Repeat dosing of adeno-associated viral vectors

我们的实验室首次成功地将AAV2CFTR基因转移到肺上皮细胞 啮齿动物、非人灵长类动物和人类CF志愿者。我们证明了AAV载体有很好的 作为基因治疗剂的潜力。源于前一个预算期间的研究导致了第一次使用 人类体内的rAAV。许多临床前和临床试验表明，AAV载体可以安全使用。 然而，与我们在灵长类动物中的结果不同，cftr基因在人类中没有表达。 学习。在非人类灵长类动物和人类中，转导都需要大量滴度的 重组AAV2。因此，本预算期的目标是评估新的AAV-CFR血清型 含有更强大的cftr表达启动子。这一努力是成功的，并导致了选择 由强大的鸡β肌动蛋白(CBA)启动子驱动的AAV1-26-264CFTR。总体目标是 更新是为将AAV1-CFTR开发为治疗剂提供关键的下一步。这个 有待检验的假设是开发一种带有CBA启动子的新AAV1-26-264载体血清型 运送到呼吸道将是安全的，并导致重组基因表达水平的提高。三 总体问题将会得到解答。使用伪型AAV1载体会导致增加 从重组载体中进行表达？使用伪型AAV1载体会导致增加 免疫反应？将新的更高滴度的AAV1-CFR载体气雾剂输送到CF 轻度肺部疾病患者会导致广泛的基因转移和CFTR表达吗？ 相关性(请参阅说明)：

Our lab group demonstrated the first successful CFTR gene transfer by AAV2CFTRto pulmonary epithelium of rodents, non-human primates, and human CF volunteers. We showed that AAV vectors have great potential as gene therapeutic agents. Studies originating from a prior budget period lead to the first use of rAAV in humans. Many pre-clinical and clinical trials showed that AAV vectors can be used safely. However, unlike our results in primates, CFTR mRNA expression has not been demonstrated in human studies. In both non-human primates and humans, transduction requires the instillation of large titers of recombinant AAV2. Thus the goal of current budget period was to evaluate new AAV-CFTRserotypes containing more powerful promoters of CFTR expression. This effort was successful and lead to the choice of AAV1-26-264CFTR driven by a powerful chicken beta actin (CBA) promoter. The overall goal the renewal is to provide the critical next steps in developing AAV1 -CFTR as a therapeutic agent. The hypothesis to be tested is that development of a new AAV1-26-264 vector serotype with CBA promoter delivered to the airways will be safe and result in increased levels of recombinant gene expression. Three overall questions will be addressed. Will dosing with a pseudotyped AAV 1 vector leads to increased expression from the recombinant vector? Will dosing with a pseudotyped AAV 1 vector leads to increased immune response? Does aerosol delivery of new higher titer AAV1-CFTRvectors administered to CF patients with Mild Lung Disease lead to widespread gene transfer and CFTR expression? RELEVANCE (See instructions):