卵母细胞成熟障碍是一种可带来沉重医疗负担的疑难生殖障碍性疾病，其精确发病机制至今不明。SETD2介导的H3K36me3修饰在DNA损伤修复中发挥重要功能，本研究前期发现小鼠卵母细胞体外成熟培养过程中MPF活化与SETD2表达水平之间存在相关性，Setd2敲除小鼠卵母细胞出现GV期阻滞及DNA损伤表型，卵母细胞SMART-Seq结果提示Setd2基因缺失可致DNA损伤修复通路下调。本研究拟进一步采用Setd2敲除小鼠模型、Setd2过表达/敲减细胞模型，观察卵子发育及DNA损伤修复表型，明确Setd2调控卵母细胞成熟的效应及机制，通过染色质免疫共沉淀、Western-blot、信号通路阻断等探索MPF/MAPK调控Setd2表达的关键步骤及调控因子，从分子、细胞、临床样本以及动物水平多层次明确SETD2介导MPF/MAPK调控卵母细胞成熟的分子网络，为卵母细胞成熟障碍提供潜在的干预靶点。

Oocyte maturation disorder is a complicated reproductive disorder, which added heavy burden to patients, families and even the society. SETD2 mediated H3K36me3 modification have been proved to play important roles in DNA damage response and repair. We found that MPF activation levels were relevant to the expression levels of SETD2 in mouse GV oocyte undergoing in vitro maturation culture in our previous study. Furthermore, SETD2 knockout mice oocytes present to be blocked by GV stage and have disorder in DNA damage repair. SMART- Seq also showed that SETD2 knock-out down-regulated DNA damage repair pathways and progesterone mediated oocyte maturation pathway in those oocytes. Based on the previous results, we plan to further clarify SETD2-mediated effects and molecular mechanisms of MPF/MAPK in the regulation of oocyte maturation, by constructing mouse/cell model with Setd2 knock-in and cell model with Setd2 knock-down. The phenotype including oocyte maturation and DNA damage repair of the animal model and cell models will be analyzed to confirm whether Setd2 defect will impair oocyte maturation. A series of experiments by real-time PCR, Western-blot , signal transduction pathway blockage and chromatin immunoprecipitation will be employed to explore the effects and mechanisms of MPF/MAPK in up-regulating Setd2, and to figure out the crucial steps, the key regulatory factors and the precise regulatory network of MPF/MAPK and Setd2 in oocyte maturation regulation both in vivo and in vitro at multiple levels from molecules, cells, clinical samples to animal models, thus providing potential intervention targets for oocyte maturation disorders.