GENE TRANSFER OF THE CYSTIC FIBROSIS TRANSMEMBRANE REGULATOR

引言：宿主免疫应答和低载体效率 预防有效的囊性纤维化（CF），一种常见的，致命的，遗传性的 疾病CF疾病的肺替代标志物需要大量的 患者数量或研究持续时间长。肺动脉替代疗法 CF基因转移载体的测试是使用上颌窦作为 CF肺病的替代模型。腺相关病毒载体 在I期剂量递增研究中使用AAV-CFTR，随后进行免疫组化。 将CFTR cDNA转移到呼吸道的II期临床试验 CF患者上颌窦上皮细胞。这是 首个CF基因治疗II期研究，测量 CFTR基因转移。 方法：随机、非盲、剂量递增、受试者内， 在上颌窦中进行AAV-CFTR的I期临床试验 10例CF患者的窦口。一项随机、双盲、 安慰剂对照，受试者内II期临床试验正在进行中 入组的22例患者中有16例进展。II期研究的动力 使用22名患者检测鼻窦炎复发率降低50%， 双侧α 0.05和β 0.20。 结果：在I期研究中，最高水平的基因转移是 在活组织检查中在0.1-1 AAV-CFTR载体拷贝/细胞的范围内观察到 治疗后两周。观察到持续时间长达 10周窦跨上皮电位的剂量依赖性变化 对药物干预的反应差异（TEPD）为 在6名受试患者中的5名中观察到。通过RT-PCR测量表达， 在2/2例受试患者中观察到。第二阶段的其他结果尚待确定 完成研究和破盲。两项研究都表明， 很少或没有炎症或免疫反应。 结论：AAV-CFTR给药至上颌窦导致 成功的，剂量依赖性的基因转移到上颌窦， 窦TEPD的改变提示功能影响， 或无细胞病变或宿主免疫应答。需要进一步研究 对于AAV载体，因为它们可以证明可用于CFTR基因转移， 其他体内基因转移疗法。

INTRODUCTION: The host immune response and low vector efficiency have prevented effective cystic fibrosis (CF), a common, fatal, genetic disease. Pulmonary surrogate markers of CF disease require large patient numbers or long study durations. An alternative to pulmonary testing of CF gene transfer vectors is use of the maxillary sinuses as a surrogate model of CF lung disease. An adeno-associated virus vector (AAV-CFTR) was used in a phase I dose-escalation study followed by a phase II clinical trial to transfer CFTR cDNA into respiratory epithelial cells of the maxillary sinus of CF patients. This is the first CF gene therapy phase II study measuring clinical endpoints of CFTR gene transfer. METHODS: A randomized, unblinded, dose-escalation, within-subjects, phase I clinical trial of AAV-CFTR was conducted in the maxillary sinus of 10 CF patients with antrostomies. A randomized, double-blind, placebo controlled, within-subjects phase II clinical trial is in progress with 16 of 22 patients enrolled. The phase II study is powered to detect a 50% decrease in sinusitis recurrence using 22 patients with two-sided alpha 0.05 and beta 0.20. RESULTS: In the phase I study, the highest level of gene transfer was observed in the range of 0.1-1 AAV-CFTR vector copy per cell in biopsies obtained two weeks after treatment. Persistence was observed for up to 10 weeks. Dose-dependent changes in sinus transepithelial potential difference (TEPD) responses to pharmacological intervention were observed in 5 of 6 tested patients. Expression, measured by RT-PCR, was observed in 2 of 2 tested patients. Other phase II results are pending completion of study and breaking of blind. Both studies demonstrated little or no inflammatory or immune responses. CONCLUSION: AAV-CFTR administration to the maxillary sinus results in successful, dose-dependent gene transfer to the maxillary sinus and alterations in sinus TEPD suggestive of a functional effect, with little or no cytopathic or host immune response. Further study is warranted for AAV vectors as they may prove useful for CFTR gene transfer and other in vivo gene transfer therapies.