GENE TRANSFER OF THE CYSTIC FIBROSIS TRANSMEMBRANE REGULATOR

宿主免疫反应和载体效率低是主要障碍 有效的囊性纤维化跨膜传导调节因子（CFTR） 分别通过腺病毒和脂质体载体进行基因转移。 腺- 相关病毒介导的CFTR转导（AAV-CTR）是一种新的免疫抑制剂。 对CF患者有吸引力的潜在基因治疗剂， 在动物研究中的长期表达和低致病性。 AAV-CFTR 用于I期剂量递增研究，以将CFTR cDNA转移到 10例CF患者上颌窦呼吸道上皮细胞。 上颌窦是有吸引力的评价新的治疗CF 因为它们有着和人类一样的离子传输系统和微生物学 下呼吸道，并允许已知的局部递送 将药物浓度直接施用至人呼吸道上皮细胞。 通过半定量PCR检测剂量依赖性基因转移， 在大于IX IOe 4个复制单位（RU）AAV的剂量下观察到。 CFTR。 在1 X 10 e5 RU AAV-CFTR下，观察到基因转移范围为 0.1-1个载体拷贝/细胞， 治疗 观察到持续长达10周。 功能 窦跨上皮电位差（TEPD）的恢复 观察了 观察到剂量依赖性超极化， 对低氯化物和异丙肾上腺素的反应。 5 X 10 e4和1 X10 e5 RU AAV-CFTR，超极化为6.9（+/-）3.7和10.8（+/-）4.7 mV (mean（+/-）标准差，分别观察到。 相比 腺病毒载体，很少或没有免疫反应或炎症， 即使在重复给药后也观察到。 I期研究的结果 这表明AAV-CFTR施用于上颌窦导致 成功的，剂量依赖性的基因转移到上颌窦， 窦TEFP的部分校正和很少或没有宿主免疫应答。 基于这些结果，一项II期、双盲、安慰剂对照， 在1 X 10 e5 RU剂量下对AAV-CFTR的作用的受试者内研究 对鼻窦炎的临床复发进行研究。 这代表 第一次有机会研究CFTR基因转移对 临床终点

The host immune response and low vector efficiency are key impediments to effective cystic fibrosis transmembrane conductance regulator (CFTR) gene transfer by adenoviral and liposomal vectors, respectively. Adeno- associated virus mediated transduction of CFTR (AAV-CTR) is an attractive potential gene therapeutic agent for CF patients because of long-term expression and low pathogenicity in animal studies. AAV-CFTR was used in a phase I dose-escalation study to transfer CFTR cDNA into respiratory epithelial cells in the maxillary sinus of 10 CF patients. The maxillary sinuses are attractive for evaluating new treatments of CF because they have the same ion transport systems and microbiology as the lower respiratory tract and allow localized delivery of known concentrations of agents directly to human respiratory epithelial cells. Dose dependent gene transfer, assayed by semi-quantitative PCR, was observed at doses of greater that 1 X 10e4 replication units (RU) AAV- CFTR. At 1 X 10e5 RU AAV-CFTR, gene transfer was observed in the range of 0.1-1 vector copy per cell in biopsies obtained two weeks after treatment. Persistence was observed for up to 10 weeks. Functional restoration of the sinus transepithelial potential differences (TEPD) was observed. Dose-dependent hyperpolarization was observed in response to low chloride and isoproterenol. At 5 X 10e4 and 1 X10e5 RU AAV-CFTR, hyperpolarizations of 6.9 (+/-) 3.7 and 10.8 (+/-) 4.7 mV (mean (+/-) s.d., respectively, were observed. In contrast to adenoviral vectors, little or no immune response or inflammation was observed, even after repeat dosing. The results of this phase I study suggest that AAV-CFTR administration to the maxillary sinus results in successful, dose-dependent gene transfer to the maxillary sinus with partial correction of sinus TEFP and little or no host immune response. Based on these results, a phase II, double-blind, placebo-controlled, within-subjects study on the effect of AAV-CFTR at a dose of 1 X 10e5 RU on the clinical recurrence of sinusitis is planned. This represents the first opportunity to study the effects of CFTR gene transfer on a clinical endpoint.