Xenotransplantation of ex vivo gene trandected cardiomyocytes

离体基因转染心肌细胞的异种移植

基本信息

批准号：
11470277
负责人：
TANIGUCHI Shigeki
金额：
$ 8.13万
依托单位：
Nara Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

We examined xenotransplantion of fetal murine cardiomyocytes to rats, which was made myocardial infarction with coronary artery ligated. In the infracted area, transplanted fetal cardiomyocytes was necrosis due to ischemia, but are alive without hyper-acute rejection in the border zone of normal and infracted area. In addition a little immuno-suppressive agent (cyclosporine 5mg/kg) made donor cells alive over thirty days. We think xenotransplantation of fetal cardiomyocytes are useful for therapy of heart failure.The gene tranduction technologies have greatly expand the potential feasibility of immunotheapy against various disorders including malignancy. A critical impediment of non-viral vector system is the relatively poor Transduction/expression efficiency, which interrupts their real applications to clinical usage. However, the Epstein-Barr virus(EBV)-based episomal plasmid vectors in combination with various delivery vehicles bring very-strong expression in variety of human cells. We tried gene transduction of interferon-β gene to tumor cells with this vector system, and examined anti-tumor effect of gene manipulation. Remarkable suppression of tumor cells growth was obtained by transfection with the EBV-based vector, while transfer of same gene by a conventional Plasmid vector resulted in moderate suppressive effect. Non-viral means equipped with EBV-based plasmid vector offer novel strategies of genetic manipulation applicable to immuno-gene therapy.

我们对小鼠胎儿心肌进行异种移植，结扎冠状动脉制成心肌梗死，移植的胎儿心肌细胞因缺血坏死，但正常与梗死交界处仍存活，未出现超急性排斥反应。另外一点免疫抑制剂（环孢菌素5mg/kg）使供体细胞存活超过30天。胎儿心肌细胞可用于心力衰竭的治疗。基因转导技术极大地扩展了针对包括恶性肿瘤在内的多种疾病的免疫治疗的潜在可行性。非病毒载体系统的一个关键障碍是相对较差的转导/表达效率，这阻碍了其真正的效果。然而，基于 Epstein-Barr 病毒（EBV）的附加型质粒载体与各种载体结合，在多种人类细胞中带来了非常强的表达。用该载体系统将干扰素-β基因转入肿瘤细胞，并检查了基因操作的抗肿瘤效果，通过基于EBV的载体转染获得了对肿瘤细胞生长的显着抑制，而通过常规质粒载体转移相同的基因则产生了显着的抑制肿瘤细胞生长的效果。配备基于 EBV 的质粒载体的非病毒手段提供了适用于免疫基因治疗的新基因操作策略。