Selective gene transfer into the liver of non-human primates with E1-deleted, E2A-defective, or E1-E4 deleted recombinant adenoviruses.
Selective gene transfer into the liver of non-human primates with E1-deleted, E2A-defective, or E1-E4 deleted recombinant adenoviruses.
复制标题
使用 E1 缺失、E2A 缺陷或 E1-E4 缺失的重组腺病毒选择性地将基因转移到非人灵长类动物的肝脏中。
DOI：
10.1089/hum.1998.9.5-671
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Wilson,JM
中科院分区：
文献类型：
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作者：
Raper,SE;Haskal,ZJ;Ye,X;Pugh,C;Furth,EE;Gao,GP;Wilson,JM
临床前研究旨在探讨重组腺病毒转染非人灵长类动物肝动脉的可行性和安全性。所使用的载体是在E1中缺失的重组腺病毒，它们含有E2a基因的温度敏感突变，该突变在不允许的温度下编码有缺陷的dna结合蛋白，或者E4区域的缺失，包括开放阅读框（ORF） 6。6只体重8至10公斤的狒狒接受了股动脉插管，并使用血管造影技术通过右肝动脉分支或肝总动脉选择性地将载体引入肝脏右叶的一部分。分别于第4、29、61天进行尸检。对存活29天和61天的狒狒进行了连续的肝脏活检。在2只载体转导到右肝动脉的狒狒中，肝脏的X-Gal组织化学分析显示，基因在靶叶的转移量增加；然而，基因转移存在于整个肝脏。组织病理学定量分析显示，在最高剂量的载体介导下，整个肝脏都存在门脉炎症。尽管观察到基因表达的定性和定量差异，但在靶向和非靶向叶中门静脉炎症水平没有差异。从靶叶和非靶叶分离的总细胞DNA的Southern blot分析显示，整个肝脏的病毒DNA水平相似。聚合酶链反应（PCR）分析能够检测到高剂量载体处理狒狒生殖腺和脑以及许多其他组织中的病毒DNA序列。在用低剂量表达人鸟氨酸转氨基甲酰基酶（OTC）基因的E1-E4缺失载体处理的狒狒中，在第29天和第61天，通过巢式PCR在肝脏中检测到DNA，但在性腺中检测不到DNA。结果表明，动脉给药重组腺病毒E1-E4缺失载体是可行且安全的。在高剂量的载体下，可以看到载体DNA的广泛传播。在低剂量下，肝脏基因转移与载体DNA传播到性腺无关。
Preclinical studies were designed to investigate the feasibility and safety of recombinant adenoviruses transduced into the hepatic artery of nonhuman primates. The vectors used are recombinant adenoviruses deleted in E1 and contain either a temperature-sensitive mutation in the E2a gene, which encodes a defective DNA-binding protein at nonpermissive temperatures, or a deletion of the E4 region, including open reading frame (ORF) 6. Six 8- to 10-kg baboons underwent femoral artery cannulation, and angiographic techniques were used to introduce vector selectively into either a portion of the right lobe of the liver via a branch of the right hepatic artery or the common hepatic artery. Necropsies were performed at 4, 29, or 61 days. Serial sequential liver biopsies were performed in the baboons that survived 29 or 61 days. In the 2 baboons with vector transduction into the right hepatic artery, X-Gal histochemical analysis of the liver showed evidence of quantitatively increased gene transfer in the targeted lobe; however, gene transfer was present throughout the liver. Quantitative analysis of histopathology showed that portal inflammation was present throughout both livers transduced with the highest dose of vector. No differences were seen in the level of portal inflammation in targeted and untargeted lobes despite the observed qualitative and quantitative differences in gene expression. Southern blot analysis of total cellular DNA isolated from targeted and nontargeted lobes showed similar levels of viral DNA throughout the liver. Polymerase chain reaction (PCR) analysis was able to detect viral DNA sequence in gonads and brain as well as many other tissues in baboons treated with high-dose vector. In baboons treated with lower doses of an E1-E4 deleted vector expressing the human ornithine transcarbamylase (OTC) gene, DNA was detectable by nested PCR in liver but not gonads at days 29 and 61. The data suggest that intraarterial administration of recombinant adenoviral E1-E4 deleted vector is feasible and safe. At high doses of vector, widespread dissemination of vector DNA is seen. At low doses, hepatic gene transfer is not associated with vector DNA dissemination to gonads.