Division of Cardiothoracic Surgery, Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA.
Division of Cardiothoracic Surgery, Department of Surgery, Columbia University Medical Center, New York City, New York, USA.
Hum Gene Ther. 2023 Apr;34(7-8):303-313. doi: 10.1089/hum.2022.241.
Transplantation, the gold standard intervention for organ failure, is a clinical field that is ripe for applications of gene therapy. One of the major challenges in applying gene therapy to this field is the need for a method that achieves consistent and robust gene delivery to allografts. Normothermic perfusion is a growing organ preservation method and a device for cardiac preservation was recently approved by the Food and Drug Administration (FDA) (Organ Care System, OCS™; TransMedics, Inc., Andover, MA); this device maintains donor hearts in a near physiologic state while they are transported from the donor to the recipient. This study describes the administration of recombinant adeno-associated viral vectors (rAAVs) during normothermic perfusion for the delivery of transgenes to porcine cardiac allografts. We utilized a myocardial-enhanced AAV3b variant, SASTG, assessing its transduction efficiency in the OCS perfusate relative to other AAV serotypes. We describe the use of normothermic perfusion to deliver SASTG carrying the Firefly Luciferase transgene to porcine donor hearts in four heterotopic transplant procedures. Durable and dose-dependent transgene expression was achieved in the allografts in 30 days, with no evidence of off-target transgene expression. This study demonstrates the feasibility and efficiency of delivering genes to a large animal allograft utilizing AAV vectors during perfusion. These findings support the idea of gene therapy interventions to enhance transplantation outcomes.
移植是治疗器官衰竭的金标准干预手段，在该领域，基因治疗的应用已经成熟。将基因治疗应用于该领域的主要挑战之一是需要一种方法，能够将基因稳定且有效地递送到同种异体移植物中。常温灌注是一种新兴的器官保存方法，最近一种心脏保存设备已获得美国食品和药物管理局（FDA）的批准（器官保存系统，OCS™；TransMedics，Inc.，马萨诸塞州安多弗）；该设备可使供体心脏在从供体运输到受体的过程中保持近乎生理状态。本研究描述了在常温灌注期间给予重组腺相关病毒载体（rAAV），以将转基因递送到猪心脏同种异体移植物中。我们利用心肌增强型 AAV3b 变体 SASTG，评估了其在 OCS 灌注液中的转导效率相对于其他 AAV 血清型。我们描述了使用常温灌注将携带萤火虫荧光素酶转基因的 SASTG 递送到四个异位移植程序中的猪供体心脏。在 30 天内，同种异体移植物中实现了持久且剂量依赖性的转基因表达，没有证据表明脱靶转基因表达。本研究证明了在灌注过程中使用 AAV 载体向大型动物同种异体移植物递送基因的可行性和效率。这些发现支持了通过基因治疗干预增强移植结果的想法。
