Maternal genotype, choline intervention,& epigenetics in Fetal Alcohol Syndrome

母体基因型、胆碱干预、

• 批准号: 9032100
• 负责人: Daniel Goldowitz
• 金额: $ 30.79万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-10 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9032100
• 关键词: Address; Affect; Alcohol or Other Drugs use; Alcohol-Induced Neurotoxicity; Bioinformatics; Brain; Brain Injuries; Brain Stem; CASP3 gene; Cell Death; Child; Choline; Collaborations; Data; Development; Dose; Early Intervention; Embryo; Embryo Transfer; Environment; Environmental Impact; Epigenetic Process; Ethanol; Evaluation; Female; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Fetal Alcohol Syndrome; Fetus; Functional disorder; Gene Expression; Generations; Genes; Genetic; Genome; Genotype; Heterozygote; Hour; Human; In Situ Nick-End Labeling; Inbred Strains Mice; Individual; Intervention; Label; Laboratories; Life; Mediating; Mediator of activation protein; Metabolic Pathway; Metabolism; Methods; Modification; Molecular; Molecular Analysis; Mothers; Mus; Nervous system structure; Neural tube; Neuraxis; Operative Surgical Procedures; Outcome; Play; Population; Positioning Attribute; Predisposition; Process; Protocols documentation; Recombinant Inbred Strain; Research; Resources; Risk; Role; Series; Severities; Techniques; Telencephalon; Testing; Therapeutic; Transplantation; Treatment Efficacy; Validation; Work; alcohol effect; alcohol exposure; alcohol sensitivity; base; bisulfite; bisulfite sequencing; choline supplementation; design; epigenome; fetal; genome wide methylation; high risk; imprint; mRNA Expression; male; member; neuron loss; neurotoxicity; public health relevance; pyrosequencing; research study; response; treatment strategy

 DESCRIPTION (provided by applicant): The type and severity of ethanol-induced alterations following prenatal ethanol exposure is strongly impacted by genetics. However, the role of genetics is complicated by the fact that both the mother and fetus have unique genotypes. The role of genetics could be an even bigger consideration in the evaluation of treatments, since the metabolism of any therapeutic is completely regulated by the mother in early development. This proposal will test the following hypotheses: 1) the genotype of the mother has a significant role in determining the level of ethanol-induced cell death in the developing brain as well as in the efficacy of choline treatment in modulating ethanol-induced cell death and 2) the epigenome of the embryo has an important role in modulating genetic differences in susceptibility to the effects of ethanol exposure. We have been examining the BXD panel of mice, generated by crossing C57BL/6J (B6) and DBA/2J (D2) strains, and show differential sensitivity following equivalent ethanol exposure thereby facilitating the testing of these hypotheses. Specific Aim 1 will test the hypothesis that the maternal genotype influences the level of ethanol-induced cell death in the developing neural tube. Two complementary approaches, reciprocal crosses and embryo transplants, will be used. Embryos will be exposed to ethanol on embryonic day 9 and collected 7 hours after the initial ethanol exposure. Cell death will be quantified from TUNEL and activated caspase-3 labeled sections in the developing telencephalon and brain stem. If, in both approaches, the level of cell death is altered depending upon the genotype of the mother, it will show that the maternal genotype is an important mediator. Specific Aim 2 will test the hypothesis that there are genetic differences in the dam that contribute to the efficacy of choline in mitigating ethanol's effects on cell death. B6 and BXD strains that show high levels of cell death following ethanol exposure will be tested with different doses of choline to find the lowest dose that is efficacious in ameliorating ethanol's effects. If the dose differs significantly acros the strains it will demonstrate that genotype is critical. In contrast, if choline's effects are equivalent across strains, it will suggest that genotype is unimportant and that choline should be equally beneficial in all FASD children and that once a relevant dose is found within the human population, it will likely be equally effective across a wide sector of the population. Specific Ai 3 will test the hypotheses that epigenetic changes induced by ethanol exposure differ based on genotype, as well as maternal environment, thus contributing to genetic variability in susceptibility to ethanol- induced neurotoxicity. Global methylation will be examined in the telencephalon using reduced representation bisulfite sequencing and bisulfite pyrosequencing and be compared to changes in gene expression. Information from this proposal will be important for determining 1) which genome to examine to identify children most at risk for specific types of ethanol-induced neuroteratological effects and 2) whether genotype needs to be considered in implementing choline as a therapeutic for ethanol-induced brain damage.

 描述（由适用提供）：遗传学对乙醇诱导的改变后乙醇诱导的改变的类型和严重程度受到遗传学的强烈影响。但是，母亲和胎儿都有独特的基因型，遗传学的作用变得复杂。遗传学的作用在评估治疗中可能是更大的考虑因素，因为任何治疗性的代谢在早期发育中都完全受到母亲的调节。该提议将检验以下假设：1）母亲的基因型在确定发育中的大脑中乙醇诱导的细胞死亡水平以及胆碱治疗在调节乙醇诱导的细胞死亡中的有效性和2）胚胎的表观组在调节乙醇中具有乙醇差异的作用在乙醇中具有重要作用。我们一直在检查通过C57BL/6J（B6）和DBA/2J（D2）菌株产生的BXD小鼠面板，并在等效乙醇暴露后显示出差异敏感性，从而支持这些假设的测试。具体目标1将检验以下假设：乙醇诱导的神经管中细胞死亡的水平。将使用两种完整的方法，即相互交叉和胚胎移植。胚胎将在胚胎第9天暴露于乙醇，并在初始乙醇暴露后7小时收集。细胞死亡将从TUNEL和激活的caspase-3在发育中的型脑词干和脑干中进行量化。如果在这两种方法中，细胞死亡的水平都根据母亲的基因型而改变，则将表明母体基因型是重要的介体。具体目标2将检验以下假设：大坝中存在遗传差异，这有助于胆碱的效率 减轻乙醇对细胞死亡的影响。乙醇暴露后显示高水平细胞死亡的B6和BXD菌株将用不同剂量的胆碱进行测试，以找到有效地改善乙醇作用的最低剂量。如果在各种菌株之间的剂量不同，它将证明基因型至关重要。相比之下，如果胆碱的作用在各种菌株之间等效，则表明基因型是不重要的，并且胆碱在所有FASD儿童中都应同样有益，并且一旦在人口中发现相关剂量，它可能同样有效，在广泛的人群中。特定的AI 3将检验假设，即乙醇暴露引起的表观遗传变化基于基因型和MATER环境不同，从而导致对乙醇诱导的神经毒性的易感性的遗传变异性。将使用降低的代表性亚硫酸盐测序和亚硫酸硫酸硫酸硫酸硫酸硫酸盐磷酸测序，并将其与基因表达的变化进行比较。该提案中的信息对于确定1）要检查哪种基因组以确定对特定类型的乙醇诱导的神经疾病的风险最大的儿童； 2）在实施胆碱作为乙醇诱导的脑损伤的治疗方面是否需要考虑基因型。