Department of Biological Sciences, Vanderbilt University, Nashville, TN, 37235, USA.
Department of Ophthalmology, Vanderbilt University Medical Center, Nashville, TN, 37235, USA.
Exp Eye Res. 2020 Nov;200:108254. doi: 10.1016/j.exer.2020.108254. Epub 2020 Sep 19.
Ongoing research using cell transplantation and viral-mediated gene therapy has been making progress to restore vision by retinal repair, but targeted delivery and complete cellular integration remain challenging. An alternative approach is to induce endogenous Müller glia (MG) to regenerate lost neurons and photoreceptors, as occurs spontaneously in teleost fish and amphibians. Extracellular vesicles (EVs) can transfer protein and RNA cargo between cells serving as a novel means of cell-cell communication. We conducted an in vivo screen in zebrafish to identify sources of EVs that could induce MG to dedifferentiate and generate proliferating progenitor cells after intravitreal injection into otherwise undamaged zebrafish eyes. Small EVs (sEVs) from C6 glioma cells were the most consistent at inducing MG-derived proliferating cells. Ascl1a expression increased after intravitreal injection of C6 sEVs and knockdown of ascl1a inhibited the induction of proliferation. Proteomic and RNAseq analyses of EV cargo content were performed to begin to identify key factors that might target EVs to MG and initiate retina regeneration.
正在进行的使用细胞移植和病毒介导的基因治疗的研究，通过视网膜修复来恢复视力方面取得了进展，但靶向输送和完全细胞整合仍然具有挑战性。另一种方法是诱导内源性 Muller 胶质细胞（MG）再生丢失的神经元和光感受器，就像在硬骨鱼和两栖动物中自发发生的那样。细胞外囊泡（EVs）可以在细胞之间转移蛋白质和 RNA 货物，作为一种新的细胞间通讯方式。我们在斑马鱼中进行了体内筛选，以鉴定可以诱导 MG 去分化并在向未受损的斑马鱼眼睛内注射后产生增殖祖细胞的 EV 来源。来自 C6 神经胶质瘤细胞的小细胞外囊泡（sEVs）在诱导 MG 衍生的增殖细胞方面最一致。C6 sEV 注射后 Ascl1a 的表达增加，而 Ascl1a 的敲低抑制了增殖的诱导。对 EV 货物含量的蛋白质组学和 RNAseq 分析进行了分析，以开始鉴定可能将 EV 靶向 MG 并启动视网膜再生的关键因素。
