CF Gene Therapy with adeno-associated viral vectors

 描述：CF是一种常染色体疾病，在这种疾病的患者中会导致显著的发病率和死亡率。我们实验室进行的实验导致了使用AAV载体进行的第一次基因治疗试验。目前的建议是利用我们过去的专业知识，成功地开发出一种基于1型血清的新的AAV载体，并将该载体用于CF患者的临床试验。由于CF新疗法的开发将带来显著的好处，因此这一应用具有很高的相关性。我们已经确定了三种新的方法来克服针对CF的AAV基因治疗的挑战：使用对肺更有营养的AAV1；使用27-264，通过一种新的机制纠正F508的CFTR27-264版本；以及包括强大的鸡β-肌动蛋白(Cba)启动子。本申请的总体目标是为AAV1-CFTR作为治疗剂的开发提供关键的下一步。我们提出了三个总体的具体目标：1.评价AAV1-的总体安全性、毒理学、生物分布和免疫应答。2.确定在人类临床试验中，使用包含截短cftr的AAV1载体是否会导致表达增加。3.评估AAV1-CFTR载体在灵长类动物体内的重复给药是否导致广泛的基因转移和CFTR的表达。新颖性：这项工作具有新颖性，因为它结合了内源性ΔF508CFTR的反互补修复和基因治疗。

 DESCRIPTION: CF is an autosomal disease that leads to significant morbidity and mortality in patients with the disorder. Experiments conducted by our laboratories led to the first gene therapy trial using AAV vectors. The current proposal is designed to utilize our past expertise to successfully develop a new AAV vector based upon serotype 1 and to utilize this vector in clinical trials in CF patients. Because of the significant benefit that would be derived from the development of new therapies for CF, this application is highly relevant. We have identified three new approaches to overcome challenges to AAV gene therapy for CF: use of AAV1, which is more trophic for the lung; use of 27-264, a truncated version of CFTR that corrects F508 by a novel mechanism; and inclusion of a powerful chicken β-actin (CBA) promoter. The overall goal of this application is to provide the critical next steps in developing AAV1-CFTR as a therapeutic agent. We propose three overall Specific Aims: 1. To evaluate the general safety, toxicology, biodistribution, and immune response to AAV1-27-264- CFTR. 2. To determine whether dosing with an AAV1 vector containing a truncated CFTR will lead to increased expression in a human clinical trial. 3. To assess whether repeat dosing of AAV1-CFTR vectors administered to primates leads to wide- spread gene transfer and CFTR expression. Novelty: This work is novel because it combines the repair of endogenous ΔF508 CFTR by transcomplementation and gene therapies.