Black Aaron, Vasireddy Vidyullatha, Chung Daniel C, Maguire Albert M, Gaddameedi Rajashekhar, Tolmachova Tania, Seabra Miguel, Bennett Jean
F.M. Kirby Center for Molecular Ophthalmology and Center for Advanced Retinal and Ophthalmic Therapeutics, Scheie Eye Institute, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
J Gene Med. 2014 May-Jun;16(5-6):122-30. doi: 10.1002/jgm.2768.
Choroideremia (CHM) is a slowly progressive X-linked retinal degeneration that results in a loss of photoreceptors, retinal pigment epithelium and choroid. CHM, the gene implicated in choroideremia, encodes Rab escort protein-1 (REP-1), which is involved in the post-translational activation via prenylation of Rab proteins.
We evaluated AAV8.CBA.hCHM, a recombinant adeno-associated virus serotype 8 (rAAV8) vector, which targets retinal cells efficiently, for both therapeutic effect and safety in vitro and in vivo in a murine model. In vitro studies included western blot analyses and prenylation assays. In vivo studies included ophthalmoscopy, pupillometry, histology and immunofluorescence analysis.
Infection with AAV8.CBA.hCHM induced the expression of REP-1 protein in a dose-responsive fashion. Transduction with AAV8.CBA.hCHM reverses the biochemical and pathogenetic defects in CHM both in vitro and in vivo and showed no safety concerns in the in vivo investigations performed in the present study.
AAV8 is a promising vector for human clinical gene therapy trials for choroideremia.
无脉络膜症（CHM）是一种缓慢进展的X连锁视网膜变性疾病，可导致光感受器、视网膜色素上皮和脉络膜丧失。与无脉络膜症相关的CHM基因编码Rab护送蛋白-1（REP-1），其通过Rab蛋白的异戊二烯化参与翻译后激活。
我们评估了AAV8.CBA.hCHM，一种重组腺相关病毒8型（rAAV8）载体，其能有效靶向视网膜细胞，在小鼠模型中进行了体外和体内的治疗效果及安全性评估。体外研究包括蛋白质印迹分析和异戊二烯化测定。体内研究包括检眼镜检查、瞳孔测量、组织学和免疫荧光分析。
用AAV8.CBA.hCHM感染以剂量反应方式诱导REP-1蛋白表达。用AAV8.CBA.hCHM转导在体外和体内均逆转了CHM的生化和致病缺陷，并且在本研究进行的体内研究中未显示出安全问题。
AAV8是用于无脉络膜症人类临床基因治疗试验的一种有前景的载体。
