CF Gene Therapy with adeno-associated viral vectors

 描述：CF是一种常染色体疾病，导致患者的发病率和死亡率显着的疾病。我们的实验室进行的实验导致了第一个使用AAV载体的基因治疗试验。目前的提案旨在利用我们过去的专业知识，成功开发一种基于血清1型的新AAV载体，并在CF患者的临床试验中利用这种载体。由于CF新疗法的开发将带来显著受益，因此该申请具有高度相关性。我们已经确定了三种新方法来克服AAV基因治疗CF的挑战：使用AAV 1，其对肺更具营养性;使用27-264，CFTR的截短版本，其通过新的机制纠正CFNF 508;以及包含强大的鸡β-肌动蛋白（CBA）启动子。本申请的总体目标是提供开发AAV 1-CFTR作为治疗剂的关键后续步骤。我们提出了三个具体目标：1。评价AAV 1-B27 -264- CFTR的一般安全性、毒理学、生物分布和免疫应答。2.为了确定在人类临床试验中给予含有截短的CFTR的AAV 1载体是否会导致表达增加。3.评估向灵长类动物重复给予AAV 1-CFTR载体是否导致广泛的基因转移和CFTR表达。新奇：这项工作是新颖的，因为它结合了通过反式互补和基因疗法修复内源性ΔF508 CFTR。

 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.