Identifying and protecting alcohol-sensitive epigenetic changes in congenital heart disease

识别和保护先天性心脏病中酒精敏感的表观遗传变化

• 批准号: 10301817
• 负责人: Stephanie Marie Ford
• 金额: $ 19.31万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10301817
• 关键词: 3-Dimensional; Affect; Alcohols; Betaine; Birds; Carbon; Cardiac; Cardiac development; Centers for Disease Control and Prevention (U.S.); Child; Childhood; Choline; Choline Deficiency; Clinical; Clinical Trials; Congenital Heart Defects; Craniofacial Abnormalities; Critical Pathways; DNA; DNA Methylation; DNA Sequence; Data; Defect; Development; Development Plans; Disease; Educational process of instructing; Embryo; Embryonic Heart; Epidemic; Epigenetic Process; Ethanol; Face; Family; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Fetal alcohol effects; Fluorescent in Situ Hybridization; Fostering; Future; Gene Expression; Genes; Genome; Glutathione; Growth; HAS2 gene; Head; Health; Heart; Heart Abnormalities; Heavy Drinking; High Prevalence; Human; Image; Individual; Infant; Laboratories; Lead; Life; Mentors; Mentorship; Metabolism; Methylation; Modeling; Mothers; Mus; Neurologic Effect; Newborn Infant; Optical Coherence Tomography; Outcome; Pathogenesis; Pathway interactions; Phenotype; Physicians; Pregnancy; Pregnant Women; Prevalence; Prevention; Prevention strategy; Quail; RNA; Recommendation; Research; Research Personnel; Research Training; Scientist; Site; Structure; Supplementation; Surgeon; Time; Transforming Growth Factor beta; United States National Institutes of Health; Universities; Vertebrates; Work; XCL1 gene; alcohol abstinence; alcohol consumption during pregnancy; alcohol effect; alcohol exposure; alcohol testing; animal model development; bisulfite sequencing; cardiogenesis; career; career development; choline supplementation; cohort; comparative genomics; congenital heart disorder; cost; design; dietary; experience; experimental study; gastrulation; laboratory experience; methylation pattern; mouse model; novel; optical imaging; placental mammal; prevent; septal defect; skills

This proposal describes a five-year mentored research and training plan that will facilitate the development of Dr. Stephanie Ford, MD as an independent investigator in the pathogenesis of congenital heart disease. Building upon Dr. Ford’s background as a clinical neonatologist and a basic scientist, she will attain expertise in design of mouse studies, RNA-FISH, and epigenetic mechanisms. She will gain skills through structured mentorship, hands-on laboratory experiences, didactic teaching, and formal classwork at Case Western Reserve University, FAES at the NIH, and Jackson Laboratories. Dr. Michael Jenkins, a pioneer in cardiac optical imaging, and Dr. Cynthia Bearer, an expert in prenatal alcohol exposure models, will provide their expertise and mentorship skills to this project, fostering Dr. Ford’s transition to research independence. An estimated 2.4-4.8% of newborns in the U.S. have fetal alcohol spectrum disorders (FASDs), caused by prenatal alcohol exposure (PAE). PAE induced Congenital Heart Diseases (CHDs) have not been studied as intensively as other FASD outcomes despite their high prevalence rate (40%). The CHDs associated with FASDs, mostly valvuloseptal and outflow tract defects, are life-threatening and impact growth and health. PAE is known to affect methylation and one-carbon metabolism. Normal one-carbon metabolism and its resulting methylation of DNA is crucial for the correct expression of genes. Comparative genomics studies have revealed that there is strong epigenetic conservation across vertebrate species including mice and avians, particularly the hyper-and hypo-methylated DNA sequences of critical genes. We will investigate the PAE-induced changes at times critical to heart development (endocardial cushion and 4 chamber development) in mouse and avian embryonic hearts. All hearts will be imaged with optical coherence tomography to rapidly determine their phenotype. DNA methylation changes will be determined with a combination of methyl-ATAC-seqand bisulfite sequencing. DNA methylation will be compared in both species, as conserved changes in two species are more likely to be relevant to human PAE-induced defects. RNA-FISH will be used to confirm gene expression changes, which will allow us to pinpoint where within the 3D heart, such as the forming valves, gene expression is changing. We will then explore the use of choline and glutathione to prevent the effects of PAE. Choline and glutathione are known to promote methylation in one-carbon metabolism. Choline has been shown in human studies to prevent early neurologic effects of PAE. We have shown in an avian model that glutathione prevents the CHDs and abnormal DNA methylation seen after PAE. We will use both avian and mouse models to determine the effects of alcohol + choline or glutathione on cardiac structure, DNA methylation, and gene expression. We hypothesize that by maintaining normal methylation, and therefore DNA expression, our chosen compounds will prevent the CHDs that result from PAE. Compounds that could prevent PAE-induced CHDs could help thousands of children and their families each year.

该提案描述了一项五年期指导研究和培训计划， Stephanie福特博士作为先天性心脏病发病机制的独立研究者的发展 疾病凭借福特博士作为临床病理学家和基础科学家的背景，她将获得 在小鼠研究设计、RNA-FISH和表观遗传机制方面的专业知识。她将通过以下方式获得技能： 结构化的指导，动手实验室经验，教学式教学，并在凯斯正式课堂作业 西储大学，美国国立卫生研究院的FAES和杰克逊实验室。迈克尔·詹金斯博士， 心脏光学成像和辛西娅·贝尔博士，产前酒精暴露模型专家，将提供 他们的专业知识和指导技能，这个项目，促进博士福特的过渡到研究的独立性。 据估计，美国2.4-4.8%的新生儿患有胎儿酒精谱系障碍（FASD），由以下原因引起： 产前酒精暴露（PAE）。尚未研究PAE诱导的先天性心脏病（CHD）， 尽管FASD的患病率很高（40%），但其与其他FASD结局一样强烈。相关的CHD FASD，主要是瓣膜间隔和流出道缺陷，危及生命，影响生长和健康。PAE 影响甲基化和一碳代谢。正常的一碳代谢及其产生的 DNA的甲基化对于基因的正确表达至关重要。比较基因组学研究显示 在包括小鼠和鸟类在内的脊椎动物物种中， 关键基因的高和低甲基化DNA序列。我们将研究PAE诱导的变化， 小鼠和禽类心脏发育的关键时间（内膜垫和4室发育） 胚胎心脏所有的心脏都将通过光学相干断层扫描成像，以快速确定其 表型DNA甲基化变化将用甲基-ATAC-seq和亚硫酸氢盐的组合测定。 测序DNA甲基化将在两个物种中进行比较，因为两个物种中的保守变化更多。 可能与人类PAE诱导的缺陷相关。RNA-FISH将用于确认基因表达变化， 这将使我们能够精确定位3D心脏中的哪些位置，例如正在形成的瓣膜，基因表达正在发生变化。 然后，我们将探索使用胆碱和谷胱甘肽来预防PAE的影响。胆碱和 已知谷胱甘肽促进一碳代谢中的甲基化。胆碱已被证明在人类 预防PAE的早期神经学影响的研究。我们在鸟类模型中表明，谷胱甘肽可以预防 PAE后出现CHD和异常DNA甲基化。我们将使用鸟类和小鼠模型， 确定酒精+胆碱或谷胱甘肽对心脏结构、DNA甲基化和基因的影响。 表情我们假设通过维持正常的甲基化， 表达，我们选择的化合物将防止由PAE引起的CHD。的化合物 每年可以帮助成千上万的儿童和他们的家庭。