GENE TRANSFER OF THE CYSTIC FIBROSIS TRANSMEMBRANE REGULATOR

宿主免疫反应和低载体效率是关键障碍。 有效的囊性纤维化跨膜电导调节剂(CFTR) 分别用腺病毒载体和脂质体载体进行基因转移。腺体- 相关病毒介导的CFTR转导(AAV-CTR)是 对CF患者有吸引力的潜在基因治疗剂，因为 在动物研究中长期表达和低致病性。AAV-CFTR 在一项I期剂量递增研究中被使用，以将CFTRcDNA转移到 10例慢性阻塞性肺疾病患者上颌窦呼吸道上皮细胞 上颌窦是评价CF新疗法的有吸引力的对象 因为它们有相同的离子传输系统和微生物学 下呼吸道，并允许局部递送已知 药物浓度直接作用于人呼吸道上皮细胞。 半定量聚合酶链式反应检测剂量依赖性基因转移 观察剂量大于1 X 10e4复制单位(RU)AAV- CFTR。在1×10e5 RU AAV-CFTR上，观察到了基因转移 在两周后获得的活检组织中每个细胞0.1-1个载体拷贝 治疗。持续观察长达10周。功能性 窦性跨上皮电位差(TEPD)的恢复 被观察到了。观察到剂量依赖的超极化。 对低氯和异丙肾上腺素的反应。5个10e4和1个10e5 RU AAV-CFTR，超极化6.9(+/-)3.7和10.8(+/-)4.7 mV (平均(+/-)S.D.)。与之形成鲜明对比的是 腺病毒载体，很少或没有免疫反应或炎症 观察，即使在重复给药后也是如此。这一阶段研究的结果是 提示AAV-CFTR应用于上颌窦可导致 成功地将剂量依赖的基因转移到上颌窦 鼻窦TEFP部分纠正，宿主免疫反应很少或无反应。 基于这些结果，II期，双盲，安慰剂对照， 1×10~(-5)RU剂量AAV-CFTR作用的受试者内研究 对鼻窦炎的临床复发进行了计划。这表示 首次有机会研究CFTR基因转移对A细胞的影响 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.