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

展示子宫肌层 FSHR 对生育力影响的小鼠模型

• 批准号: 9223342
• 负责人: DEBORAH SEGALOFF
• 金额: $ 22.88万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-30 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9223342
• 关键词: Address; Affect; Assisted Reproductive Technology; Attenuated; Autocrine Communication; Birth; Bladder; CRISPR/Cas technology; Cells; Couples; Cyclic AMP; Data; Decidua; Defect; Development; Enterobacteria phage P1 Cre recombinase; Equilibrium; Estrogens; Exons; Female; Female infertility; Fertility; Fertility Rates; Fetus; Follicle Stimulating Hormone Receptor; Foundations; Gastrointestinal tract structure; Growth; Health; Human; Infertility; Inositol Phosphates; Laboratories; Light; LoxP-flanked allele; Mediating; Mus; Muscle Fibers; Myometrial; Outcome; Ovarian; Ovarian Follicle; Ovary; Paracrine Communication; Pathway interactions; Physiological; Pituitary Gland; Placenta; Pregnancy; Pregnancy Maintenance; Pregnancy Outcome; Pregnancy loss; Pregnant Women; Publishing; Receptor Signaling; Reproduction; Risk; Role; Site; Smooth Muscle; Spontaneous abortion; Testing; Uterine Contraction; Uterus; Visceral; base; design; failure Implantation; fetal; healthy pregnancy; implantation; insight; mouse model; myometrium; natural Blastocyst Implantation; novel; novel therapeutic intervention; novel therapeutics; perinatal outcomes; pregnant; receptor density; receptor expression; reproductive tract; response; telokin; translational study; uterine contractility

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.

项目摘要 卵泡刺激素受体（FSHR）在女性生育中的作用被认为是有限的 卵巢FSHR的已知作用。我的实验室最近发表的研究挑战了这一教条 通过证实卵巢外FSHR在周期性和妊娠期女性生殖道中的表达， 妇女和孕妇的胎盘和蜕膜，并明确表明， 妊娠期胎盘FSHR。目前研究的焦点是子宫肌层FSHR在子宫内膜异位症中的生理作用。 女性生育能力我们发现，FSHR在子宫肌层表达，在妊娠期间， 在足月时上调。值得注意的是，我们证明，FSH促进非妊娠子宫肌层静止 当FSHR水平较低时，在怀孕早期，但在足月妊娠时刺激收缩活动 当FSHR水平很高时。尽管母体垂体FSH在怀孕期间受到抑制，但我们发表的 研究表明，FSH在胎盘、蜕膜和子宫肌层中合成， 旁分泌和/或自分泌信号参与子宫肌层FSHR。我们假设并将在此测试， 子宫肌层FSHR以重要的方式有助于建立和维持 怀孕以及分娩时间。具体来说，通过促进子宫静止早期， 怀孕时，我们假设子宫肌层FSH/FSHR信号传导需要沉默子宫收缩， 从而促进胚胎着床和随后在妊娠早期维持妊娠。在 在妊娠末期，当子宫肌层FSHR表达上调时，我们推测FSH/FSHR信号转导是通过 这是增加收缩活动所必需的，从而有助于分娩的时机。作为探索性 R21旨在提供子宫肌层FSHR有助于生育力和成功生育的原理证明。 完成妊娠后，我们将利用一种新的强大的小鼠模型来测试我们的假设， CRISPR/Cas9用于插入Fshr的外显子10侧翼的loxP位点，以使得能够条件性缺失Fshr的外显子10。 功能性FSHR。如果研究结果支持我们的假设， 我们对女性生殖中FSH/FSHR信号的理解发生了变化。显得尤为 考虑到几项研究表明，植入失败，自发流产， 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，但可能是由于不孕症的母体因素。如果其中一个因素是 卵巢FSHR对FSH的次优反应，因此子宫肌层的类似次优反应 FSHR到FSH可能导致无法实现或适当维持妊娠。的结果 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解决不孕症和不良妊娠结局。