Evaluating role of complement activation induced signaling pathways in preeclampsia pathology using a novel complement activation-based mouse model

子痫前期(PE)是一种妊娠特有的综合征，几种可能的机制被认为与 PE的发病机制包括补体(C)级联激活。循环中的激活副产物 在怀孕妇女中升高，在PE妇女中进一步显着升高。胎盘 与血压正常的女性相比，PE患者体内C活化副产物的沉积也显著增加。这个 胎儿/胎盘的半同种异体性质诱导母体C级联激活。为了了解原因-和- C活化与妊娠高血压综合征的作用关系 胎盘C活化与妊娠高血压综合征和胎儿生长受限的机制 将这些通路作为潜在治疗发现的目标的观点。一种可能的机制是C 激活可能促进全身性抗血管生成和促炎环境，母体血管功能障碍， 高血压和肾脏病理改变。此外，C的激活可能会诱导胎盘纤维蛋白沉积，这是由于 串音配合凝血系统，促进胎儿生长受限。在此应用程序中，我们将确定 用一种新的方法研究C激活与妊娠高血压综合征和胎儿生长受限的关系 补体激活为主的小鼠模型。这种转基因小鼠模型允许我们下调Crry在 一种可诱导的、有条件的和特定的胎盘方式，并研究其对各种母体系统的影响。 具体目的1.评估胎盘特异性C激活是否促进全身性抗血管生成 和促炎环境、高血压、母体血管功能障碍和肾脏病理 通过C3a-C3aR和/或C5a-C5aR、STAT3和ERK通路。假说：C蛋白在 怀孕促进全身抗血管生成(sFlt1、seng)和促炎水平升高 分子(肿瘤坏死因子-α、IL-6和IL-8)，降低松弛和增加收缩反应(血管 母体外周动脉功能障碍)、高血压、子宫螺旋动脉(SA)缺陷 通过C3a-C3aR/C5a-C5aR、STAT3和 Erk小路。此外，使用特定的抑制剂CR2-CRRY抑制C活化可改善这些C 引发的变化。具体目标2.检验C激活诱导胎盘纤维蛋白的假说 通过与凝血系统的串扰沉积促进胎儿生长受限。 假设：C活化通过减少胎儿血液供应促进胎儿生长受限 组织因子和von Willebrand对胎盘绒毛周围纤维蛋白沉积的破坏作用 因子(VWF)动态平衡。C直接通过C5a-C5aR信号激活，间接通过 促炎细胞因子IL-6、IL-8和肿瘤坏死因子-α诱导纤溶酶原激活剂TF上调 抑制物1和vWF通过内皮细胞，下调肝脏中ADAMTS-13的表达。

Preeclampsia (PE) is a pregnancy-specific syndrome and several putative mechanisms have been implicated in the pathogenesis of PE including complement (C) cascade activation. C activation byproducts in the circulation are elevated with pregnancy in women and they are further elevated significantly in PE women. Placental deposition of C activation byproducts is also significantly elevated in PE compared to normotensive women. The semi-allogenic nature of fetus/placenta induces maternal C cascade activation. To understand the cause-and- effect relation between C activation and pregnancy hypertension, it is essential to gain insight into the mechanistic pathways that link placental C activation to pregnancy hypertension and fetal growth restriction with the view of targeting these pathways for potential therapeutic discoveries. One possible mechanism is that C activation may promote systemic antiangiogenic and proinflammatory milieu, maternal vascular dysfunction, hypertension and renal pathological changes. Further, C activation may induce placental fibrin deposition due to crosstalk with coagulation system, promoting fetal growth restriction. In this application we will determine the mechanisms that link C activation to pregnancy hypertension and fetal growth restriction using a novel complement activation-based mouse model. This transgenic mouse model allows us to downregulate Crry in an inducible, conditional and placenta specific manner and study its effects on various maternal systems. Specific aim 1. To assess if placenta specific C activation promotes systemic antiangiogenic and proinflammatory milieu, hypertension, maternal vascular dysfunction, and renal pathology through C3a-C3aR and/or C5a-C5aR, STAT3 and ERK pathways. Hypothesis: C activation during pregnancy promotes increased systemic levels of antiangiogenic (sFLT1, sEng) and proinflammatory molecules (TNF-α, IL-6 and IL-8), reduced relaxation and increased contractile responses (vascular dysfunction) of maternal peripheral arteries, hypertension, defective uterine spiral artery (SA) remodeling, and glomerular endotheliosis (renal pathology) through C3a-C3aR/C5a-C5aR, STAT3 and ERK pathways. Further, inhibition of C activation using specific inhibitor CR2-Crry ameliorates these C induced changes. Specific aim 2. To test the hypothesis that C activation induces placental fibrin deposition through its crosstalk with coagulation system promoting fetal growth restriction. Hypothesis: C activation promotes fetal growth restriction by reducing fetal blood supply due to placental peri-villous fibrin deposition through the disruption of tissue factor (TF) and von Willebrand factor (vWF) homeostasis. C activation directly through C5a-C5aR signaling and indirectly through proinflammatory cytokines IL6, IL8 and TNF-α induces the upregulation of TF, plasminogen activator inhibitor 1 and vWF by endothelial cells and downregulates the ADAMTS-13 in liver.