GDF-Sox9/PTEN信号轴调控（类）角膜内皮祖细胞活化增殖与间质化平衡的作用与机制研究

目前针对角膜内皮失代偿所致角膜盲唯一治疗方法是角膜移植。供体和移植后角膜内皮细胞活性制约其发展。（类）角膜内皮祖细胞的增殖与间质化（EnMT）平衡是成功获得移植细胞和移植后体内存活的关键。生长分化因子（GDF）通过拮抗TGF-β双向调控祖细胞增殖，我们前期研究发现，小鼠角膜内皮修复时GDF、Sox9表达升高，胚胎干细胞诱导分化为角膜内皮细胞过程中GDF、PTEN表达与增殖、EnMT相关。我们提出假说：在（类）角膜内皮祖细胞体外诱导或体内活化时，GDF时序性激活Sox9和PTEN表达，Sox9启动祖细胞增殖分化，至成熟时，PTEN抑制细胞过度增生与EnMT。计划采用全转录组学、谱系或活体示踪、Myc-GFP-TET-ON 技术等检测和干预手段阐明GDF-Sox9/PTEN信号轴在（类）角膜内皮祖细胞增殖与间质化平衡的作用与机制，为促进在体角膜内皮细胞自发修复、提高种子细胞质量提供新方案。

At present, the only clinical treatment for corneal blindness caused by corneal endothelial decompensation is corneal transplantation. However, it’s the main limitation that the activity of donor and post-transplant corneal endothelial cells. Furthermore, the balance of proliferation and endothelial mesenchymal transformation in (Like) corneal endothelial progenitor cells is the key to obtain and maintain survival successfully of the corneal endothelial cells in vivo. GDF regulates the cells proliferates by antagonalizing TGF-β.We found that the expressions of GDP and Sox9 in corneal endothelial proliferation were increased in the repairing of mice, and GDF and PTEN were closely related to the proliferation of CECs and EnMT during ESCs induced differentiation. Therefore, we proposed that GDF expression is increased, when corneal endothelial progenitor cells are induced in vitro or activated in vivo.Then Sox9 and PTEN transcription are sequentially activated, and the proliferation of endothelial progenitor cells are initiated by Sox9. When the cells are differentiated and mature, PTEN expression is increased to inhibit the overproliferation of HCEnCs and EnMT.In this subject, we planned to clarify the role of GDF-Sox9 /PTEN axis in the repairment of corneal endothelial cell damage, using the techniques including whole transcriptomics analysis, lineage or live tracking, Myc-GFP-TET-ON technology, retrovirus overexpression and SiRNA knockdown. In the end we wish that we can propose and validate a plan to induce the self-repairment of corneal endothelial cell injury, improve the quality of corneal endothelial seed cells for transplantation and inhibit the onset of EnMT.