Inverse zonation of hepatocyte transduction with MV vectors between mice and non-human primates
Inverse zonation of hepatocyte transduction with MV vectors between mice and non-human primates
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DOI：
10.1016/j.ymgme.2011.06.002
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发表时间：
2011-11-01
影响因子：
3.8
通讯作者：
Wilson, James M.
中科院分区：
文献类型：
--
作者：
Bell, Peter;Wang, Lili;Wilson, James M.
以腺相关病毒8型(AAV8)为基础的基因转移载体具有高效的肝脏转导作用，且易于静脉注射。在小鼠中，AAV8主要转导中心静脉附近的肝细胞，并在门静脉周围区域产生较低的转导水平。这种转导偏向对于旨在纠正代谢性肝酶的基因治疗具有重要意义，因为沿着门中轴的代谢区带要求治疗性蛋白在它们通常所在的区段内表达。在本研究中，我们比较了AAV8表达绿色荧光蛋白(GFP)在小鼠、狗和非人灵长类动物肝脏中的表达模式。我们证实了当使用肝脏特异性甲状腺激素结合球蛋白(TBG)启动子时，AAV8转基因小鼠的转基因表达在中心周围占优势，但也观察到了与普遍存在的鸡β-肌动蛋白(CB)和巨细胞病毒(CMV)启动子相同的表达模式，这表明转导区带不是由启动子特异性引起的。在注射AAV8的狗身上也发现了以中心周围为主的表达。相比之下，食蟹猴和恒河猴AAV载体的表达模式相反，即转基因在门静脉区域表达最强，在中心静脉周围表达较弱或缺失。然而，注射AAV8的幼年恒河猴和新生小鼠显示出转基因表达的随机分布，既没有门脉也没有中心转导偏向。根据猴子的数据，经AAV载体治疗的成年人也可能主要在门脉周围区域表达转基因，而婴儿可能在肝脏中表现出统一的转导模式。(C)2011 Elsevier Inc.保留所有权利。
Gene transfer vectors based on adeno-associated virus 8 (AAV8) are highly efficient in liver transduction and can be easily administered by intravenous injection. In mice, AAV8 transduces predominantly hepatocytes near central veins and yields lower transduction levels in hepatocytes in periportal regions. This transduction bias has important implications for gene therapy that aims to correct metabolic liver enzymes because metabolic zonation along the porto-central axis requires the expression of therapeutic proteins within the zone where they are normally localized.In the present study we compared the expression pattern of AAV8 expressing green fluorescent protein (GFP) in liver between mice, dogs, and non-human primates. We confirmed the pericentral dominance in transgene expression in mice with AAV8 when the liver-specific thyroid hormone-binding globulin (TBG) promoter was used but also observed the same expression pattern with the ubiquitous chicken beta-actin (CB) and cytomegalovirus (CMV) promoters, suggesting that transduction zonation is not caused by promoter specificity. Predominantly pericentral expression was also found in dogs injected with AAV8. In contrast, in cynomolgus and rhesus macaques the expression pattern from AAV vectors was reversed, i.e. transgene expression was most intense around portal areas and less intense or absent around central veins. Infant rhesus macaques as well as newborn mice injected with AAV8 however showed a random distribution of transgene expression with neither portal nor central transduction bias. Based on the data in monkeys, adult humans treated with AAV vectors are predicted to also express transgenes predominantly in periportal regions whereas infants are likely to show a uniform transduction pattern in liver. (C) 2011 Elsevier Inc. All rights reserved.