A CONTROLLED-STUDY OF ADENOVIRAL-VECTOR-MEDIATED GENE-TRANSFER IN THE NASAL EPITHELIUM OF PATIENTS WITH CYSTIC-FIBROSIS
A CONTROLLED-STUDY OF ADENOVIRAL-VECTOR-MEDIATED GENE-TRANSFER IN THE NASAL EPITHELIUM OF PATIENTS WITH CYSTIC-FIBROSIS
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DOI：
10.1056/nejm199509283331302
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发表时间：
1995-09-28
影响因子：
158.5
通讯作者：
BOUCHER, RC
中科院分区：
文献类型：
--
作者：
KNOWLES, MR;HOHNEKER, KW;BOUCHER, RC
背景。囊性纤维化是一种单基因疾病，它破坏气道中多个离子运输系统，最终导致慢性感染和肺部破坏。通过基因转移将囊性纤维化跨膜传导调节因子（CFTR）基因的正常拷贝引入气道上皮是纠正囊性纤维化患者潜在缺陷的一种有吸引力的方法。我们用腺病毒载体在这些患者的鼻上皮细胞中测试基因治疗的可行性。采用随机、盲法将含有正常CFTR互补DNA的腺病毒载体以四种对数递增剂量（估计感染多重度，1,10,100和1000）或载体单独施用于12例囊性纤维化患者的鼻上皮。通过检测CFTR信使RNA表达的分子技术和经上皮电位差（pd）的功能测量来定量基因转移，以评估囊性纤维化特异性离子转运的异常。通过鼻灌洗和活检评估炎症和载体复制来监测该治疗的安全性。腺病毒载体在给药后8天内通过培养、聚合酶链反应（PCR）或两种剂量依赖性方式在鼻灌洗液中检测。在接受两种最高剂量治疗的6名患者中，有5名患者通过逆转录酶PCR检测或原位杂交发现了基因转移的分子证据。然而，该载体转染的上皮细胞百分比非常低(
Background. Cystic fibrosis is a monogenic disease that deranges multiple systems of ion transport in the airways, culminating in chronic infection and destruction of the lung. The introduction of a normal copy of the cystic fibrosis transmembrane conductance regulator (CFTR) gene into the airway epithelium through gene transfer is an attractive approach to correcting the underlying defects in patients with cystic fibrosis. We tested the feasibility of gene therapy using adenoviral vectors in the nasal epithelium of such patients.Methods. An adenoviral vector containing the normal CFTR complementary DNA in four logarithmically increasing doses (estimated multiplicity of infection, 1, 10, 100, and 1000), or vehicle alone, was administered in a randomized, blinded fashion to the nasal epithelium of 12 patients with cystic fibrosis. Gene transfer was quantitated by molecular techniques that detected the expression of CFTR messenger RNA and by functional measurements of transepithelial potential differences (PDs) to assess abnormalities of ion transport specific to cystic fibrosis. The safety of this treatment was monitored by nasal lavage and biopsy to assess inflammation and vector replication.Results. The adenoviral Vector was detected in nasal-lavage fluid by culture, the polymerase chain reaction (PCR), or both in a dose-dependent fashion for up to eight days after vector administration. There was molecular evidence of gene transfer by reverse-transcriptase PCR assays or in situ hybridization in five of six patients treated at the two highest doses. However, the percentage of epithelial cells transfected by the vector was very low (