Signaling mechanisms that modulate uterine 3D structure for pregnancy success

项目摘要/摘要 尽管人们对胚胎在早期发育过程中的了解很多，但结构性子宫 早期胚胎发育的环境还不是很清楚。当时糟糕的子宫环境 胚泡进入和附着会对正在生长的人的健康造成长期的有害影响 胚胎，导致流产、胎盘功能不全、宫内生长受限、 先兆子痫和早产。使用共焦成像与3D图像分析相结合，我们有 鉴定和量化小鼠子宫腔结构在植入准备过程中的动态变化。 当应用于孕酮信号缺乏的突变小鼠时，孕酮信号过剩的小鼠 或者Wnt5A信号缺陷的小鼠，有已知的分子植入缺陷，这种方法揭示了 植入时子宫结构明显异常。 这项提议的目标是确定胚胎和黄体酮驱动的机制 指导子宫折叠，为胚胎植入和妊娠成功做好准备。在目标1中，我们将 确定胚胎本身如何影响子宫的3D结构。使用时间进程，我们将确定 沿子宫系膜-子宫反系膜轴褶皱形成的时间模式。我们将确定是否 胚胎需要作为物理对象或生物信号中心来引起结构上的变化 子宫腔。在AIM2中，我们将检验孕激素影响子宫内膜接受性的假设 通过塑造子宫的3D结构。首先，我们将评估两种情况下受试者的子宫内膜折叠 月经周期中以雌激素为主的增殖期和以黄体酮为主的分泌期。我们会 然后使用生理性、超生理性黄体酮治疗，并用小鼠模型进行超数排卵 提高孕酮水平，以确定孕酮如何调节折叠。我们还将使用鼠标 缺乏孕酮信号以确定孕酮是否调节发育或折叠的突变体 在怀孕早期。AIM 3的研究将确定孕酮是否通过以下方式影响子宫腔形状 与Wnt5A信号通路相互作用。我们还将测试胚胎的异常定位和 异常的轴排列，在异常结构的褶皱中，解释了不良妊娠结局的全部 异常折叠突变体(超数排卵和Wnt5A信号缺失突变体)。 这项建议中开发的方法将对分析三种子宫结构至关重要- 不同植入的维度--有缺陷的基因突变，病理条件，并将有助于发现 成功植入所涉及的新的分子和结构途径。我的长期愿景是 研究是确定新的基于子宫3D结构的机制，通过 开发改善辅助生殖生育结局的新方法的目标和潜力 荷尔蒙紊乱患者的临床情况。

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.