Maternal Uterine Vascular Origins of Fetal Alcohol Spectrum Disorders

描述（由申请人提供）：由于对酒精损害发育中胎儿的机制了解有限，成功预防或改善酒精致畸作用的努力至少部分受到阻碍。在这项K99/R 00资助中，我们将探索胎儿酒精谱系障碍（FASD）的母体子宫起源，并制定一项战略，用于开发未来的蛋白质组生物标志物/母亲饮酒的独特特征。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所必需的。这些复杂的过程由内皮源性一氧化氮（NO）和内皮型一氧化氮合酶（eNOS）的酶活性控制。本申请的总体目标是研究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中的NO和eNOS相关信号级联;和2）小窝，eNOS的天然家园，并利用这一知识来开发母亲饮酒的高通量蛋白质组学生物标志物/独特特征谱，2009-2014年NIAAA战略计划的既定目标。独特的途径调节NO和eNOS在妊娠子宫和这些发挥独特的作用，妊娠相关的母体子宫血管适应。在具体的aim#1中，我们将直接比较在剪切应力下通过分级脉动体内样流动条件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信号传导途径。从这些研究中获得的数据将提供第一个机制框架，了解剪切应力和酒精之间的相互作用，以调节NO的生产在怀孕的子宫内皮细胞。酗酒改变了eNOS和cav-1之间的化学计量关系，并且随着每次饮酒，[Ca+2]i显著升高，进而eNOS被驱离小窝，其“天然家园”充当主要的稳定环境。在具体目标#2中，我们将研究酒精诱导的[Ca+2]i的重复细胞内增加及其对来自小窝的eNOS重复消耗和NO产生的影响。在具体目标#3中，我们将利用高通量蛋白质组学来鉴定依赖于酒精损伤水平的生物标志物/独特的小窝特征蛋白质谱。这些发现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。 公共卫生相关性：在美国，每年至少有40，000名婴儿出生时患有FASD，估计每人花费140万美元，总成本至少为60亿美元。成功预防或改善酒精致畸作用的努力受到阻碍，至少部分是由于对酒精损害发育中胎儿的机制的了解有限。在这项K99/R 00资助中，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，用于开发高通量蛋白质组生物标志物/母亲饮酒的独特特征。

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.