A Novel Mechanistic Framework for FASD Etiology.

FASD 病因学的新机制框架。

• 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
• 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

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（FASD）估计有2-5％的年轻学龄儿童，估计每人成本为140万美元。 FASD的两个主要结果是胎儿生长限制和神经发育缺陷。对酒精对饮酒的复杂作用机理的了解有限，这会成功预防或改善酗酒的这些致力作用，从而阻碍了多器官系统。此外，关于FASD结局的病因报告主要仅限于研究间接下游介体。我们提出了控制磷脂酰乙醇的分子途径（Peth;特异性，最灵敏的生物标志物用于妊娠酒精暴露）形成可以产生对FASD病因的新见解，因为 在酒精代谢过程中，将磷脂酰胆碱水解为Peth而不是磷脂酸（PA，一种生长/神经元发展的必需营养素）。在既定的FASD模型中，我们的新型初步数据显示，酒精可降低生物利用度，并同时提高母体和胎儿室中的Peth水平。我们的数据还表明，酒精引起的母体子宫动脉（与胎儿生长有关）和胎儿脑血管（与神经发育结局有关）的损伤。有趣的是，PA在体外添加到子宫和大脑中动脉中，逆向酒精引起的这些血管中的功能障碍，并且在体内给药会逆转FASD的生长不足。我们的数据还确定了该酒精/Peth/PA框架中内皮一氧化氮（NO）合酶（ENOS）和MTORC1信号的作用。在AIM＃1中，我们假设在FASD模型中，PA在醇介导的血管舒张中起着重要作用，定义了材料子宫和颅面定向的动脉的相关eNOS途径，并且酒精通过PA介导的MTORC1系统改变了NO系统。遵循机械的体外阻断PA，MTORC1和相关信号传导，我们将评估子宫并发展为颅骨的定向 使用动脉造影，LC-MS/MS，免疫印迹，免疫荧光，RNA-SEQ，RT-PCR和斑块夹的动脉适应。在AIM＃2中，我们假设PA在体内给药可逆转酒精引起的子宫动脉和胎儿颅骨定向的血液流量下降，并改善胎儿营养的递送，增长的表型，并在酒精敏感的神经生物学成果中定义。我们将测量生长指数，子宫血流，子宫O2/营养递送，胎儿颅骨定向的血流以及神经发育，以评估PA在两个基本FASD结果的病因中的作用。我们预计，提出的实验将通过识别有希望的病因学分子途径来为FASD生长和/或神经发育提供有希望的病因学分子途径，从而在FASD领域提供急需的突破。 缺陷。这些研究将为未来的新型预防/治疗研究铺平道路，旨在通过操纵直接算法靶标来挽救FASD基本结果表型。