Repeat Dosing of Adeno-Associated Viral Vectors

腺相关病毒载体的重复给药

• 批准号: 6853341
• 负责人: William B. Guggino
• 金额: $ 29.16万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6853341
• 关键词: Macaca mulatta; adeno associated virus group; aerosols; biotechnology; bronchoscopy; capsid; chloride channels; clinical trials; cystic fibrosis; dosage; gene delivery system; gene expression; gene therapy; human subject; human therapy evaluation; immune response; inflammation; inhalation drug administration; lung disorder; patient oriented research; recombinant virus; respiratory epithelium; transfection; transfection /expression vector; virus antigen; virus protein

In animal and in the first human studies, our group showed that AAV2-CFTR could be applied to the nasal epithelium, maxillary sinus, and right lower lobe of the human lung without any adverse affects. These studies and multiple animal studies published in the literature showed that genes expressed from AAV vectors result in persistent expression in a variety of tissues. A recent multi institutional collaborative study showed statistically significant improvement in FEV1 at Day 30 and induced sputum IL-8 levels observed between Days 14-45 following 3 aerosolized doses of 1 x 10[13] DNAse resistant particles tgAAVCF. Thus, AAV vectors have great potential as therapeutic agents. Despite these promising studies several hurdles must be overcome before AAV vectors will be useful therapeutic agents. This grant will use a combination of animal and human studies to address the hypothesis that development of a new serotype with new more powerful promoters delivered to the airways efficiently via a bronchoscopic Microsprayer will be safe and result in increased levels of recombinant gene expression. There are three overall questions that will be addressed. 1. Will Dosing with a pseudotyped vector based on the AAV5 virus lead to increased expression from the recombinant vector? Experiments in this specific aim will be directed specifically toward addressing, if delivery of AAV5-CFTR to the mammalian airway elicits an immunologic or inflammatory response and if so to what aspect of the vector is the immunologic response directed. 2. Will inclusion of a more powerful promoter augment CFTR expression from recombinant vectors? Thus, we will explore the potential risks and benefits of using an altered CFTR construct and a new chicken beta-actin promoter to enhance expression of CFTR. It is anticipated that answers to these questions in a non-human primate model will provide the preclinical data necessary to launch a study of aerosolized AAV5-CFTR vector in patients with CF. 3. Does aerosol delivery of new higher titer AAV5-CFTR vectors administered to CF patients with Mild Lung Disease lead to uniform gene transfer and CFTR expression? Particular attention will also be given to whether the new pseudotyped vector AAV5-CFTR in humans leads to increased expression of the recombinant vector. It is expected that answers to this question will lead to a practical delivery system for recombinant AAV5-CFTR with potential new promoters to boost CFTR expression.

在动物和第一个人类研究中，我们的小组表明，AAV2-CFTR可以应用于人类肺的鼻上皮，上颌窦和右下叶，而不会产生任何不利影响。这些研究和文献中发表的多项动物研究表明，从AAV载体表达的基因导致各种组织中持续表达。最近的一项多机构合作研究显示，在第30天，FEV1的统计学显着改善，并在3天之间在第14-45天之间观察到了痰液IL-8水平，3 3例1 x 10 [13] DNase抗性颗粒TGAAVCF。因此，AAV载体作为治疗剂具有巨大的潜力。尽管有这些有前途的研究，在AAV载体是有用的治疗剂之前，必须克服几个障碍。这笔赠款将使用动物的组合 人类的研究是为了解决以下假设，即通过支气管镜微喷雾剂有效地传递新的更强大的启动子的新血清型将是安全的，并导致重组基因表达的水平增加。将解决三个总体问题。 1。基于AAV5病毒的伪型载体的给药会导致重组载体的表达增加吗？如果将AAV5-CFTR递送到哺乳动物气道中会引起免疫学或炎症反应，并且如果是这样，则针对载体或炎症反应的实验将专门针对解决方案。 2。是否将重组向量的更强大的启动子增强CFTR表达？因此，我们将探索潜在的风险 使用改变的CFTR构建体和新的鸡β-肌动蛋白启动子来增强CFTR的表达的好处。可以预计，在非人类灵长类动物模型中对这些问题的答案将提供临床前数据，以启动CF患者的雾化AAV5-CFTR载体的研究。 3。向患有轻度肺部疾病的CF患者施用的新高滴度AAV5-CFTR载体的气溶胶是否会导致基因转移和CFTR表达均匀？还将特别注意人类中新的伪型载体AAV5-CFTR是否导致重组载体的表达增加。可以预期，这个问题的答案将导致重组AAV5-CFTR具有潜在新启动子以增强CFTR表达的实用输送系统。