Single Cell Transcriptome Analysis of Regenerating RGCs Reveals Potent Glaucoma Neural Repair Genes [First_batch_sur_RGCs]. Single Cell Transcriptome Analysis of Regenerating RGCs Reveals Potent Glaucoma Neural Repair Genes [First_batch_sur_RGCs]

Axon regeneration holds great promise for neural repair of CNS axonopathies, including glaucoma. Pten deletion in retinal ganglion cell (RGC) promotes potent optic nerve regeneration, but only a small population of Pten-null RGCs are actually regenerating RGCs (regRGCs); most surviving RGCs (surRGCs) remain non-regenerative. Here we developed a strategy to specifically label and purify regRGCs and surRGCs respectively from the same Pten deletion mice after optic nerve crush, in which they differ only in their regeneration capability. Smart-Seq2 single cell transcriptome analysis revealed novel regeneration-associated genes that significantly promote axon regeneration. The most potent of these, Anxa2, acts synergistically with its ligand tPA in Pten deletion-induced axon regeneration. Anxa2, its downstream effector ILK, and Mpp1 dramatically protect RGC somata and axons and preserves visual function in a clinically relevant model of glaucoma, demonstrating the exciting potential of this innovative strategy to identify novel effective neural repair candidates. Overall design: RGCs mRNA profiles of Pten knock out mice

轴突再生（Axon regeneration）在包括青光眼在内的中枢神经系统轴突病变的神经修复领域具有重大应用前景。视网膜神经节细胞（retinal ganglion cell, RGC）中的Pten敲除可强效促进视神经再生，但仅有极少量Pten缺失型RGC能够成为再生型RGC（regenerating RGCs, regRGCs）；多数存活的RGC（surviving RGCs, surRGCs）仍不具备再生能力。本研究开发了一种分选策略，可在视神经钳夹损伤后的同一批Pten敲除小鼠中，分别特异性标记并纯化仅再生能力存在差异的regRGCs与surRGCs。Smart-Seq2单细胞转录组分析揭示了一批可显著促进轴突再生的新型再生相关基因。其中作用最为强效的Anxa2，可与其配体tPA在Pten缺失诱导的轴突再生过程中发挥协同作用。Anxa2、其下游效应分子ILK以及Mpp1可在临床相关性青光眼模型中，有效保护RGC胞体与轴突并维持视觉功能，证实了该创新性策略用于筛选新型高效神经修复候选靶点的巨大潜力。总体实验设计：Pten敲除小鼠的RGC mRNA表达谱。