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

通过腺相关病毒载体的离体灌注递送同种异体肾的基因工程

基本信息

批准号：
10667569
负责人：
Aravind Asokan
金额：
$ 47.71万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-19 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10667569
关键词：
Address Adenovirus Vector Allografting Autologous Transplantation Cells Chronic Clinical Cryopreservation Development End stage renal failure Engineering Evolution Family suidae Gene Delivery Gene Transduction Agent Genes Genetic Engineering Goals Graft Survival Green Fluorescent Proteins Heart Transplantation Human Immune Immune response Immunologics Immunosuppression Individual Injury Kidney Kidney Transplantation Libraries Library Associations Longevity Lung Transplantation Mediating Methods Modeling Organ Organ Donor Organ Transplantation Outcome Patients Perfusion Pre-Clinical Model Process Rattus Regimen Reporter Reporter Genes Research Research Personnel Safety Solid System Testing Time Transgenes Transplantation Tropism United States Viral Viral Vector Virus Work adeno-associated viral vector allotransplant body system cell type clinical development clinical translation clinically relevant cross reactivity design ex vivo perfusion experimental study functional improvement gene therapy graft function heart allograft high risk immunogenicity immunoregulation improved kidney allograft liver transplantation nonhuman primate novel novel strategies organ repair overexpression post-transplant programmed cell death ligand 1 rational design response single-cell RNA sequencing targeted delivery therapeutic gene transgene delivery transgene expression transplant model uptake

项目摘要

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.

抽象的尽管在许多领域取得了进展，实体器官移植领域仍然受到以下因素的限制：两个截然不同但又相互关联的问题：(1) 供体器官严重短缺；(2) 次优由于慢性同种免疫介导的损伤而延长移植物的寿命。对于终末期肾病患者疾病，这些限制是显而易见的，美国有超过 90,000 人等待肾移植。供肾严重短缺的情况更加严重移植同种异体移植物的寿命不佳，肾移植物的中位存活率仅为 8-12 尽管免疫抑制方面取得了进展，但仍持续了数年。这些重大限制表明了一个明确的开发新方法来改善供体肾脏功能并增强功能的需求尚未得到满足嫁接长寿。用基因疗法治疗供体器官长期以来一直被认为是增强移植物功能和降低移植物免疫原性的有前途的策略，但直到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发展已经创造了一个有选择地直接向供体同种异体移植物提供基因治疗的理想平台。前进这种临床应用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。对于这个提案，我们准备了一份具有离体器官灌注、腺相关病毒使用方面专业知识的研究人员团队 (AAV) 载体用于基因治疗、免疫管理和肾移植。我们有 3 具体目标：1）优化用于基因治疗的离体机器灌注方法对自体移植模型中的肾移植物，2) 确定同种免疫的影响对肾同种异体移植物中转基因表达的反应，以及 3) 进化新的 AAV 载体对人类肾移植物的趋向性。该项目的成功完成将展示其用途基因工程方法在肾移植物中实现持久的转基因表达。这种方法有可能建立基因增强固体的新范式实体器官移植中的器官同种异体移植和转化方法。