INVIVO TRANSFER OF THE HUMAN CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR GENE TO THE AIRWAY EPITHELIUM
INVIVO TRANSFER OF THE HUMAN CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR GENE TO THE AIRWAY EPITHELIUM
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DOI：
10.1016/0092-8674(92)90213-v
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发表时间：
1992-01-10
期刊：
影响因子：
64.5
通讯作者：
CRYSTAL, RG
中科院分区：
文献类型：
--
作者：
ROSENFELD, MA;YOSHIMURA, K;CRYSTAL, RG
使用含有正常人囊性纤维化（CF）跨膜传导调节因子（CFTR）cDNA的复制缺陷型重组腺病毒（Ad）载体（Ad-CFTR）直接转移到气道上皮进行评估。在体外Ad-CFTR感染的CFPAC-1 CF上皮细胞表达人CFTR mRNA和蛋白质，并证明有缺陷的cAMP介导的Cl-渗透性的纠正。在棉鼠体内肺内引入Ad-CFTR后两天，原位分析表明人CFTR基因在肺上皮中表达。逆转录的肺RNA的PCR扩增证明人CFTR转录物来源于Ad-CFTR，肺RNA的北方分析显示人CFTR转录物长达6周。感染后11-14天，使用抗人CFTR抗体在上皮细胞中检测人CFTR蛋白。虽然安全性和有效性仍有待证明，但这些观察结果表明，体内CFTR基因转移作为CF肺部表现的治疗方法是可行的。
Direct transfer of the normal cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene to airway epithelium was evaluated using a replication-deficient recombinant adenovirus (Ad) vector containing normal human CFTR cDNA (Ad-CFTR). In vitro Ad-CFTR-infected CFPAC-1 CF epithelial cells expressed human CFTR mRNA and protein and demonstrated correction of defective cAMP-mediated Cl-permeability. Two days after in vivo intratracheal introduction of Ad-CFTR in cotton rats, in situ analysis demonstrated human CFTR gene expression in lung epithelium. PCR amplification of reverse transcribed lung RNA demonstrated human CFTR transcripts derived from Ad-CFTR, and Northern analysis of lung RNA revealed human CFTR transcripts for up to 6 weeks. Human CFTR protein was detected in epithelial cells using anti-human CFTR antibody 11-14 days after infection. While the safety and effectiveness remain to be demonstrated, these observations suggest the feasibility of in vivo CFTR gene transfer as therapy for the pulmonary manifestations of CF.