Song Hongman, Bush Ronald A, Zeng Yong, Qian Haohua, Wu Zhijian, Sieving Paul A
Section for Translational Research on Retinal and Macular Degeneration, National Institute on Deafness and Other Communication Disorders, Bethesda, MD 20892, USA.
National Eye Institute, NIH, Bethesda, MD 20892, USA.
Mol Ther Methods Clin Dev. 2018 Dec 21;13:77-85. doi: 10.1016/j.omtm.2018.12.006. eCollection 2019 Jun 14.
Adeno-associated virus (AAV) vector-mediated gene delivery is a promising approach for therapy, but implementation in the eye currently is hampered by the need for delivering the vector underneath the retina, using surgical application into the subretinal space. This limits the extent of the retina that is treated and may cause surgical injury. Vector delivery into the vitreous cavity would be preferable because it is surgically less invasive and would reach more of the retina. Unfortunately, most conventional, non-modified AAV vector serotypes penetrate the retina poorly from the vitreous; this limits efficient transduction and expression by target cells (retinal pigment epithelium and photoreceptors). We developed a method of applying a small and safe electric current across the intact eye for a brief period following intravitreal vector administration. This significantly improved AAV-mediated transduction of retinal cells in wild-type mice following intravitreal delivery, with gene expression in retinal pigment epithelium and photoreceptor cells. The low-level current had no adverse effects on retinal structure and function. This method should be generally applicable for other AAV serotypes and may have broad application in both basic research and clinical studies.
腺相关病毒（AAV）载体介导的基因递送是一种很有前景的治疗方法，但目前在眼部应用时，由于需要通过手术将载体注入视网膜下间隙，在视网膜下方递送载体，这限制了可治疗的视网膜范围，并且可能导致手术损伤。将载体递送至玻璃体腔会更可取，因为它的手术侵入性较小，并且能覆盖更多的视网膜区域。不幸的是，大多数传统的、未修饰的AAV载体血清型从玻璃体穿透视网膜的能力很差；这限制了靶细胞（视网膜色素上皮细胞和光感受器）的有效转导和表达。我们开发了一种方法，在玻璃体腔内注射载体后，对完整的眼睛短暂施加微弱且安全的电流。这显著提高了野生型小鼠玻璃体腔内注射后AAV介导的视网膜细胞转导效率，使视网膜色素上皮细胞和光感受器细胞中出现基因表达。这种低强度电流对视网膜结构和功能没有不良影响。该方法应普遍适用于其他AAV血清型，并且可能在基础研究和临床研究中都有广泛应用。
