Deep scRNA sequencing reveals a broadly applicable Regeneration Classifier and implicates antioxidant response in corticospinal axon regeneration by Kim et al.2023 Samples

Despite substantial progress in understanding the biology of axon regeneration in the CNS, our ability to promote regeneration of the clinically important corticospinal tract (CST) after spinal cord injury remains limited. To understand regenerative heterogeneity, we conducted patch-based single cell RNA sequencing on rare regenerating CST neurons at high depth following PTEN and SOCS3 deletion. Supervised classification with Garnett gave rise to a Regeneration Classifier, which can be broadly applied to predict the regenerative potential of diverse neuronal types across developmental stages or after injury. Network analyses highlighted the importance of antioxidant response and mitochondrial biogenesis. Conditional gene deletion validated a role for NFE2L2 (or NRF2), a master regulator of antioxidant response, in CST regeneration. Our data demonstrate a universal transcriptomic signature underlying the regenerative potential of vastly different neuronal populations, and illustrate that deep sequencing of only hundreds of phenotypically identified neurons has the power to advance regenerative biology.尽管学界在解析中枢神经系统（Central Nervous System, CNS）轴突再生的生物学机制方面已取得显著进展，但在促进脊髓损伤后具有临床重要性的皮质脊髓束（corticospinal tract, CST）再生方面，我们的能力仍存在明显局限。为解析再生异质性，我们在PTEN与SOCS3基因敲除后，对少量罕见的再生型皮质脊髓束神经元开展了高深度的贴片式单细胞RNA测序。使用Garnett工具进行监督分类后，我们构建了再生分类器，该分类器可广泛用于预测不同发育阶段或损伤后各类神经元的再生潜能。网络分析结果凸显了抗氧化应答与线粒体生物发生的关键作用。条件性基因敲除实验验证了核因子红细胞2相关因子2（NFE2L2，又名NRF2）——一种抗氧化应答的核心调控因子——在皮质脊髓束再生中的功能。本研究数据表明，截然不同的神经元群体的再生潜能背后存在一套通用的转录组特征；同时证实，仅对数百个经表型鉴定的神经元开展深度测序，即可有力推动再生生物学领域的发展。