一种可视化追踪卵泡细胞发育的卵巢早衰小鼠模型

Ovarian diseases such as premature ovarian failure are serious threats to women's health in modern society. In recent decade, the incidence of ovarian diseases is continuous increasing in China, however, there is still a lack of effective early diagnosis and treatment against those diseases. The main reason of this contradiction is that the pathogenesis and inner machinisms of ovarian diseases are still not clear, and the lack of effective animal model to study. Meanwhile, the developmental fate of granulosa cells, which are the major type of female endocrine cells, is closely related to the physiological and pathological changes in ovary. However, there is no proper animal model to trace the development of granulosa cells till now. The current project is based on the previous results and experimental experience on animal models in our group in last decade, and the purpose of the project is to create an inducible, granulosa cell dervied ovarian disease mouse model which is possible to perform the cell lineage tracing in study. By combining several genetically modified mouse strains, a granulosa cell derived premature ovarian failure could be induced by tamoxifen injection, and the developmental fate of both normal and abnormal granulosa cells could be traced by a multifluorescent reporter system. After the model established, a standard inducible protocol will be normalized in the project to make those ovarian disease mouse models standardized and reproducible. Finally, several exogenous factors such as stress and alcohol will be introduced to our model, and the etiological and pathological process of disease will be monitored in molecular and cellular levels to create an ideal model for ovarian disease investigation.

卵巢疾病如卵巢早衰严重威胁女性健康。近十年来，我国育龄女性卵巢疾病发病率不断攀升，却缺乏有效治疗方法。其主要原因在于对卵巢疾病的发病机制尚不完全清楚，并缺乏有效的动物模型进行实验研究。而在卵巢中，主要的内分泌细胞—颗粒细胞的发育命运无疑与卵巢生理病理变化密切相关。本项目中，申请人基于前期工作中所获得的基础研究结果及动物模型工作经验，将多种基因修饰动物加以结合，拟构建一种通过诱导敲除卵巢颗粒细胞基因，进而导致卵巢早衰的小鼠模型；同时，通过引入多重荧光报告小鼠，获得可在病理条件下对卵巢早衰中病变细胞直观进行追踪的卵巢体细胞疾病小鼠模型。在完成模型构建的基础上，进一步规范化诱导发病条件，并引入多种外源卵巢疾病诱导因素，对模型中的病程变化及病因在分子、细胞水平进行深入监控，最终期望为卵巢疾病的致病因素与治疗手段提供一个可深入研究和探索的，具有高度可塑性的模型动物研究平台。

卵巢疾病严重威胁女性健康，而近年来我国育龄女性卵巢相关疾病的发病率不断攀升，但缺乏有效的治疗方法，其中主要原因之一在于基础研究中对卵巢中相关细胞发生发育以及发病过程中的机制不完全清楚。同时，由于卵巢发育过程复杂，细胞种类繁多，因而相关探索对于在体研究非常依赖，但有效的动物模型却始终缺乏。在本项目中，我们将卵巢重要的细胞类型颗粒细胞条件性基因突变小鼠模型，与内源性荧光报告小鼠模型相互结合，构建了多种具有卵巢早衰病征，并可追踪卵巢颗粒细胞发育命运的小鼠模型。在模型构建完成的基础上，我们的工作详细的探索了卵巢早衰过程中突变颗粒细胞的发育分化命运，并发现了突变颗粒细胞在模型生命中期可转变为癌细胞，并对其中的相关机制进行了深入的解析。相关研究工作目前已被接收，即将发表。同时，在上述模型的基础上，我们利用多种条件性敲除动物模型，构建了基于基因修饰的诱导性卵巢早衰模型，并利用上述模型揭示了卵泡激活多态性的调控机制。总而言之，在项目支持下，本研究依照计划构建了多种先进的卵巢相关疾病研究动物模型，并对卵巢颗粒细胞相关疾病的发病机制进行了深入的揭示，项目目标全部完成。