The regulation of ovarian aging by H19 and let-7

项目摘要/摘要 第一胎母亲的平均年龄正在上升，这对产妇健康和女性健康造成了严重后果。 他们的后代。高龄产妇与出生缺陷、流产和不孕症密切相关。这些 不良的生殖结局往往可以追溯到卵泡和卵母细胞的数量和质量的缺陷。 在卵巢老化的情况下发生。卵泡数量的持续下降(即“卵巢储备”) 发生在生殖老化期间，因为原始卵泡池不断耗尽。此外，卵母细胞 随着时间的推移，卵泡和卵母细胞积累DNA损伤，质量会下降，这一过程甚至会加速 当卵巢暴露在性腺毒素中时会更快。因此，迫切需要更好地理解 控制卵泡和卵母细胞数量和质量以支持妇女健康的机制 和他们的孩子。非编码RNAs H19和let-7在哺乳动物发育中起重要作用，但作用很小 已知它们在卵泡生长和卵母细胞功能中的作用。我们发现了一个看似合理的 通过H19/let-7对基于非编码RNA调节卵泡健康的机制。我们之前展示了 H19结合并拮抗miRNA let-7。我们还证明了在没有H19的情况下，卵巢 AMH表达减少，卵泡募集加速，生育能力受损。我们有 观察到AMH有一个功能性的let-7结合位点，提示AMH的机制是由ncRNA介导的 H19通过LET-7进行监管。此外，我们的初步数据表明，人类对DNA损伤的反应发生了变化 没有H19病毒。因此，对H19和LET的作用有看似合理的机械洞察力和强有力的支持- 7在卵泡/卵母细胞募集和功能的调节中起重要作用。在目标1中，我们将确定H19在E2中的作用- AMH对卵泡数量的调节作用。在目标2中，我们将确定H19KO小鼠的卵巢 比它们的WT同龄人更容易受到DNA损伤。最后，对于目标3，我们将确定是否 H19KO小鼠卵泡发育和DNA损伤基因表达异常 通过let-7介导，并鉴定体细胞和卵母细胞转录组中与丢失相关的变化 H19.我们的方法是创新的，因为它代表着与现状的实质性偏离， 将非编码RNA(NcRNAs)定义为卵子数量和质量的主要调节因子，并有可能 为多种生殖疾病状态带来基于ncRNA的新型治疗方法。

PROJECT SUMMARY/ABSTRACT The mean age of first-time mothers is on the rise, with serious consequences for maternal health and that of their offspring. Older maternal age is strongly associated with birth defects, miscarriage, and infertility. These poor reproductive outcomes can frequently be traced back to defects in follicular and oocyte quantity and quality that occur with ovarian aging. A continuous decline in the quantity of ovarian follicles (i.e. the “ovarian reserve”) occurs during reproductive aging, as the pool of primordial follicles is continuously depleted. Additionally, oocyte quality declines as follicles and oocytes accumulate DNA damage over time, a process which accelerates even more rapidly when the ovary is exposed to gonadotoxins. There is therefore an urgent need to better understand the mechanisms that control follicular and oocyte quantity and quality in order to support the health of women and their children. The noncoding RNAs H19 and let-7 play essential roles in mammalian development, but little is known about their role in ovarian follicle growth and oocyte function. We have uncovered a plausible mechanism for noncoding-RNA-based regulation of follicular health via the H19/let-7 pair. We previously showed that H19 binds and antagonizes the miRNA let-7. We also demonstrated that in the absence of H19, ovarian AMH expression is decreased, follicular recruitment is accelerated, and fertility is compromised. We have observed that AMH has a functional let-7 binding site, suggesting a ncRNA-mediated mechanism for AMH regulation by H19 via let-7. Moreover, our preliminary data suggests altered response to DNA damage in the absence of H19. Thus, there is plausible mechanistic insight into, and strong support for, the role of H19 and let- 7 in the regulation of follicular/oocyte recruitment and function. In Aim 1, we will determine the role of H19 in E2- and AMH-mediated regulation of follicle quantity. In Aim 2, we will determine whether ovaries of H19KO mice are more susceptible to DNA damage than their WT counterparts. Lastly, for Aim 3, we will determine whether the abnormal follicular development and expression of DNA damage genes observed in H19KO mice is mediated via let-7 and identify changes in the transcriptome of somatic cells and oocytes related to loss of H19. Our approach is innovative because it represents a substantive departure from the status quo by defining noncoding RNAs (ncRNAs) as major regulators of oocyte quantity and quality, and has the potential to lead to novel, ncRNA-based treatments for a broad range of reproductive disease states.