred 2 days before.Results. LacZ was expressed extensively in a volume-dependent manner; however, a large volume often induced recipient death. After local delivery of CTLA4Ig cDNA to the liver, survival of Dark Agouti heart grafts lengthened with increased CTLA4Ig serum levels. Liver grafts injected with naked DNA at the time of donation did not survive, but livers grafted 2 days after gene transfer survived. Successful expression of luciferase and production of CTLA4Ig were finally confirmed in the rat that underwent OLT.Conclusions. We successfully applied a nonviral hydrodynamic gene transfer method to the rat liver and showed its potential in liver grafting. The high incidence of graft failure when this procedure is performed on the day of organ donation is a potential limitation that needs to be overcome in clinical application.