Engelhardt J F, Simon R H, Yang Y, Zepeda M, Weber-Pendleton S, Doranz B, Grossman M, Wilson J M
Institute for Human Gene Therapy, University of Pennsylvania Medical Center, Wistar Institute, Philadelphia 19104.
Hum Gene Ther. 1993 Dec;4(6):759-69. doi: 10.1089/hum.1993.4.6-759.
We have evaluated the biological efficacy of E1-deleted adenoviruses in baboons for lung-directed gene therapy of cystic fibrosis (CF). The experimental design attempted to simulate a phase I clinical trial with animals receiving a single dose of virus to an isolated pulmonary segment. A total of 14 animals divided into four groups, each of which received escalating doses of virus, were used. Individual animals were necropsied 4 and 21 days after gene transfer and tissues were carefully surveyed for gene expression. Expression of the transgene was localized primarily to the area into which it was infused; the efficiency of recombinant gene expression and the abundance of transgene sequences were proportional to dose and both diminished with time. Transgene expression was found predominantly in alveolar cells with patches of expression in the proximal and distal airway. Analysis of adenoviral protein expression within transgene-expressing cells revealed infrequent expression of the E2a gene and no detectable expression of late genes (i.e., fiber protein). These results suggest that recombinant adenovirus can be used to transfer genes efficiently to the lung of nonhuman primates and that therapeutic strategies of cystic fibrosis may require repetitive administration with current vectors.
我们评估了E1缺失腺病毒在狒狒中用于囊性纤维化（CF）肺定向基因治疗的生物学疗效。实验设计试图模拟I期临床试验，让动物接受单剂量病毒注入孤立的肺段。总共使用了14只动物，分为四组，每组接受递增剂量的病毒。在基因转移后4天和21天对个体动物进行尸检，并仔细检查组织中的基因表达。转基因的表达主要定位于注入区域；重组基因表达效率和转基因序列丰度与剂量成正比，且均随时间降低。转基因表达主要见于肺泡细胞，在近端和远端气道有散在表达。对转基因表达细胞内腺病毒蛋白表达的分析显示，E2a基因表达罕见，晚期基因（即纤维蛋白）无可检测表达。这些结果表明，重组腺病毒可有效用于将基因转移至非人灵长类动物肺中，并且囊性纤维化的治疗策略可能需要使用当前载体进行重复给药。
