BNC1调节卵母细胞发生发育与凋亡闭锁的机制研究

Premature ovarian failure (POF) is defined as loss of ovarian function in female before 40 years of age. Although the majority of POF are related to genetic mutation or genetic predisposition, the exact pathogenesis is unknown yet. In previous studies, we analyzed the collected clinical data and blood samples from hereditary or sporadic POF cases via genome-wide association study and exome sequencing, and became the first in the international arena to document BNC1 mutation as a possible genetic pathogenesis for POF. To further clarify BNC1 mutation as a causative mechanism for POF, we plan to analyze the reproductive endocrinology and ovarian aging phenotype, oocyte development or apoptosis, tissue expression profile chip, protein interaction and signal pathway via BNC1 target mutation animal, BNC1 Knockout animal and cellular targeted mutation model research，which help exploring the crucial step and key regulatory factors of BNC1 mutation inducing POF. What's more, methods such as protein crystallization and X-ray diffraction will be used to reveal BNC1 protein structure and to analyze the associated function. Our original research will sheds light on the understanding of the balance between oocytogenesis and oocyte atresia, and BNC1 mutation as a causative factor of POF at the molecular regulatory level, and also provide valuable theoretical basis for early intervention and genetic blockade of POF. Our study contributes to improving female reproductive health, lowering related health-care costs, and has both scientific value and translational medical significance.

卵巢早衰（POF）指女性40岁前发生卵巢功能衰竭，其发病与基因突变有关，但确切机制尚不明确。本课题组前期通过全基因组关联分析、外显子组捕获、二代测序方法分析遗传性POF家系及散发患者临床资料与血样，发现Basonuclin 1（BNC1）突变为人类POF新致病基因突变。本研究拟进一步采用BNC1靶向突变/敲除小鼠模型、体外细胞定点突变模型，通过生殖内分泌/卵巢衰老表型、卵子发生及闭锁观察、组织表达谱芯片检测、交互作用蛋白及其信号通路，研究BNC1对卵母细胞发生发育/凋亡闭锁的调节、及其影响卵巢衰老的关键步骤及调控因子；通过蛋白结晶、X衍射揭示BNC1蛋白结构，并进行功能关联分析。本原创性研究有助阐明BNC1调节卵子发生与闭锁平衡、及BNC1突变致POF的分子调控机制，为寻求相关早期干预和遗传阻断提供有价值的理论依据，有助促进女性生殖健康，降低相关医疗损耗，兼具重大科研价值和转化医学意义。

卵巢早衰定义为女性在40岁前发生卵巢功能衰竭，原发或继发闭经（至少4个月），并伴2次或以上血清卵泡刺激素上升和雌激素水平降低，严重影响患者生活质量及生殖健康。卵巢早衰的病因复杂，目前尚不明确，极有可能与遗传基因有关。本课题组前期在一个大型中国人遗传性卵巢早衰家系中成功筛选鉴定了一个新的致病基因Basonuclin1（BNC1）突变，并在后续82例散发性卵巢早衰患者中亦发现4例BNC1点突变。后续通过体外细胞靶向突变、基因敲减、在体靶向突变动物模型实验、突变蛋白结构预测等，进行突变蛋白结构与功能验证，确证BNC1突变导致蛋白入核障碍、卵子减数分裂异常、卵巢早衰表型发生，同时利用转录组学、染色质免疫共沉淀、生物信息学分析等探究BNC1下游目标基因以及相关通路，从而阐明BNC1靶向突变致卵巢早衰的分子机制。本原创性研究有助解析卵巢早衰的发病机理，为早期干预卵巢早衰和遗传阻断提供理论基础，促进生殖健康，提高女性生活质量，兼具重大科研价值和社会意义。