A Mouse Model to Demonstrate the Impact of Myometrial FSHR on Fertility

项目总结 卵泡刺激素受体(FSHR)在女性生育中的作用被认为是有限的 已知的卵巢FSHR的作用。我的实验室最近发表的研究对这一教条提出了挑战 通过展示卵巢外FSHR在女性生殖道中的表达 妇女和孕妇的胎盘和蜕膜，明确地表明了 妊娠期胎盘FSHR。目前研究的重点是子宫肌层FSHR在子宫肌层中的生理作用。 女性生育能力。我们发现FSHR在子宫肌层中有表达，在妊娠期间，它的表达显著 在学期中上调监管。值得注意的是，我们证明了卵泡刺激素促进了非妊娠妇女子宫肌层的静止。 当FSHR水平较低时，在妊娠早期，但在足月妊娠时，它刺激收缩活动 当FSHR水平较高时。虽然孕妇的脑下垂体促卵泡激素在怀孕期间受到抑制，但我们发表了 研究表明，FSH在胎盘、蜕膜和子宫肌层中合成，在那里它可以作为一种 旁分泌和/或自分泌信号与子宫肌层FSHR接触。我们假设并将在此验证 肌层FSHR在子宫肌层的建立和维持中起着重要的作用 怀孕的时间以及分娩的时间。具体地说，通过在早期促进子宫平静 我们假设，子宫肌层FSH/FSHR信号需要抑制子宫收缩和 从而促进胚胎着床，进而维持妊娠早期妊娠。在 妊娠末，当子宫肌层FSHR表达上调时，我们假设FSH/FSHR信号转导是 增加收缩活动所必需的，从而有助于分娩的时机。作为一个探索者 R21旨在提供子宫肌层FSHR有助于生育和成功 怀孕完成后，我们将利用一种新颖而强大的小鼠模型来验证我们的假设 使用CRISPR/Cas9插入FSHR外显子10两侧的loxP位点，以实现有条件的缺失 功能性FSHR来源于子宫肌层。如果研究结果支持我们的假设，结果将是范例 改变我们对FSH/FSHR信号在雌性生殖中的理解。它特别是 考虑到几项研究表明，植入失败、自然流产、 接受辅助生殖技术(ART)的不孕妇女的不良围产期结局是 不一定是由于ART，但可能是由于母体因素造成的不孕不育。如果其中一个因素是 卵巢FSHR对FSH的次优反应，由此得出子宫肌层类似的次优反应 FSHR到FSH可能导致无法获得或正常维持妊娠。评选结果 因此，拟议的研究可能会刺激基于子宫肌层的新治疗方法的发展 FSHR用于解决不孕症和不良妊娠结局。

PROJECT SUMMARY The role of the Follicle Stimulating Hormone Receptor (FSHR) in female fertility is thought to be restricted to the known actions of ovarian FSHR. Recently published studies from my laboratory challenge this dogma by demonstrating extra-ovarian FSHR expression in the female reproductive tract of cycling and pregnant women and the placenta and decidua of pregnant women, and unequivocally demonstrating a critical role for placental FSHR in pregnancy. The focus of the present studies is the physiological role of myometrial FSHR in female fertility. We show that FSHR is expressed in myometrium and that during pregnancy it is significantly up-regulated at term. Notably, we demonstrate that FSH promotes myometrial quiescence in the non-pregnant state and in early pregnancy when FSHR levels are low, but it stimulates contractile activity at term pregnancy when FSHR levels are high. Although maternal pituitary FSH is suppressed during pregnancy, our published studies suggest that FSH is synthesized in the placenta, decidua, and myometrium, where it can act as a paracrine and/or autocrine signal to engage the myometrial FSHR. We hypothesize and will test herein that myometrial FSHR contributes in a significant manner to both the establishment and the maintenance of pregnancy as well as the timing of parturition. Specifically, by promoting uterine quiescence early in pregnancy, we postulate that myometrial FSH/FSHR signaling is required to silence uterine contractions and thereby promote embryo implantation and the subsequent maintenance of pregnancy early in gestation. At the end of gestation, when myometrial FSHR expression is up-regulated, we postulate that FSH/FSHR signaling is required for increased contractile activity and thereby contributes to the timing of parturition. As an exploratory R21 designed to provide proof-of-principle that myometrial FSHR contributes to fertility and the successful completion of pregnancy, we will test our hypothesis utilizing a novel and powerful mouse model in which CRISPR/Cas9 was used to insert loxP sites flanking exon 10 of the Fshr to enable the conditional deletion of functional FSHR from myometrium. If the findings support our hypothesis, the outcomes would be paradigm shifting with respect to our understanding of FSH/FSHR signaling in female reproduction. It is particularly relevant to consider that several studies suggest that the risks of failed implantation, spontaneous miscarriage, and adverse perinatal outcomes in infertile women undergoing Assisted Reproductive Technologies (ART) are not necessarily due to ART, but may be due to maternal factors underlying infertility. If one such factor is a suboptimal response of ovarian FSHR to FSH, it follows that a similarly suboptimal response of myometrial FSHR to FSH may contribute to an inability to achieve or properly sustain pregnancy. The results of the proposed studies may therefore spur the development of novel therapeutic approaches based on myometrial FSHR to address infertility and poor pregnancy outcomes.