Natkunarajah M, Trittibach P, McIntosh J, Duran Y, Barker S E, Smith A J, Nathwani A C, Ali R R
Division of Molecular Therapy, UCL Institute of Ophthalmology, University College London, London, UK.
Gene Ther. 2008 Mar;15(6):463-7. doi: 10.1038/sj.gt.3303074. Epub 2007 Nov 15.
To date adeno-associated viral (AAV) vectors are the only gene therapy vectors that have been shown to efficiently transduce photoreceptor cells and have thus become the most commonly used vector for ocular transduction. Various AAV serotypes have been evaluated in the eye, the first of which was AAV2, which is able to transduce photoreceptors, retinal pigment epithelium (RPE) and retinal ganglion cells. AAV serotypes 1 and 4, as well as AAV2 pseudotyped with these capsids, only transduce the RPE. AAV serotype 5 and AAV2/5 transduce the photoreceptors as well as RPE, but not retinal ganglion cells. Here, we assessed the capacity of the novel serotype AAV2/8 to transduce various ocular tissues of the adult murine retina by administering AAV2/8 green fluorescent protein intravitreally, subretinally and intracamerally. We also determined the kinetics and efficiency of self-complementary AAV (scAAV) vectors of serotypes 2/2, 2/5 and 2/8 and compared them with single-stranded AAV (ssAAV). We found that ssAAV2/8 transduces photoreceptors and RPE more efficiently than ssAAV2/2 and ssAAV2/5, and that scAAV2/8 had faster onset and higher transgene expression than ssAAV2/8. This improved transduction efficiency might facilitate the development of improved gene therapy protocols for inherited retinal degenerations, particularly those caused by defects in photoreceptor-specific genes.
迄今为止,腺相关病毒(AAV)载体是唯一已被证明能有效转导光感受器细胞的基因治疗载体,因此已成为眼部转导最常用的载体。多种AAV血清型已在眼部进行了评估,其中第一种是AAV2,它能够转导光感受器、视网膜色素上皮(RPE)和视网膜神经节细胞。AAV血清型1和4,以及用这些衣壳假型化的AAV2,仅转导RPE。AAV血清型5和AAV2/5转导光感受器以及RPE,但不转导视网膜神经节细胞。在此,我们通过玻璃体内、视网膜下和前房内注射AAV2/8绿色荧光蛋白,评估了新型血清型AAV2/8转导成年小鼠视网膜各种眼部组织的能力。我们还确定了血清型2/2、2/5和2/8的自我互补AAV(scAAV)载体的动力学和效率,并将它们与单链AAV(ssAAV)进行了比较。我们发现,ssAAV2/8比ssAAV2/2和ssAAV2/5更有效地转导光感受器和RPE,并且scAAV2/8比ssAAV2/8起效更快且转基因表达更高。这种提高的转导效率可能有助于开发针对遗传性视网膜变性,特别是由光感受器特异性基因缺陷引起的遗传性视网膜变性的改进基因治疗方案。