GENE THERAPY FOR CYSTIC FIBROSIS USING AAV VECTORS

使用 AAV 载体治疗囊性纤维化

• 批准号: 6668336
• 负责人: Arthur Dusty Miller
• 金额: $ 25.65万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6668336
• 关键词: adeno associated virus group; biotechnology; chloride channels; clinical research; clinical trial phase I; cooperative study; cystic fibrosis; gene therapy; genetic markers; genetic transduction; human subject; human therapy evaluation; laboratory mouse; respiratory epithelium; transfection /expression vector

Gene Therapy for Cystic Fibrosis using AAV Vectors. Cystic fibrosis (CF) affects 1 in 3,200 births and leads to debilitating lung disease and premature death at a median age of 32 years. Gene therapy may provide a cure for this disease by replacement of the defective protein, the cystic fibrosis transmembrane regulator (CFTR). The long-range objective of this application is the development of lung-targeted gene therapy for treatment of CF using viral vectors derived from a non-pathogenic human parvovirus, adeno-associated virus (AAV). AAV vectors have been shown to promote gene transfer and long-term expression in several animal models, in particular, AAV vectors can transduce airway and alveolar epithelial cells in the lungs of mice at rates of greater than or equal to 5% and lasting for over 8 months. However, clinical trials involving AAV vector-mediated transfer of the CFTR gene to humans have yet to show useful levels of CFTR expression or clinical efficacy. This lack of expression is likely due to the difficulty of making AAV vectors that express CFTR because of the large size of the CFTR cDNA, and the difficulty of measuring small changes in CF disease severity that might result from gene therapy. To circumvent these difficulties and to provide a more direct test of the utility of AAV vectors in humans, the specific aims of this proposal include clinical trials in healthy subjects and CF patients which will address the safety, efficacy, and immune responses to nasal and bronchial administration of AAV vectors that encode an easily detected histochemical marker gene, human placental alkaline phosphatase (hpAp). We have found that vectors derived from AAV serotype 6 show improved transduction rates in mouse airway compared to commonly used AAV serotype 2 vectors, although more extensive safety data in humans is available for AAV2. Here we propose to compare expression of hpAP extensive safety data in humans is available for AAV2. Here we propose to compare expression of hpAP delivered by both AAV2 and AAV6 in healthy subjects. The vector serotype with the better safety and gene expression profile will be tested subsequently in patients with CF. Another specific aim will address the development of effective AAV vectors for transfer and efficient expression of the CFTR cDNA. These approaches are designed to most efficiently test and develop AAV vectors for treatment of CF.

使用AAV载体的囊性纤维化的基因治疗。囊性纤维化（CF）影响3，200名新生儿中的1名，并导致衰弱性肺病和过早死亡，中位年龄为32岁。基因治疗可以通过替换有缺陷的蛋白质，囊性纤维化跨膜调节因子（CFTR）来治愈这种疾病。本申请的长期目标是开发用于治疗CF的肺靶向基因疗法，所述肺靶向基因疗法使用源自非致病性人细小病毒腺相关病毒（AAV）的病毒载体。AAV载体已经显示在几种动物模型中促进基因转移和长期表达，特别地，AAV载体可以以大于或等于5%的比率感染小鼠肺中的气道和肺泡上皮细胞，并且持续超过8个月。然而，涉及腺相关病毒载体介导的CFTR基因向人类转移的临床试验尚未显示有用的CFTR表达水平或临床功效。这种表达的缺乏可能是由于由于CFTR cDNA的大尺寸而难以制备表达CFTR的AAV载体，以及难以测量可能由基因治疗引起的CF疾病严重程度的微小变化。为了规避这些困难并提供AAV载体在人类中的效用的更直接的测试，该提议的具体目的包括在健康受试者和CF患者中的临床试验，其将解决对编码容易检测的组织化学标记基因人胎盘碱性磷酸酶（hpAp）的AAV载体的鼻和支气管施用的安全性、功效和免疫应答。我们已经发现，与常用的AAV血清型2载体相比，源自AAV血清型6的载体在小鼠气道中显示出改善的转导速率，尽管可获得关于AAV 2的更广泛的人类安全性数据。在这里，我们提出比较hpAP在人类中的表达，广泛的安全性数据可用于AAV2。在这里，我们提出比较健康受试者中由AAV2和AAV6两者递送的hpAP的表达。具有更好安全性和基因表达谱的载体血清型将随后在CF患者中进行测试。另一个具体目标将致力于开发用于转移和有效表达CFTR cDNA的有效AAV载体。这些方法被设计为最有效地测试和开发用于治疗CF的AAV载体。