卵巢特异性lncRNA OSTR对卵巢生殖干细胞的功能影响与相关应用研究

Clarifying the mechanisms of ovarian aging and exploring early biomarkers are of great significance. Others’ and our studies have shown that ovarian germline stem cell (OGSC) is a deciding factor for ovarian aging, while Hippo pathway plays an important role in the regulation of OGSGs function. Recently, we found that ovarian specific lncRNA OSTR can target the downstream transcription factor TEAD mRNA of the Hippo pathway, and its expression increased significantly in the animal model of premature ovarian insufficiency (POI). Thus, we hypothesize that OSTR may influence OGSCs’function and regulate ovarian aging through Hippo pathway. In this project, we will investigate the spatio- temporal expression of OSTR in OGSCs and its influence on the fate of OGSCs; reveal the role of OGSCs transplantation / ovarian micro-injection on the rescuing / recovery of physiological / pathological aging ovarian function through genetically manipulating OSTR; analyze the consistency of OSTR distribution in ovarian tissues and peripheral blood and explore the correlation between this consistency and the change of the common clinical biomarkers of ovarian aging, and further evaluate for its specificity and sensitivity, hoping the success of this project will provide valuable scientific basis for the diagosis and treatment of the related female reproductive disorders.

阐明卵巢衰老机理及探寻早期标志物具有重大现实意义。他人与我们的研究表明，卵巢生殖干细胞（OGSCs）是影响卵巢衰老的重要因素，而Hippo信号通路则是调控OGSCs功能的关键途径。新近我们又发现：卵巢特异性lncRNA OSTR靶向Hippo通路下游转录因子TEADmRNA，且在早衰卵巢中表达显著增加。故我们假设OSTR通过Hippo通路影响OGSCs功能并调控卵巢衰老。本课题拟以GFP转基因小鼠和人为实验对象，观察OSTR在OGSCs的时空表达谱变化及其对OGSCs命运的影响；揭示OSTR活性操控后的OGSCs移植/在体卵巢微注射对拯救生理/病理性衰老卵巢功能的作用；探讨OSTR在卵巢和外周血中分布的一致性及与常用卵巢衰老检测指标变化的时序相关性；临床资料进一步评估其作为卵巢衰老标记物的特异性与敏感性。本申请旨在为重塑和保护卵巢功能提供新靶点，为寻找卵巢衰老早期标记物提供实验与临床前证据

阐明卵巢衰老机理及探寻早期标志物具有重大现实意义。他人与我们的研究表明，卵巢生殖干细胞（OGSCs）是影响卵巢衰老的重要因素，而Hippo信号通路则是调控OGSCs功能的关键途径。然而，转录或转录后调控分子lncRNA是否可通过Hippo通路影响OGSCs功能并调控卵巢衰老并不清楚。本课题研究主要包含以下内容：①筛选、验证并鉴定衰老卵巢中与Hippo信号通路密切相关的lncRNAs，并观察OSTR在小鼠OGSCs的时空表达谱变化；②观察调控OSTR对OGSCs命运的影响，分析与Hippo通路相关关系；③观察在体卵巢微注射相关慢病毒对小鼠卵巢功能的影响和作用机制。研究结果发现：①在小鼠卵巢原始卵泡启动前后以及生理性衰老过程中，存在大量lncRNAs的差异性表达；②靶向Hippo-Tead2的OSTR（ENSMUST00000145581，简称lnc81）随着鼠龄增加表达量增加，且在OGSCs中的表达高于GCs中的表达并主要定位于细胞核；③lnc81通过Hippo-TEAD2-CCN1/2信号通路调控FGSCs和GCs的增殖凋亡；④在体卵巢微注射lnc81过表达载体可能诱发卵巢早衰。本研究首次筛选与鉴定出卵巢衰老密切相关的长链非编码RNA-lnc81，并证实lnc81不仅直接调控FGSCs的增殖、凋亡，还参与GCs调控FGSCs增殖、凋亡的过程，从而影响卵巢衰老。研究成果为深入探讨维持雌性卵巢生理功能的内在机制增添新资料，为重塑和保护卵巢功能提供新靶点。