Genetic engineering of kidney allografts by ex vivo perfusion delivery of adeno-associated viral vectors

摘要 尽管在许多领域取得了进展，但实体器官移植领域仍然受到以下限制 两个截然不同但相互关联的问题：(1)供体器官严重短缺和(2)不理想 慢性同种异体免疫介导的损伤导致的移植物寿命。适用于终末期肾病患者 疾病，这些限制是显而易见的，在美国有90,000多人 等待肾移植。供体肾脏的严重短缺又因 同种异体移植肾的寿命低于最佳，移植肾存活率中位数仅为8-12 几年来，尽管在免疫抑制方面取得了进展。这些显著的限制表明， 未得到满足的需要开发新的方法来改善捐赠者肾脏的功能并增强 嫁接长寿。长期以来，用基因疗法治疗供体器官一直被认为是 一种有望增强移植物功能和降低移植物免疫原性的策略，但直到 最近，还没有以特定器官的方式进行基因传递的可行方法。 在过去的十年中，体外器官灌流系统的临床发展创造了 为选择性地直接向供者同种异体移植物提供基因治疗提供了理想的平台。前进 这种用于临床的方法需要在非人类灵长类移植模型中进行测试。 使用临床相关的免疫抑制方案。对于这项提案，我们已经组建了一个 在体外器官灌流、使用腺相关病毒方面拥有专业知识的研究团队 (AAV)载体用于基因治疗、免疫管理和肾移植。我们有3个 具体目标：1)优化提供基因治疗的体外机器灌流方法 对于自体移植模型中的肾移植，2)确定同种异体免疫的影响 对转基因在移植肾中表达的反应，以及3)进化新的AAV载体 人肾移植的趋向性。这个项目的成功完成将展示它的使用 在肾脏移植中实现持久转基因表达的基因工程方法。 这种方法有可能建立一种新的遗传增强固体 同种异体器官移植及实体器官移植中的转化途径。

Abstract Despite advances in many domains, the field of solid organ transplantation remains limited by two distinct but connected problems: (1) a critical shortage of donor organs and (2) suboptimal graft longevity due to chronic alloimmune-mediated injury. For patients with end-stage renal disease, these limitations are readily apparent, with over 90,000 individuals in the United States awaiting kidney transplantation. This severe shortfall of donor kidneys is compounded by the suboptimal longevity of transplanted allografts, with a median kidney graft survival of only 8-12 years despite advances in immunosuppression. These significant limitations indicate a clear unmet need to develop novel approaches to improve the function of donor kidneys and enhance graft longevity. The treatment of donor organs with gene therapies has long been recognized as a promising strategy to enhance graft function and diminish graft immunogenicity, but until recently there have not been feasible approaches for gene delivery in an organ-specific manner. Over the last decade, the clinical development of ex vivo organ perfusion systems has created an ideal platform for selectively delivering gene therapies directly to donor allografts. Advancing this approach toward clinical use requires testing in a non-human primate transplant model using clinically relevant immunosuppression regimens. For this proposal, we have assembled a team of investigators with expertise in ex vivo organ perfusion, the use of adeno-associated viral (AAV) vectors for gene therapy, immune management, and kidney transplantation. We have 3 specific aims: 1) Optimize ex vivo machine perfusion approaches for delivery of gene therapies to kidney grafts in an auto-transplant model, 2) Determine the impact of the alloimmune response on transgene expression in kidney allografts, and 3) Evolve novel AAV vectors with tropism for human kidney grafts. Successful completion of this project will demonstrate the use of genetic engineering approaches to achieve durable transgene expression in kidney grafts. This approach has the potential to establish a new paradigm of genetically augmented solid organ allografts and transform approaches in solid organ transplantation.