支持细胞外泌体来源的miR-132-3p在精原干细胞自我更新中的作用及调控机制研究

miR-132-3p was significantly rich in SCOS’s exosomes by miRNA sequencing. Over expression of miR-132-3p inhibited expression of TAF4B. Moreover, bioinformatics analysis and reporter gene assays revealed that TAF4B was the target gene of miR-132-3p. The results suggested that SCOS sertoli cells may secrete exosomes containing miR-132-3p to inhibit self-renewal of SSCs by down regulating TAF4B expression. Our study is designed to verify the relationship between SCOS and miR-132-3p expression in exosomes secreted by sertoli cells. Besides, the mechanisms of miR-132-3p, transported by exosomes derived from sertoli cells, on regulating TAF4B in SCOS will be investigated by inhibiting the release of exosomes, overexpression and knocking-down miR-132-3p, and rescue experiments. To date, there has been no relevant report about sertoli cells regulating SSCs through exosomes，and also no relevant study about miR-132-3p regulating SSCs. Our study is expected to provide new targets for the prevention and treatment of SCOS.

miR-132-3p是我们通过miRNA测序技术筛选到的，显著高表达于SCOS病人支持细胞外泌体的miRNA；过表达miR-132-3p可抑制SSCs自我更新重要分子TAF4B的表达，生物信息学分析和报告基因实验均证实TAF4B为miR-132-3p的靶基因。因此，我们认为SCOS病人支持细胞可能通过外泌体转运miR-132-3p至SSCs，下调TAF4B的表达，调控SSCs的自我更新，参与SCOS的发生。本研究拟进一步阐明SCOS病人外泌体中miR-132-3p与SCOS患病的关系；通过抑制外泌体形成、过表达/沉默外泌体中miR-132-3p及挽救实验，系统评估外泌体中miR-132-3p调控TAF4B影响SSCs自我更新的分子机制。目前尚未见支持细胞通过外泌体影响SSCs功能的报道，也未见miR-132-3p参与调节SSCs功能的报道，本研究如获成功，有望为SCOS防治提供新靶标。

无精子症是引起男性不育的重要原因之一，唯支持细胞综合症是无精子症中最严重的一种类型，约占无精子症的13%-16%。唯支持细胞综合症生精障碍的病理生理机制尚未完全明确，本研究在前期工作的基础上进行了后续功能性研究，实时定量PCR技术证实了miR-132-3p在唯支持细胞综合症患者支持细胞外泌体中的表达水平显著高于对照组，通过体外抑制外泌体中miR-132-3p，发现低表达miR-132-3p能够显著上调精原干细胞的自我更新水平并降低其分化水平，体外细胞计数及EdU实验均证实低表达miR-132-3p能够显著上调精原干细胞增殖水平。此外，为了扩展对唯支持细胞综合症遗传学病因的认识，我们对唯支持细胞综合症患者进行了全基因组测序及生物信息学分析，初步揭示了唯支持细胞综合症新的遗传学病因。本研究具有源头创新性，有助于阐明唯支持细胞综合症生精障碍的机制，为其治疗提供新的线索和靶标。