A Novel Mechanistic Framework for FASD Etiology.

FASD 病因学的新机制框架。

基本信息

批准号：
10459965
负责人：
Jayanth Ramadoss
金额：
$ 36.29万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-15 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10459965
关键词：
3-Dimensional Affect Agonist Alcohol abuse Alcohol consumption Alcohols Animal Model Arteries Arteriographies Behavioral Biological Availability Biological Markers Blood Vessels Blood flow Brain Cephalic Cerebrum Child Complex Data Development Disease model Electrophysiology (science)Endothelium Ethanol Metabolism Etiology FRAP1 gene Fetal Alcohol Exposure Fetal Alcohol Spectrum Disorder Fetal Growth Fetal Growth Retardation Functional disorder Future Grant Growth Growth Disorders High Pressure Liquid Chromatography Image Immunoblotting Immunofluorescence Immunologic Impairment In Vitro Individual Learning Disabilities Lecithin Measures Mediating Mediator of activation protein Microspheres Modeling Molecular Molecular Target Morphology Myography NOS3 gene Neurobiology Neurodevelopmental Deficit Neurons Nitric Oxide Nutrient Organ Outcome Oxygen Pathway interactions Pharmacology Phenotype Phosphatidic Acid Physiological Play Positioning Attribute Pregnancy Prevention Preventive Production Psyche structure Rattus Reporting Reverse Transcriptase Polymerase Chain Reaction Role School-Age Population Signal Transduction Societies System Teratogenic effects Tissues Toothache Ultrasonography Uterus Vasodilation alcohol consumption during pregnancy alcohol exposure ascending aorta basilar artery body system cost cost estimate design disability experimental study fetal human model improved in vivo indexing insight maternal alcohol use middle cerebral artery neuron development novel patch clamp phosphatidylethanol pregnant pressure prevent therapy design transcriptome sequencing

项目摘要

Fetal Alcohol Spectrum Disorders (FASD) affects an estimated 2–5% of young school age children in the U.S., with an estimated cost of $1.4 million per individual. Two cardinal outcomes of FASD are fetal growth restriction and neurodevelopmental deficits. Efforts to successfully prevent or ameliorate these teratogenic effects of alcohol abuse have been impeded by a limited understanding of alcohol’s complex mechanisms of action, which impact multiple organ systems. In addition, etiological reports on FASD outcomes have been mostly limited to investigating indirect downstream mediators. We propose the molecular pathway governing phosphatidylethanol (PEth; 100% specific, most sensitive biomarker for gestational alcohol exposure) formation can yield novel insights into FASD etiology, as during alcohol metabolism, phosphatidylcholine is hydrolyzed to PEth instead of phosphatidic acid (PA, an essential nutrient for growth/neuron development). In an established FASD model, our novel preliminary data shows alcohol decreases PA bioavailability and concurrently increases PEth levels in maternal and fetal compartments. Our data also show alcohol-induced impairment of maternal uterine artery (related to fetal growth) and fetal brain vascular (related to neurodevelopmental outcomes) adaptations. Interestingly, PA addition in vitro to the uterine and middle cerebral arteries reverses alcohol-induced dysfunction in these vessels, and in vivo PA administration reverses FASD growth deficit. Our data also identify a role for endothelial nitric oxide (NO) synthase (eNOS) and mTORC1 signaling in this alcohol/PEth/PA framework. In Aim #1, we hypothesize that in our FASD model, PA plays a major role in alcohol-mediated vasodilatory deficits and the related eNOS pathway in maternal uterine and developing cranially directed arteries, and that alcohol impairs the NO system via PA-mediated mTORC1 system alteration. Following mechanistic in vitro blockade of PA, mTORC1, and related signaling, we will assess uterine and developing cranially directed arterial adaptations using arteriography, LC-MS/MS, immunoblotting, immunofluorescence, RNA-seq, RT-PCR, and patch clamp. In Aim#2, we hypothesize PA administration in vivo reverses alcohol-induced decreases in uterine artery and fetal cranially directed blood flow, and improves fetal nutrient delivery, growth phenotypes, and deficits in alcohol-sensitive neurobiological outcomes. We will measure growth indices, uterine blood flow, uterine O2/nutrient delivery, fetal cranially directed blood flow, and neuronal function/morphology to assess the role of PA in the etiology of two cardinal FASD outcomes. We anticipate that the proposed experiments will provide a much-needed breakthrough in the FASD field by identifying a promising etiological molecular pathway(s) for FASD growth and/or neurodevelopmental deficits. These studies will pave way for future novel prevention/treatment studies strategically aimed at rescuing FASD cardinal outcome phenotypes through manipulation of direct alcohol targets.

胎儿酒精谱系障碍 (FASD) 影响着美国约 2-5% 的学龄儿童，估计每人损失 140 万美元，胎儿生长受限和神经发育缺陷是 FASD 的两个主要后果。对酒精复杂作用机制的了解有限，阻碍了对酒精滥用的这些致畸作用的研究或改善，酒精影响多个器官系统。此外，关于酒精的病因学报告。 FASD 结果主要局限于研究间接下游介质，我们认为控制磷脂酰乙醇（PEth；妊娠期酒精暴露的 100% 特异性、最敏感的生物标志物）形成的分子途径可以对 FASD 病因学产生新的见解。在酒精代谢过程中，磷脂酰胆碱被水解为 PEth 而不是磷脂酸（PA，生长/神经元发育的必需营养素）。在已建立的 FASD 模型中，我们的新初步数据显示，酒精会降低 PA 生物利用度，同时增加母体和胎儿的 PEth 水平。我们的数据还表明，酒精会导致母体子宫动脉（与胎儿生长相关）和胎儿脑血管（与神经发育结果相关）适应受损。体外添加到子宫和大脑中动脉可逆转酒精引起的这些血管功能障碍，体内 PA 给药可逆转 FASD 生长缺陷，我们的数据还确定了内皮一氧化氮 (NO) 合酶 (eNOS) 和 mTORC1 信号在这些血管中的作用。在目标 1 中，我们研究了酒精/PEth/PA 框架，在我们的 FASD 模型中，PA 在酒精介导的血管舒张缺陷和母体相关的 eNOS 通路中发挥着重要作用。子宫和颅内定向动脉，并且酒精通过 PA 介导的 mTORC1 系统改变损害 NO 系统，在体外机械阻断 PA、mTORC1 和相关信号传导后，我们将开发评估子宫和颅内定向动脉。使用动脉造影、LC-MS/MS、免疫印迹、免疫荧光、RNA-seq、RT-PCR 和膜片钳进行动脉适应在 Aim#2 中，我们探索了体内 PA 给药可逆转酒精引起的子宫动脉和胎儿颅骨定向减少。血流量，并改善胎儿营养输送、生长表型和酒精敏感神经生物学结果的缺陷。我们将测量生长指数、子宫血流量、子宫氧气/营养输送、胎儿。我们预计，所提出的实验将通过确定一个有前景的病因分子途径，为 FASD 领域提供急需的突破。 s) 用于 FASD 生长和/或神经发育这些研究将为未来的新型预防/治疗研究铺平道路，这些研究的战略目标是通过操纵直接酒精目标来挽救 FASD 的主要结果表型。