Acute Prenatal Alcohol Exposure Potentiates Conotruncal Defects in the Setting of a Permissive Genetic Background

在允许的遗传背景下，急性产前酒精暴露会加剧圆锥干缺陷

• 批准号: 10731366
• 负责人: Drayton C Harvey
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-18 至 2027-05-17
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10731366
• 关键词: Accidents; Acetylation; Acids; Acute; Address; Alcohol consumption; Alcohols; Apoptosis; Automobile Driving; Awareness; Biological Assay; Biology; Cardiac; Cementation; Chest; Childhood; Chronic; Clinical; Clinical Research; Collaborations; Congenital Abnormality; Congenital Heart Defects; Craniofacial Abnormalities; Critical Pathways; Data; Defect; Development; Diagnosis; Doctor of Medicine; Doctor of Philosophy; Education; Ensure; Ethanol; Exposure to; Fellowship; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Fetal alcohol effects; Frequencies; Genes; Genetic; Genetic Transcription; Heart; Histologic; Histones; Hospitalization; In Vitro; Incidence; Individual; Institution; Intervention; Intraperitoneal Injections; Investigation; Ligands; Literature; Lysine; Mentorship; Molecular; Mus; Mutation; National Institute on Alcohol Abuse and Alcoholism; Nature; Neonatal Mortality; Neurologic; Organ; Organ Culture Techniques; Organogenesis; Participant; Pathway interactions; Patients; Population; Pregnancy; Prevention; Principal Investigator; Proliferating; Regression Analysis; Relative Risks; Reporting; Research; Reverse Transcription; Risk; Risk Factors; Role; Sampling; Severities; Signal Transduction; Site; Specific qualifier value; Specificity; Teratogens; Testing; Therapeutic Intervention; Time; Training; Validation; Variant; Work; alcohol effect; alcohol exposure; alcohol risk; body system; cardiogenesis; clinically relevant; conotruncal heart defect; cost; disorder prevention; drinking; epigenetic regulation; genome sequencing; histone acetyltransferase; human study; in vivo; inhibitor; malformation; migration; mouse model; notch protein; novel; overexpression; permissiveness; prevent; preventive intervention; progenitor; programs; restoration; synergism; whole genome

PROJECT SUMMARY/ABSTRACT Nearly 30% of patients with Fetal Alcohol Spectrum Disorder (FASD) have a congenital heart defect (CHD). Outflow tract (OFT) defects are over-represented, though with variable incidence and severity. The majority of prenatal alcohol exposure (PAE) research focuses on the hallmarks of FASD, chronic drinking and neurologic defects. The most common form of PAE in the population, acute exposure during the periconceptual period, and its effect on cardiac development have not been studied. We will study the effects of acute, periconceptual drinking both through a multi-institutional clinical study and with our acute PAE murine model, in which mice receive two intraperitoneal injections of 3g/kg of 30% ethanol at a point critical to cardiac organogenesis. We hypothesize the variable incidence and severity of PAE-induced OFT defects can be explained by a combination of PAE and otherwise non-deleterious mutations that result in a genetically permissive background. We believe mutations in the Notch pathway establish such a genetically permissive background, and that PAE acts synergistically with these mutations via epigenetic regulation of Notch to result in OFT malformation. Aim 1 of this study will address the specificity of the deleterious effects of PAE to OFT alignment and define the molecular pathways disrupted in individuals with PAE-induced OFT defects. We will achieve this through relative risk analysis of PAE and CHD diagnoses and pathway and subsequent logistical regression analyses of whole genome sequencing data. Aim 2 will define alcohol driven epigenetic regulation as the mechanism by which acute PAE and otherwise non-deleterious Notch pathway mutations synergistically disrupt OFT development. Using a combination of molecular assays and histologic analysis in vitro and in vivo, we will test the impact of this combined teratogenic insult on second heart field (SHF) viability and ability to migrate into the OFT. We will establish PAE driven epigenetic regulation, disrupting Notch gene accessibility and transcription, inhibits Notch signaling and use a pan-histone acetyltransferase inhibitor in vitro to demonstrate prevention of PAE induced hyperacetylation is sufficient to rescue Notch signaling and SHF viability. Similarly, rescue of SHF viability by overexpression of notch intracellular domain (NICD) in vitro will cement the point of acute PAE and Notch mutation synergy as loss of Notch signaling prohibiting SHF viability. As one of the first studies to examine the interaction of clinically relevant acute PAE with heart development, this study addresses the priorities of the NIAAA to define the impact of non-chronic exposure and alcohol’s effects on understudied organ systems. The genetic pathways identified by this study, including novel validation of the Notch pathway as significant in PAE-induced CHD, will provide targets for disease prevention and identification of those most at risk to develop the world’s most common and deadly birth defect. Completion of this project is ensured by the technical training and mentorship by the sponsorship team, the training site’s cutting-edge facilities, and education provided by the principal investigator’s M.D.-Ph.D. program.

项目摘要/摘要 近30％的胎儿酒精谱系障碍（FASD）患有先天性心脏缺陷 （CHD）。流出路（通常）缺陷过多，尽管有变化和严重性。这 大多数产前酒精暴露（PAE）研究重点是FASD，慢性饮酒和 神经系统缺陷。人口中最常见的PAE形式，是 尚未研究周期及其对心脏发育的影响。我们将研究效果 通过多机构临床研究和我们的急性Pae murine，急性，周围感受饮酒 模型，其中小鼠接受两次腹膜内注射3g/kg的30％乙醇，这一点至关重要 器官发生。我们假设PAE引起的oft缺陷的变化率和严重程度可以是 通过PAE和其他非遗传突变的结合来解释，这些突变导致一般 允许的背景。我们认为Notch途径中的突变建立了如此普遍的允许 背景，PAE通过表观遗传调节Notch与这些突变进行协同作用 通常畸形。本研究的目标1将解决PAE经常有害影响的特异性 对齐并定义了患有PAE诱导的失去缺陷的个体中破坏的分子途径。我们将 通过对PAE和CHD诊断和途径的相对风险分析以及随后的后勤状态实现这一目标 整个基因组测序数据的回归分析。 AIM 2将定义酒精驱动表观遗传调节 作为急性PAE和其他非链接凹入途径突变的机制 破坏开发。使用分子测定和组织学分析的结合，体内和体内 我们将测试这种综合的致变质损伤对第二心脏场（SHF）生存能力和能力的影响 迁移到Oft。我们将建立PAE驱动表观遗传调节，破坏Notch基因可及性 和转录，抑制Notch信号传导，并在体外使用泛酮乙酰转移酶抑制剂 证明预防PAE诱导的高乙酰化足以挽救Notch信号和SHF 生存能力。同样，通过在体外过度表达Notch的细胞内结构域（NICD）的挽救SHF生存力 巩固了急性PAE和Notch突变协同的点，因为缺口信号传导丢失了SHF生存力。 作为检查临床相关的急性PAE与心脏相互作用的首批研究之一 开发，本研究涉及NIAAA的优先级，以定义非智力暴露的​​影响 和酒精对可理解的器官系统的影响。这项研究确定的遗传途径，包括 在PAE引起的CHD中，对Notch途径的新颖验证将为疾病提供目标 预防和确定最有风险的人，发展世界上最常见和致命的先天缺陷。 赞助团队的技术培训和心态确保了该项目的完成 培训网站的尖端设施以及首席研究员的M.D.-PH.D.提供的教育程序。