Signaling mechanisms that modulate uterine 3D structure for pregnancy success

项目概要/摘要 尽管对胚胎早期发育的了解很多，但子宫的结构 早期胚胎发育的环境还不清楚。当时子宫环境很差 胚泡进入和附着的可能会对生长的胎儿的健康造成长期的有害影响。 胚胎，导致缺陷，如流产，胎盘功能不全，子宫内生长受限， 先兆子痫和早产。使用共焦成像结合3D图像分析，我们有 确定和量化小鼠子宫腔结构在准备植入中的动态变化。 当应用于孕酮信号传导缺陷的小鼠突变体时， 或WNT 5A信号传导缺陷的小鼠，具有已知的分子植入缺陷，该方法揭示了 着床时子宫结构的显著异常。 这项提议的目标是确定胚胎和孕酮驱动的机制， 引导子宫折叠，为胚胎植入和妊娠成功做准备。在目标1中， 确定胚胎本身如何影响子宫的3D结构。我们将根据时间进程确定 沿系膜-反系膜轴沿着褶皱形成的时间模式。我们将确定 胚胎需要作为一个物理对象或作为一个生物信号中心，以引起结构的变化， 子宫腔在Aim 2中，我们将检验孕激素影响子宫内膜容受性的假设 通过塑造子宫的三维结构。首先，我们将评估人类受试者的子宫内膜折叠， 在月经周期中，雌激素占优势的增殖期和孕激素占优势的分泌期。我们将 然后采用生理、超生理孕酮处理，并对小鼠模型进行超数排卵， 增加孕酮水平，以确定孕酮如何调节折叠。我们还将使用鼠标 孕酮信号传导缺陷的突变体，以确定孕酮是否调节发育性折叠或 在怀孕早期。目标3中的研究将通过以下方式确定孕酮是否影响子宫腔形状： 与WNT 5A信号通路相互作用。我们还将测试胚胎的异常定位， 异常轴排列，在异常结构的褶皱，解释了整个不良妊娠结局， 异常折叠突变体（超数排卵和WNT 5A信号传导缺陷的突变体）。 在这项建议中开发的方法将是至关重要的分析子宫结构在三个- 不同的免疫缺陷基因突变体，病理条件的尺寸，并将有助于揭示 新的分子和结构途径参与成功的植入。我的长期愿景 研究是确定新的子宫三维结构的基础上的机制，管理子宫内膜容受性， 目标是制定新办法，改善辅助生殖的生育结果， 荷尔蒙失调患者的临床情况。

PROJECT SUMMARY/ABSTRACT Although much is known about the embryo during early development, the structural uterine environment in which the early embryo develops is not well understood. A poor uterine environment at the time of blastocyst entry and attachment can cause long lasting detrimental effects on the health of the growing embryo, leading to defects such as miscarriage, placental insufficiency, intra-uterine growth restriction, preeclampsia and preterm birth. Using confocal imaging in combination with 3D image analysis we have identified and quantified dynamic changes in murine uterine luminal structure in preparation for implantation. When applied to mouse mutants deficient in progesterone signaling, mice with excess progesterone signaling or mice deficient in WNT5A signaling, with known molecular implantation defects, this approach reveals striking abnormalities in uterine structure at the time of implantation. The goal of this proposal is to determine, embryo and progesterone driven mechanisms that guide uterine folding in preparation for embryo implantation and pregnancy success. In Aim 1 we will determine how the embryo itself affects the 3D structure of the uterus. Using a time-course we will determine the temporal pattern of fold formation along the mesometrial-anti mesometrial axis. We will determine if the embryo is required as a physical object or as a biological signaling center to cause structural changes in the uterine lumen. In Aim2 we will test the hypothesis that progesterone influences receptivity of the endometrium by shaping uterine 3D structure. First we will assess endometrial folding in human subjects in both the estrogen dominant proliferative and progesterone dominant secretory phase of the menstrual cycle. We will then use physiological, supra-physiological progesterone treatment, and a mouse model of superovulation with increased progesterone levels, to determine how progesterone regulates folding. We will also use mouse mutants deficient in progesterone signaling to determine if progesterone regulates folding developmentally or during early pregnancy. Studies in Aim 3 will determine if progesterone affects uterine luminal shape by interacting with the WNT5A signaling pathway. We will also test if aberrant localization of embryos and aberrant axis alignment, in aberrantly structured folds, explains the entirety of poor pregnancy outcomes in aberrant folding mutants (superovulated and mutants deficient in WNT5A signaling). The methods developed in this proposal will be crucial to analyze the uterine structure in three- dimensions for different implantation-defective genetic mutants, pathological conditions, and will help uncover novel molecular and structural pathways involved in successful implantation. The long-term vision of my research is to identify novel uterine 3D structure based mechanisms that govern endometrial receptivity with the goal of developing new approaches to improve fertility outcomes for assisted reproduction and potential clinical situations for patients with hormonal disruptions.