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.

项目概要/摘要 近 30% 的胎儿酒精谱系障碍 (FASD) 患者患有先天性心脏缺陷 (CHD) 流出道 (OFT) 缺陷的比例较高，但其发生率和严重程度各不相同。 大多数产前酒精暴露 (PAE) 研究重点关注胎儿酒精谱系障碍 (FASD)、长期饮酒和 神经系统缺陷是人群中最常见的 PAE 形式，是在急性暴露期间发生的。 围孕期及其对心脏发育的影响尚未研究。我们将研究其影响。 通过多机构临床研究和我们的急性 PAE 小鼠对急性围孕期饮酒的影响 模型中，小鼠在心脏功能关键点接受两次腹腔注射 3g/kg 30% 乙醇 我们探讨了 PAE 引起的 OFT 缺陷的不同发生率和严重程度。 PAE 和其他非有害突变的组合可以解释这一点，这些突变会导致基因突变 我们相信Notch通路的突变建立了这样的遗传许可。 PAE 通过 Notch 的表观遗传调控与这些突变协同作用，从而产生 本研究的目标 1 将解决 PAE 对 OFT 有害影响的特异性。 我们将对齐并定义 PAE 引起的 OFT 缺陷个体中被破坏的分子途径。 通过 PAE 和 CHD 诊断和途径的相对风险分析以及后续的后勤保障来实现这一目标 目标 2 将定义酒精驱动的表观遗传调控 作为急性 PAE 和其他非有害 Notch 途径突变协同作用的机制 结合体外和体内分子测定和组织学分析，破坏 OFT 发育。 我们将测试这种联合致畸损伤对第二心脏区 (SHF) 活力和能力的影响 我们将建立 PAE 驱动的表观遗传调控，破坏 Notch 基因的可及性。 和转录，抑制 Notch 信号传导，并在体外使用泛组蛋白乙酰转移酶抑制剂 证明预防 PAE 诱导的过度乙酰化足以挽救 Notch 信号传导和 SHF 同样，通过在体外过度表达缺口胞内结构域（NICD）来挽救 SHF 的活力。 巩固急性 PAE 和 Notch 突变协同作用的观点，因为 Notch 信号传导的丧失会抑制 SHF 的生存能力。 作为首批研究临床相关急性 PAE 与心脏相互作用的研究之一 发展，本研究解决了 NIAAA 的优先事项，以确定非慢性暴露的影响 和”酒精对尚未研究的器官系统的影响，包括本研究确定的遗传途径。 Notch 通路在 PAE 诱发的冠心病中具有重要意义的新验证将为疾病提供靶标 预防和识别那些最有可能患上世界上最常见和致命的出生缺陷的人。 赞助团队的技术培训和指导确保了该项目的完成 培训中心的尖端设施以及首席研究员的医学博士项目提供的教育。