BMP4信号通路在卵巢生殖干细胞分化过程中的作用及其机制研究

卵巢衰老是严重危害女性健康的重要因素。卵巢生殖干细胞（oogonial stem cells,OSCs）定向分化为卵母细胞可望成为延缓卵巢衰老的新策略。OSCs如何分化为卵母细胞的分子机制尚不清楚。我们前期已建立小鼠及人的OSCs细胞系，并发现BMP4因子可以促进OSCs分化；对添加BMP4因子前后的OSCs进行转录组测序，发现与卵子形成相关的基因（Sohlh1，Sohlh2，Lhx8）具有显著表达差异。我们推测BMP4可能通过调控上述基因促进鼠与人OSCs的分化。本研究拟采用DDX4启动子调控的绿色荧光蛋白标记的卵巢生殖干细胞(DDX4/EGFP-OSCs)，联合生物信息学分析预测、染色质免疫沉淀技术（ChIP）、CRISPR/Cas9基因编辑技术等探讨BMP4下游关键基因及其调控分子，从而构建BMP4信号通路调控卵巢生殖干细胞分化的网络图，为临床运用OSCs延缓卵巢衰老提供理论依据。

Ovarian aging is an important factor that seriously impairs women's health. The differentiation of oogonial stem cells(OSCs) into oocytes is expected to be a new strategy to delay ovarian aging. The molecular mechanism of how OSCs differentiate into oocyte remains unclear. We have established the mouse and human OSCs cell lines, and found that the BMP4 factor can promote differentiation of OSCs. And we have done transcriptome high-throughput sequencing on the OSCs before and after the treatment of BMP4, finding that the genes Sohlh1, Sohlh2, Lhx8, which are related to the formation of eggs have a significant difference. We hypothesized that BMP4 might contribute to the differentiation of OSCs by regulating the genes above. In this study, using the OSCs modern in which GFP gene was controlled under the promoter of DDX4 gene, and combined with biological information analysis prediction, chromatin immunoprecipitation assay (CHIP) and CRISPR / cas9 gene editing technology. We plan to explore the downstream key genes and molecular regulation of the BMP4 signaling pathway, so as to construct the network diagram about that how the BMP4 signaling pathway regulates the differentiation of OSCs, thus providing theoretical basis for the clinical application of OSCs for delaying ovarian aging.