Efficient hepatic delivery and expression from a recombinant adeno-associated virus 8 pseudotyped α1-antitrypsin vector
Efficient hepatic delivery and expression from a recombinant adeno-associated virus 8 pseudotyped α1-antitrypsin vector
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DOI：
10.1016/j.ymthe.2005.05.016
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发表时间：
2005-11-01
影响因子：
12.4
通讯作者：
Flotte, TR
中科院分区：
文献类型：
--
作者：
Conlon, TJ;Cossette, T;Flotte, TR
α 1-抗胰蛋白酶（AAT）缺乏是一种单基因疾病，其中AAT（批准符号SERPINA1）基因（PI*Z）突变导致蛋白质错误折叠，失去AAT对肺部的保护性抗蛋白酶作用，并对肝细胞产生毒性作用。AAT缺乏的最佳治疗方法将需要高百分比的肝细胞转导对肝脏和肺部疾病有效。最近，用血清型7和血清型8的衣壳进行假型的rAAV基因组显示出有效的肝转导。我们假设在门静脉注射到靶肝细胞后，血清型8能更好地转导靶细胞，因此在更大比例和更大数量的细胞中表达hAAT。将携带人AAT转基因的AAV2和1、5、8血清型假型载体以1 × 10（10）粒剂量注射到C57BI/6小鼠体内。在24周的研究中，注射AAV2/8的动物的循环hAAT比AAV2高2倍，比AAV2/1和aav5高3倍。最重要的是，在AAV2/8受试者中，高达40%的肝细胞转基因染色呈阳性，而主要是次要的。因此，伪型AAV8提供了一种载体，可以稳定感染高比例的肝细胞，从而表达治疗性分子来修饰AAT PiZ转录本。
alpha 1-Antitrypsin (AAT) deficiency is a single-gene disorder in which a mutation in the AAT (approved symbol SERPINA1) gene (PI*Z) leads to misfolding of the protein, loss of the protective antiprotease effect of AAT for the lungs, and a toxic effect on hepatocytes. Optimal therapy for AAT deficiency will require a high percentage of hepatocyte transduction to be effective for liver and lung disease. Recently, rAAV genomes pseudotyped with capsids from serotypes 7 and 8 showed efficient hepatic transduction. We hypothesized that upon portal vein injection to target hepatocytes, serotype 8 would better transduce target cells and therefore express hAAT in both a greater percentage of cells and greater amounts. AAV2 and pseudotyped vectors for serotypes 1, 5, and 8 carrying the human AAT transgene were injected at 1 x 10(10) particle doses into C57BI/6 mice. Circulating hAAT from AAV2/8-injected animals showed a 2-log advantage over AAV2 and 3-log increase over AAV2/1 and 5 for the 24-week study. Most significantly, up to 40% of total liver cells stained positive for the transgene in AAV2/8 subjects while remaining primarily episomal. Therefore, pseudotyped AAV8 provides a vehicle to infect a high percentage of hepatocytes stably and thereby express therapeutic molecules to modify AAT PiZ transcripts.