The role of Notch in establishing ovarian interstitial cells

项目摘要 卵巢功能障碍是美国女性不孕不育的主要原因之一 各州。卵巢疾病和功能障碍，如多囊卵巢综合征，产生雄激素的肿瘤， 原发性卵巢功能不全或高雄激素症会导致女性不孕。通常这些疾病会导致 来自卵巢形成的缺陷，特别是在胎儿时期。胎儿卵巢的形成需要 三种主要细胞谱系：生殖细胞谱系形成卵母细胞，支持细胞谱系形成颗粒细胞 细胞和间质细胞谱系有助于类固醇生成和非类固醇生成基质细胞群。 卵巢中的生殖细胞和支持细胞谱系被广泛研究，因为这两种细胞 类型形成卵泡，卵泡是卵巢的功能单位。相反，间质细胞谱系的研究较少。 尽管如此，它的功能障碍与卵巢疾病有关。关于卵巢间质的一个关键知识缺口 细胞是其具体化和形成所必需的细胞动力学。此应用程序的主要目标是 填补这一知识空白，为疾病诊断、治疗和预防方面的进步奠定基础 使用鼠标作为模型。 在申请者在姚实验室做博士后的第一年里，他们发现了一种独特的 卵巢中激活Notch并在成年期特异发育成间质细胞的细胞群 提示Notch可能在卵巢细胞发育中发挥作用。这些发现导致了中央 这一建议的假设：缺口激活建立了胎儿卵巢的间质细胞谱系。 目的1将研究卵巢间质细胞的发育是否需要Notch激活。这将是 通过使用条件和可诱导的遗传小鼠模型来沉默Notch在小鼠体内的激活 祖细胞间质细胞群。目标2将测试异位Notch激活是否会改变支持 通过使用分离的、有条件的和可诱导的小鼠遗传模型将细胞转化为间质细胞 专门针对支持细胞群体。所有遗传小鼠模型的卵巢都将被解剖， 进行形态分析，并进行单细胞RNA测序分析。AIM 3将调查如何 支持细胞和间质细胞群通过Notch途径相互作用，以确定它们的独特身份。这 将通过对血统追踪的卵巢进行单细胞测序来完成。这项提案将解决 Notch激活参与了间质细胞命运的决定。从这项建议中获得的知识 将有助于更好地理解胚胎细胞是如何获得其祖细胞身份的 推动卵巢间质细胞发育。有了这些信息，未来的研究可以直接解决影响 环境或其他压力因素对这些过程的影响。最后，这些结果可能会被翻译成人类 涉及卵巢疾病和功能障碍的健康风险。

Project Summary Ovarian dysfunction is one of the leading causes behind women experiencing infertility in the United States. Ovarian diseases and dysfunctions such as polycystic ovarian syndrome, androgen-producing tumors, primary ovarian insufficiency, or hyperandrogenism contribute to infertility in women. Often these diseases result from defects in ovary formation, particularly during fetal life. Formation of the fetal ovary requires the presence of three main cell lineages: the germ cell lineage forms oocytes, the supporting cell lineage becomes granulosa cells, and the interstitial cell lineage contribute to steroidogenic and non-steroidogenic stromal cell populations. Germ cell and supporting cell lineages in the ovary are studied extensively due to the fact that these two cell types form the follicles, the functional unit in the ovary. On the contrary, the interstitial cell lineage is less studied despite that its dysfunction is associated with ovarian diseases. A critical knowledge gap on ovarian interstitial cells is the cellular dynamics necessary for its specification and formation. The main goal of this application is to fill this knowledge gap and lay the foundation for advances in disease diagnosis, treatment, and prevention by using the mouse as the model. Within the applicant’s first year as a post-doctoral fellow in the Yao Laboratory, they discovered a unique population of cells in the ovary that activate Notch and specifically develop into interstitial cells in adulthood indicating that Notch may play a role in ovarian cell development. These findings lead to the central hypothesis of this proposal: Notch activation establishes the interstitial cell lineage in the fetal ovary. AIM 1 will investigate whether ovarian interstitial cell development requires Notch activation. This will be accomplished by using a conditional and inducible genetic mouse model that silences Notch activation in the progenitor interstitial cell population. AIM 2 will test whether ectopic Notch activation will transform the supporting cells into interstitial cells through the use of separate, conditional, and inducible genetic mouse model that specifically targets the supporting cell population. Ovaries from all of the genetic mouse models will be dissected, morphologically analyzed, and subjected to single cell RNA sequencing for analysis. AIM 3 will investigate how supporting cell and interstitial cell populations interact via the Notch pathway to define their unique identity. This will be accomplished by performing single cell sequencing on lineage traced ovaries. This proposal will address the involvement of the Notch activation in interstitial cell fate determination. Knowledge gained from this proposal will provide a better understanding of how embryonic cells gain their progenitor identity and the mechanisms driving ovarian interstitial cell development. With this information, future studies can directly address the impacts of the environment or other stressors on these processes. Finally, these results may be translated to human health risks involving ovarian disease and dysfunction.