人类早期胚胎体外发育阻滞过程中DNA主动去甲基化的调控规律及功能研究

DNA主动去甲基化是哺乳动物早期胚胎发育过程中极为重要的表观遗传重编程，该过程异常会导致胚胎体外培养发育过程的停滞，但是其在人类早期胚胎体外发育阻滞过程中的具体变化规律和调控特征还尚未完全清楚。因此，本项目以DNA主动去甲基化的重要中间产物5-醛基胞嘧啶（5-formylcytosine，5fC）为切入点，利用我们已经报道掌握的CELVER-seq技术，结合运用该技术绘制完成的人类早期胚胎体外正常发育过程中DNA主动去甲基化的特征图谱，进一步深入比较并揭示人类早期体外发育阻滞胚胎在DNA主动去甲基化方面的异常调控模式和特点。最后，对上述找出的在阻滞胚胎中的异常特征进行分子水平的验证，以及初步的功能性探索实验，阐明DNA主动去甲基化异常导致胚胎发育阻滞的机制。本项目将会加深对人类早期胚胎发育过程中分子调控规律的认识，为进一步改善胚胎体外培养体系，提高“试管婴儿”的成功率提供重要的理论支撑。

DNA methylation reprogramming plays a crucial role in epigenetic regulation during human early embryonic development, the dysfunction of this process might lead to embryonic arrest. However, the active DNA demethylation process during this crucial period remains largely unexplored. In this study, we use single-cell CLEVER-seq to quantify the 5-formylcytosine (5fC) level of human arrested preimplantation embryos and compare to figure out the abnormal DNA demethylation pattern during this period. Furthermore, we will check and identify candidate genes or elements with abnormal DNA demethylation pattern which might be associated with the development arrest. And then figure out the mechanism of abnormal DNA demethylation in development arrest. Collectively, our work offers new insights into the molecular basis of the complex demethylation process during human embryonic development and provides clues to improve the efficiency for in-vitro fertilization-embryo transfer (IVF-ET).