Maternal Uterine Vascular Origins of Fetal Alcohol Spectrum Disorders

描述(由申请人提供)：至少在一定程度上，由于对酒精损害发育中胎儿的机制的有限了解，成功预防或改善酒精致畸作用的努力受到了阻碍。在这笔K99/R00赠款中，我们将探索胎儿酒精谱系障碍(FASD)的母体子宫起源，并制定一项策略，以开发一种未来的蛋白质组生物标记物(S)/母体饮酒的独特特征图谱。整个子宫循环的协调生长和重塑以及胎盘的形成是正常胎儿发育的必要条件。这些复杂的过程受内皮源性一氧化氮(NO)和内皮型一氧化氮合酶(ENOS)活性的控制。这项应用的总体目标是调查慢性酗酒对以下方面的直接影响：1)妊娠期间子宫动脉内皮细胞中NO和eNOS相关的信号级联；以及2)eNOS的天然家园--小窝，并利用这一知识开发高通量蛋白质组生物标记物(S)/母亲饮酒的独特特征图谱，这是美国国家酒类协会战略计划2009年至2014年的明确目标。独特的途径调节怀孕子宫中的NO和eNOS，它们在妊娠相关的母体子宫血管适应中发挥着不同的作用。在特定的目标#1中，我们将通过分级脉动的活体血流条件，直接比较在切应力下妊娠子宫动脉内皮细胞中酗酒介导的适应性反应和特定的信号通路。来自这些研究的数据将提供第一个机制框架，以了解切应力和酒精之间的相互作用，以调节怀孕子宫内皮NO的产生。酗酒改变了eNOS和Cav-1之间的化学计量关系，每喝一次酒，[Ca+2]i就会显著上升，反过来eNOS会被赶出小窝，小窝是eNOS的“天然家园”，起着主要稳定环境的作用。在特定的目标#2中，我们将研究酒精引起的细胞内[Ca+2]i的反复增加及其对小窝eNOS和NO产生的反复耗竭的影响。在具体目标#3中，我们将利用高通量蛋白质组学来识别依赖于酒精侮辱水平的生物标记物(S)/独特的凹陷特征蛋白图谱。这些发现将使我们处于一个很好的位置来理解酒精损害的多机制原因，特别是从母亲和子宫的角度，并正确地设计和提出全面的预防策略。 与公共健康相关：在美国，每年至少有4万名婴儿出生时患有FASD，估计每个人的成本为140万美元，总成本至少为60亿美元。至少在一定程度上，由于对酒精损害发育中胎儿的机制的有限了解，成功地预防或改善酒精的致畸作用的努力受到了阻碍。在这笔K99/R00赠款中，我们将探索胎儿酒精谱系障碍(FASD)的母体子宫起源，并制定一项策略，以开发高通量蛋白质组生物标记物(S)/母体饮酒的独特特征图谱。

DESCRIPTION (provided by applicant): 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 application 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. PUBLIC HEALTH RELEVANCE: Each year, at least 40,000 babies are born with FASD in the U.S. at an estimated cost of $1.4 million per individual and total cost of at least $6 billion. 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 a high throughput proteomic biomarker(s)/unique signature profile for maternal alcohol consumption.