Molecular Discovery for Optic Nerve Regeneration

摘要 虽然成熟的视网膜神经节细胞（RGC）在正常情况下不能再生轴突， 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，分子和生理操纵，使一些RGC， 再生从眼睛到大脑的受损轴突。然而，尽管有这些努力， 在视神经损伤后存活并成功再生轴突进入大脑的RGC 仍然很小，从而限制了有意义的视力恢复。拟议的研究将联合收割机 一个强大的促再生疗法与新的转录组学和蛋白质组学方法， 尖端的生物信息学方法，以确定新的转录本和蛋白质与 启动和执行一个成功的再生程序。我们会调查 基因表达，蛋白质翻译和蛋白质转运的变化顺序 再生视神经作为成熟的RGC经历从正常完整状态到正常的神经节的转变。 稳健的生长状态，鉴定RGC亚群中选择性表达的转录物和蛋白质 它成功地将轴突延伸到神经中，并以RGC亚型为特征， 最有可能再生轴突。100-150个最重要的候选基因被鉴定出来， 发现阶段将测试它们在免疫纯化的RGC中促进轴突生长的能力 在文化，并从中间筛选主要候选人将测试他们的能力， 无论是在分离状态还是在体内， 与已建立的促再生疗法相结合。后面的研究将调查 将轴突定位到适当的中央视觉核团和视觉恢复测试。集成 这里提出的方法直接解决了识别新的分子靶点的目标， 在视觉系统受伤后建立视觉回路。

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.