Dissecting gene dysregulation at the maternal-fetal interface in preeclampsia

项目总结/摘要 我们正在验证一个假设，即主要滋养层细胞（TB）的整体基因表达谱 来自先兆子痫（PE）患者的胎盘亚群将导致新分子的鉴定 在这种综合症中起着重要作用。这一战略的动力是发现PE始终是 与某些胎盘病变有关细胞滋养层（CTB）浸润的间质成分是 通常局限于表层蜕膜。同样，血管内侵入也受到限制， 涉及的螺旋小动脉的数量和它们被修改的程度。合体滋养细胞 感染患者胎盘中的STBs也有明显的变化，如合胞结。这些改变 伴随着分子变化，其中许多已被确定为分析，在PE，表达 这些分子通常在结核病的功能中起着重要作用。但是，我们已经开始申请 公正的方法可以更全面地了解结核病基因表达的变化， 发生在PE这种方法承认我们对胎盘成分的了解有限， 综合征最近，我们使用转录组学的方法来分析CTB基因在各种严重肿瘤中的表达， PE（sPE）。具体而言，从sPE患者和对照女性的胎盘中分离的CTB 培养48小时以允许分化/侵袭。微阵列分析显示sPE相关 一组CTB基因的上调，到培养期结束时，恢复到对照水平。他们 包括以前与PE相关的因子和许多新的分子，包括血管生成调节因子， SEMA3B。我们继续表明，这种神经纤毛蛋白-1和-2配体的水平升高， PE对CTB的作用机制包括通过PI 3 K/AKT下调VEGF信号传导， GSK 3通路。在sPE CTB中观察到相同的变化。我们的数据支持这样的理论，在体育课上， 体内环境损害CTB分化/侵袭，差异表达的分子有助于 的机制，并认为临床体征是由患者的具体因素。在确定了一个 由于sPE中侵袭性CTB的基因特征，我们现在提出了对其他三种受影响的CTB的无偏分析。 TB人群、STB、合胞结和血管内CTB。具体来说，我们将使用激光显微切割 从PE、sPE和对照胎盘中分离这些细胞， 确定失调基因（目标1）。我们将使用我们新的细胞培养模型，人类结核祖细胞， 确定观察到的变化的功能意义（目的2）。我们认为这些实验 提供了关于PE中胎盘缺陷的分子基础的新信息。这些发现也可能 显著的平移潜力，例如，失调分子的一个子集可能是循环生物标志物， 和/或改善胎盘功能的治疗靶点，我们的数据表明这是可能的。

Project Summary/Abstract We are testing the hypothesis that global gene expression profiling of the major trophoblast (TB) subpopulations of placentas from preeclampsia (PE) patients will lead to the identification of novel molecules that play important roles in this syndrome. The impetus for this strategy is the finding that PE is consistently associated with certain placental pathologies. The interstitial component of cytotrophoblast (CTB) invasion is frequently restricted to the superficial decidua. Likewise, endovascular invasion is constrained in terms of the number of spiral arterioles that are involved and the extent to which they are modified. Syncytiotrophoblasts (STBs) from placentas of affected patients also have overt changes such as syncytial knots. These alterations are accompanied by molecular changes, many of which have been identified by profiling, in PE, the expression of molecules that normally play important roles in terms of TB functions. However, we have begun to apply unbiased approaches to gain a more comprehensive understanding of changes in TB gene expression that occur in PE. This approach acknowledges our limited understanding of the placental component of this syndrome. Recently, we used a transcriptomics approach to profile CTB gene expression in various severe forms of PE (sPE). Specifically, CTBs that were isolated from the placentas of sPE patients and control women were cultured for 48 h to allow differentiation/invasion. Microarray analyses revealed sPE-associated upregulation of a suite of CTB genes that, by the end of the culture period, returned to control levels. They included factors previously associated with PE and many novel molecules, including the angiogenic regulator, SEMA3B. We went on to show that elevating levels of this neuropilin-1 and -2 ligand phenocopied many of the effects of PE on CTBs by mechanisms that include downregulating VEGF signaling through the PI3K/AKT and GSK3¿/¿ pathway. The same changes were observed in sPE CTBs. Our data support the theory that, in PE, the in vivo environment impairs CTB differentiation/invasion, the differentially expressed molecules contribute to the mechanisms, and that the clinical signs are determined by patient-specific factors. Having identified a genetic signature for invasive CTBs in sPE, we now propose an unbiased analysis of the three other affected TB populations, STBs, syncytial knots and endovascular CTBs. Specifically, we will use laser microdissection to isolate these cells from PE, sPE and control placentas and a global transcriptional profiling approach to identify the dysregulated genes (Aim 1). We will employ our new cell culture model, human TB progenitor cells, to determine the functional significance of the observed changes (Aim 2). We think that these experiments will yield new information about the molecular bases of placental defects in PE. The findings could also have significant translational potential, e.g., a subset of the dysregulated molecules could be circulating biomarkers and/or therapeutic targets for improving placental function, which our data suggest is possible.