小分子W7Y24促进人类多能干细胞分化为生殖细胞的分子机制研究

建立一个高效的体外分化体系以获得原始生殖细胞及性原细胞是实现体外配子发生的重要步骤，但目前人类生殖细胞的体外分化仍面临效率低、不同步及早期分化滞留的问题。在前期研究中，我们建立了一个可持续量化检测生殖细胞发生的报告基因系统，并进行了高通量小分子化合物筛选，发现JAK3强效抑制剂小分子W7Y24能显著提高干细胞分化为生殖细胞的效率，但其作用机制仍不清楚。为了进一步揭示W7Y24促进生殖细胞分化的分子机理，在本项目中我们将通过鉴定W7Y24诱导分化的细胞属性，锁定W7Y24的靶向信号通路及效应转录因子，并结合模式生物分子机制的保守性的验证，从而揭示其未知的人类生殖细胞分化的分子机制。相关结果将极大地推进人类体外配子发生研究，扩展我们对人类生殖细胞发育和疾病发生的分子机制的认知和理解，为不育不孕等疾病的治疗提供新的靶点及工具。

In vitro gametogenesis from human pluripotent stem cells (hPSCs) could serve as a genetic model to characterize the fundamental basis for human germ cell development, as well as an alternative source of gametes for the treatment of human infertility. In spite of many successes in association with the development of stem cell biology, methods of human germ cell differentiation in vitro remain inefficient, asynchronous and unable to proceed beyond the early hPGCs stage. The development of a protocol or strategy to efficiently differentiate hPSCs towards later stage hPGCs that are comparable to gonocytes is essential for the achievement of in vitro gametogenesis. Here, we developed a human germ cell-specific quantification reporter assay that we can monitor the derivation of human germ cells non-invasively and quantitatively. We performed a high throughput screening and identified a small molecule that can enhance the differentiation from hPSCs to hPGCs significantly. For this project, we are going to comprehensively examine the target pathways and transcription factors of the small molecule. We anticipate that with the successful completion of this project, we will have a robust genetic system towards in-vitro gametogenesis as well as a greater understanding of the human-specific mechanism of germ cell development and infertility. Ultimately, it will provide novel therapeutic approaches for treatment of infertility.