Molecular Discovery for Optic Nerve Regeneration

摘要 尽管成熟的视网膜神经节细胞(RGC)通常不能再生轴突 视神经损伤，来自几个实验室的研究，包括参与这项合作的实验室， 已经确定了细胞、分子和生理操作，使一些RGC能够 将受损的轴突从眼睛再生到大脑。然而，尽管做出了这些努力，但 视神经损伤后存活并成功再生轴突进入大脑的视网膜节细胞 仍然很小，从而限制了有意义的视觉恢复。拟议的研究将结合 采用新的转录和蛋白质组学方法的强大的促再生疗法 尖端生物信息学方法识别与该基因相关的新转录本和蛋白质 启动并执行成功的再生程序。我们将调查时间 基因表达、蛋白质翻译和蛋白质向下运输的序列变化 再生的视神经作为成熟的视网膜节细胞经历了从正常完整状态到 强健的生长状态，鉴定RGC亚群中选择性表达的转录本和蛋白质 这成功地将轴突延伸到神经中，并用 再生轴突的潜力最大。100-150个最重要的候选基因在 发现阶段将测试它们促进免疫纯化的视网膜节细胞轴突生长的能力。 在文化方面，来自中间筛选的主要候选人将接受测试，以测试他们的能力 显著提高体内视神经再生水平，无论是孤立的还是 与已有的促再生疗法相结合。后一项研究将调查 将轴突定位于适当的中央视觉核团和视觉恢复测试。集成的 这里提出的方法直接解决了识别新的分子靶点以重新定位的目标。 在视觉系统受损后建立视觉回路。

Abstract Although mature retinal ganglion cells (RGCs) are normally unable to regenerate axons following optic nerve damage, studies from several labs, including those participating in this collaboration, have identified cellular, molecular, and physiological manipulations that enable some RGCs to regenerate injured axons from the eye to the brain. In spite of these efforts, however, the number of RGCs that survive after optic nerve injury and successfully regenerate axons into the brain remains small, thereby limiting meaningful visual recovery. The proposed research will combine a strong pro-regenerative therapy with novel transcriptomic and proteomic approaches and cutting-edge bioinformatic methods to identify new transcripts and proteins associated with the initiation and execution of a successful regenerative program. We will investigate the temporal sequence of changes in gene expression, protein translation, and protein transport down the regenerating optic nerve as mature RGCs undergo a transition from a normal intact state into a robust growth state, identify transcripts and proteins selectively expressed in the subset of RGCs that successfully extends axons into the nerve, and characterize the RGC subtypes with the highest potential to regenerate axons. 100-150 of the top candidate genes identified in the discovery phase will be tested for their ability to promote axon outgrowth in immunopurified RGCs in culture, and lead candidates from the intermediate screen will be tested for their ability to substantially augment levels of optic nerve regeneration in vivo, either in isolation or in combination with established pro-regenerative therapies. These latter studies will investigate the targeting of axons to appropriate central visual nuclei and tests of visual recovery. The integrated approach proposed here directly addresses the goal of identifying novel molecular targets to re- establish visual circuitry after injury to the visual system.