Deveelopment of Novel Gene Theray Strategies Using Recombinant Sendai Virus.

我们最近开发了一种基于仙台病毒(重组仙台病毒：SEV)的新型基因转移载体。本项目的目的是阐明：1)SeV的可能靶器官，2)SeV在体内的生物学行为，以及3)利用小动物进行临床基因治疗的原理研究。与腺病毒载体相比，SeV在呼吸系统中获得了惊人的高基因转移效率(Natural Biotechnol 2000)。在治疗性血管生成方面，表达碱性成纤维细胞生长因子/成纤维细胞生长因子(bFGFR/FGF2)的血管内皮生长因子基因转移在小鼠严重肢体缺血中显示出比以质粒为基础或血管内皮生长因子基因转移为基础的血管内皮生长因子基因转移在CL…中具有更高的治疗潜力更多的美国临床试验(Circ res 2002(1))，这种高疗效依赖于内源性血管内皮生长因子和肝细胞生长因子活性(Circ res 2002(2))。此外，SeV-FGF2不仅能增加慢性肢体缺血的血液灌注量，还能抑制移植静脉的内膜增生(Am J Physiol2003)。在此基础上，提出了SeV-FGF2基因治疗严重肢体缺血患者的临床方案。它已经通过了机构审查委员会，现在正在接受政府审查委员会的评估。基于良好制造规范(GMP)的临床级别的SeV-FGF2已经被制备出来，因此这项临床研究将于2003年开始。此外，我们还展示了SeV在血管壁(FASE B J 2001)、视网膜色素上皮(Exp Eye RES 2002)、活化T淋巴细胞(基因治疗2003，正在出版中)中的高基因转移效率。此外，我们还发现在目前可用的载体中，SeV是最有效的载体，可以将基因转移到CD34阳性的造血干细胞(基因治疗2003)。显然，本项目不仅在基础实验研究方面进行了高质量的工作，而且提出了一种利用SEV进行临床治疗的有希望的策略。我们将把本项目中的这些发现扩展到更可行和更有效的基因治疗。较少

We recently developed a novel gene transfer vector, based on Sendai virus (recombinant Sendai virus: SeV). Aims of the current project were to clarify; 1) possible target organs of SeV, 2) biological behavior of the SeV in vivo, and 3) proof of principle studies for clinical gene therapy using small animals.In respiratory system, SeV achieved amazingly high gene transfer efficiency compared to adenoviral vectors (Nature Biotechnol 2000). Using this system, we demonstrated that SeV-mediated interleukin-10 (IL-10) gene transfer targeted to the airway epithelium efficiently inhibited post transplantation fibrous airway obliteration in mice (Gene Ther 2003).Regarding the application of SeV for therapeutic angiogenesis, SeV expressing basic fibroblast growth factor (bFGF/FGF-2) constantly showed higher therapeutic potentials in murine critical limb ischemia, compared to that seen using plasmid-based or SeV-mediated vascular endothelial growth factor gene transfer, which has been used for cl … More inical trials in United Stales (Circ Res 2002(1)), This high therapeutic effect depended on the endogenous VEGF and hepatocyte growth factor activities (Circ Res 2002 (2)). Further, SeV-FGF2 was also effective not only to increase blood perfusion in chronic limb ischemia but also to limit intimal hyperplasia of vein grafts (Am J Physiol 2003, in press).Based on these findings, clinical protocol of SeV-FGF2 gene therapy to treat patients with critical limb ischemia was proposed. This already passed Institutional Review Board, and is now under evaluation by governmental review board. Clinical grade SeV-FGF2 based on good manufacturing practice (GMP) was already prepared, thus this clinical study will be started in 2003.Further, we demonstrated high gene transfer efficiencies of SeV in vessel wall (FASEB J 2001), retinal pigment epithelium (Exp Eye Res 2002), activated T-lymphocytes (Gene Therapy 2003, in press). In addition, we found that SeV was the most efficient vehicle for gene transfer to CD34 positive hematopoietic stem cells among the currently available vectors (Gene Therapy 2003).Clearly, the current project performed not only high quality works in basic experimental studies, but also suggested a hopeful clinical strategy using SeV.We will extend these findings obtained in this project to more feasible and more effective gene therapy. Less