Maternal Uterine Vascular Origins of Fetal Alcohol Spectrum Disorders

描述（由申请人提供）：成功预防或改善酒精致畸作用的努力受到阻碍，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。在这项K99/R00资助中，我们将探索胎儿酒精谱系障碍（FASD）的母体子宫起源，并制定一项策略，用于开发未来的蛋白质组学生物标志物/母亲饮酒的独特特征谱。整个子宫循环的协调生长和重塑以及胎盘的形成是胎儿正常发育的必要条件。这些复杂的过程是由内皮来源的一氧化氮（NO）和内皮一氧化氮合酶（eNOS）的酶活性控制的。本应用程序的总体目标是研究慢性酗酒对妊娠期间子宫动脉内皮NO和enos相关信号级联的直接影响；2)小泡，eNOS的天然家园，并利用这些知识开发高通量蛋白质组学生物标志物/产妇饮酒的独特特征谱，这是NIAAA 2009-2014年战略计划的既定目标。在妊娠相关的母体子宫血管适应中，有独特的途径调节NO和eNOS。在具体目标#1中，我们将通过分级脉动在体内样血流条件下直接比较酗酒介导的子宫动脉内皮细胞在剪切应力下的适应性反应和特定信号通路。来自这些研究的数据将为理解剪切应力和酒精之间调节妊娠子宫内皮NO生成的相互作用提供第一个机制框架。狂饮酒精会改变eNOS和cav-1之间的化学计量关系，每喝一次酒，[Ca+2]i就会显著升高，反过来eNOS就会被赶出作为主要稳定环境的“天然家园”——小泡。在特定目标#2中，我们将研究酒精诱导的细胞内[Ca+2]i的重复增加及其对小泡中eNOS的重复消耗和NO生成的影响。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/独特的空洞特征蛋白谱。这些发现将使我们更好地理解酒精损伤的多机制原因，特别是从母亲和子宫的角度，并正确设计和提出全面的预防策略。

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.