单细胞分辨率解析皮肤干细胞转分化生殖细胞的转录调控机制

利用小鼠多能性干细胞获得功能性配子已有报道，但是，多能性干细胞在临床应用上受到材料来源、致瘤性、伦理和免疫排斥等限制。相比较而言，皮肤来源干细胞（SDSCs）是一种非常有应用前景的干细胞类型。我们前期的研究发现SDSCs在向生殖细胞转分化过程中还存在效率低，减数分裂启动异常等问题。鉴于此，本项目拟利用scRNA seq技术，结合SDSCs向生殖细胞体外分化的实验平台，系统阐述SDSCs在“SDSCs-外胚层样细胞-原始生殖细胞样细胞-精原干细胞样细胞”转分化过程中的转录调控机制。借助与体内皮肤组织、外胚层细胞、原始生殖细胞以及精原干细胞单细胞图谱的比较分析，明确SDSCs来源的生殖细胞与体内生殖细胞之间基因表达、核心调控网络以及关键转录因子之间的差异，进而阐明SDSCs分化的生殖细胞无法继续分化形成功能性配子的原因。本项目将为今后利用SDSCs获得功能性配子的研究提供坚实的理论基础。

Obtaining functional gametes from pluripotent stem cells has been reported in mice. However, the current clinical application of pluripotent stem cells is limited due to material sources, tumorigenicity, ethics, and immune rejection. In comparison, skin-derived stem cells (SDSCs) are a kind of promising stem cell type. Our previous work has demonstrated that the differentiation of skin-derived stem cells into germ cells still faces low efficiency and abnormal meiosis initiation. Given these shortcomings, the current project intends to use single-cell transcriptome sequencing technology, combined with the experimental platform for the in vitro differentiation of skin-derived stem cells into germ cells, to analyze the detailed molecular mechanism underlying the transdifferentiation of SDSCs - Epiblast like cells – Primordial germ cell-like cells - Spermatogonia stem cell-like cells. Based on the comparison with in vivo skin tissues, epiblast cells, primordial germ cells, and spermatogonia stem cells, we hope to identify differences in gene expression, core regulatory networks, and key transcription factors between SDSCs-derived germ cells and in vivo germ cells, thus further demonstrating the reasons why SDSCs derived germ cells failed to differentiate into functional gametes. This project will provide a solid theoretical basis for future research on obtaining functional gametes from SDSCs.