Maternal Uterine Vascular Origins of Fetal Alcohol Spectrum Disorders

成功预防或改善酒精致畸作用的努力受到了阻碍，至少 部分原因是对酒精损害发育中的胎儿的机制了解有限。在 在K99/R 00基金中，我们将探索胎儿酒精谱系障碍（FASD）的母体子宫起源 并设计一种策略，用于开发未来的蛋白质组学生物标志物/独特的特征谱， 母亲饮酒。整个子宫循环的协调生长和重塑， 胎盘的形成是胎儿正常发育的基础。这些复杂的过程是由 通过内皮源性一氧化氮（NO）和内皮型一氧化氮合酶（eNOS）的酶活性。的 这项提案的总体目标是调查慢性酗酒对以下方面的直接影响：1）NO， 妊娠期间子宫动脉内皮中eNOS相关的信号级联;和2）小窝， eNOS的天然家园，并利用这一知识开发高通量蛋白质组学 母亲饮酒的生物标志物/独特特征谱，NIAAA战略的既定目标 2009-2014年计划。独特的途径调节NO和eNOS在怀孕的子宫，这些发挥作用， 在妊娠相关的母体子宫血管适应中的独特作用。在具体目标#1中，我们将 直接比较酗酒介导的适应性反应和特定的信号通路， 妊娠子宫动脉内皮细胞在剪切应力下通过分级脉动体内样流动条件。 从这些研究中获得的数据将为理解 剪切应力和酒精的相互作用，以调节NO的生产在怀孕的子宫内皮。 酗酒改变了eNOS和cav-1之间的化学计量关系， [Ca+2]i显著升高，eNOS被驱离其“天然家园”小窝， 这是一个主要的稳定环境。在具体目标#2中，我们将研究酒精诱导的 细胞内[Ca+2]i的反复增加及其对细胞膜小窝中eNOS反复耗竭的影响， 没有生产。在具体的目标#3中，我们将利用高通量蛋白质组学来鉴定一种 生物标志物/独特的小窝标记蛋白质谱，其依赖于酒精损伤的水平。这些 研究结果将使我们处于一个很好的位置，以了解酒精损害的多机制原因， 特别是从母亲和子宫的角度，并正确设计和提出一个 全面的预防策略。

Efforts to successfully prevent or ameliorate the teratogenic effects of alcohol have been impeded, at least in part, by a limited understanding of the mechanisms by which alcohol damages the developing fetus. In this K99/R00 grant, we will explore the maternal uterine origins of Fetal Alcohol Spectrum Disorders (FASD) and devise a strategy for development of a future proteomic biomarker(s)/unique signature profile for maternal alcohol consumption. Coordinated growth and remodeling of the entire uterine circulation and creation of a placenta are requisites for normal fetal development. These intricate processes are controlled by endothelial-derived nitric oxide (NO) and enzyme activity of endothelial nitric oxide synthase (eNOS). The overall goal of this proposal is to investigate the direct effects of chronic binge alcohol on: 1) NO and eNOS-related signaling cascades in the uterine artery endothelium during pregnancy; and 2) the caveolae, the natural home for eNOS, and to utilize this knowledge to develop a high throughput proteomic biomarker(s)/unique signature profile for maternal alcohol consumption, a stated goal of NIAAA strategic plan for years 2009-2014. Unique pathways regulate NO and eNOS in the pregnant uterus and these play a distinct role in pregnancy-associated maternal uterine vascular adaptations. In specific aim#1, we will directly compare binge alcohol mediated adaptive responses and specific signaling pathways in the pregnant uterine artery endothelial cells under shear stress via graded pulsatile in vivo-like flow conditions. Data derived from these studies will provide the first mechanistic framework for understanding the interactions between shear stress and alcohol to regulate NO production in pregnant uterine endothelium. Binge alcohol alters the stoichiometric relationship between eNOS and cav-1 and with every bout of alcohol, there are significant rises in [Ca+2]i and in turn eNOS is driven away from the caveolae, its "natural home" which acts as a major stabilizing environment. In specific aim #2, we will investigate alcohol-induced repeated intracellular increases in [Ca+2]i and its effects on repeated depletion of eNOS from caveolae and NO production. In specific aim #3, we will utilize high throughput proteomics to identify a biomarker(s)/unique caveolar signature protein profile that is dependent on the level of alcohol insult. These findings will place us in an excellent position to understand the multimechanistic causes of alcohol damage, especially from the perspective of the mother and the uterus, and to correctly design and propose a comprehensive preventative strategy.