Data_Sheet_3_A Subpopulation of Schwann Cell-Like Cells With Nerve Regeneration Signatures Is Identified Through Single-Cell RNA Sequencing.xlsx

Schwann cell-like cells (SCLCs) derived from human amniotic mesenchymal stem cells (hAMSCs) have been shown to promote peripheral nerve regeneration, but the underlying molecular mechanism was still poorly understood. In order to investigate the heterogeneity and potential molecular mechanism of SCLCs in the treatment of peripheral nerve regeneration at a single cell level, single-cell RNA sequencing was applied to profile single cell populations of hAMSCs and SCLCs. We profiled 6,008 and 5,140 single cells from hAMSCs and SCLCs, respectively. Based on bioinformatics analysis, pathways associated with proliferation, ECM organization, and tissue repair were enriched within both populations. Cell cycle analysis indicated that single cells within these two populations remained mostly in the G0/G1 phase. The transformation of single cells from hAMSCs to SCLCs was characterized by pseudotime analysis. Furthermore, we identified a subpopulation of SCLCs that highly expressed genes associated with Schwann cell proliferation, migration, and survival, such as JUN, JUND, and NRG1., Genes such as PTGS2, PITX1, VEGFA, and FGF2 that promote nerve regeneration were also highly expressed in single cells within this subpopulation, and terms associated with inflammatory and tissue repair were enriched in this subpopulation by pathway enrichment analysis. Our results indicate that a subpopulation of SCLCs with nerve regeneration signatures may be the key populations that promote nerve regeneration.

由人羊膜间充质干细胞（human amniotic mesenchymal stem cells, hAMSCs）诱导获得的施万细胞样细胞（Schwann cell-like cells, SCLCs）已被证实可促进外周神经再生，但其潜在分子机制仍未被充分阐明。为在单细胞层面探究施万细胞样细胞介导外周神经再生治疗的异质性与潜在分子机制，本研究采用单细胞RNA测序（single-cell RNA sequencing）技术，对人羊膜间充质干细胞与施万细胞样细胞的单细胞群体进行转录组谱分析。本研究分别获取了来自人羊膜间充质干细胞的6008个单细胞，以及来自施万细胞样细胞的5140个单细胞进行分析。基于生物信息学分析，两类细胞群体均富集到与细胞增殖、细胞外基质（extracellular matrix, ECM）重塑、组织修复相关的信号通路。细胞周期分析结果显示，两类群体中的单细胞大多停滞于G0/G1期。本研究通过拟时序分析（pseudotime analysis），刻画了人羊膜间充质干细胞向施万细胞样细胞转化的单细胞动态过程。此外，本研究鉴定出一类施万细胞样细胞亚群，该亚群高表达与施万细胞增殖、迁移及存活相关的基因，例如JUN、JUND与NRG1；该亚群的单细胞同时高表达PTGS2、PITX1、VEGFA与FGF2等可促进神经再生的基因，通路富集分析（pathway enrichment analysis）显示，该亚群显著富集到与炎症反应及组织修复相关的功能条目。本研究结果表明，携带神经再生特征的施万细胞样细胞亚群，或是介导外周神经再生的关键细胞群体。