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限制， 两个不同但相关的问题：（1）供体器官严重短缺;（2）次优 由于慢性同种异体免疫介导的损伤导致移植物寿命延长。对于终末期肾病患者 疾病，这些限制是显而易见的，在美国有超过90，000人 等待肾脏移植这种严重的供体肾脏短缺是由 移植同种异体移植物的寿命不理想，中位肾移植物存活率仅为8-12 尽管免疫抑制方面取得了进展。这些重大限制表明， 开发新方法以改善供体肾的功能并增强其功能的未满足需求 移植物寿命。用基因疗法治疗供体器官长期以来被认为是 这是一种有希望的增强移植物功能和降低移植物免疫原性的策略， 最近还没有以器官特异性方式进行基因递送的可行方法。 在过去的十年中，离体器官灌注系统的临床开发已经创造了 一个理想的平台，用于选择性地将基因疗法直接递送到供体同种异体移植物。推进 这种临床应用的方法需要在非人类灵长类动物移植模型中进行测试 使用临床相关的免疫抑制方案。为此，我们召集了一个 一组具有离体器官灌注、使用腺相关病毒 (AAV)用于基因治疗、免疫管理和肾移植的载体。We have 3 具体目标：1）优化用于递送基因疗法的离体机器灌注方法 2）确定同种免疫对自体移植模型中肾移植物的影响， 对肾同种异体移植物中转基因表达的应答，和3）开发具有以下功能的新型AAV载体： 人类肾脏移植的趋向性。该项目的成功完成将证明使用 在移植肾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个新的范式，基因增强固体 实体器官移植中的同种异体器官移植和转化途径。

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.