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（由申请人提供）：产前乙醇暴露后乙醇引起的改变的类型和严重程度受到遗传的强烈影响。然而，由于母亲和胎儿都具有独特的基因型，遗传学的作用变得复杂。在评估治疗方法时，遗传学的作用可能是一个更大的考虑因素，因为任何治疗方法的新陈代谢在早期发育过程中都完全由母亲调节。该提案将测试以下假设：1）母亲的基因型在确定发育中大脑中乙醇诱导的细胞死亡水平以及胆碱治疗调节乙醇诱导的细胞死亡的功效方面具有重要作用；2）胚胎的表观基因组在调节对乙醇暴露影响的易感性的遗传差异方面具有重要作用。我们一直在检查通过 C57BL/6J (B6) 和 DBA/2J (D2) 品系杂交产生的 BXD 小鼠组，并在等量乙醇暴露后显示出不同的敏感性，从而促进了这些假设的测试。具体目标 1 将检验母体基因型影响发育中神经管中乙醇诱导的细胞死亡水平的假设。将使用两种互补的方法，即相互杂交和胚胎移植。胚胎将在胚胎第 9 天暴露于乙醇，并在初次乙醇暴露后 7 小时收集。细胞死亡将从发育中的端脑和脑干中的 TUNEL 和激活的 caspase-3 标记切片进行量化。如果在这两种方法中，细胞死亡水平根据母亲的基因型而改变，则表明母亲的基因型是一个重要的介质。具体目标 2 将检验以下假设：母鼠中存在有助于胆碱功效的遗传差异 减轻乙醇对细胞死亡的影响。将使用不同剂量的胆碱对暴露于乙醇后表现出高水平细胞死亡的 B6 和 BXD 菌株进行测试，以找到有效改善乙醇影响的最低剂量。如果不同菌株的剂量显着不同，则表明基因型至关重要。相比之下，如果胆碱在不同菌株中的作用相同，则表明基因型并不重要，胆碱对所有 FASD 儿童应该同样有益，而且一旦在人群中发现相关剂量，它可能在广泛的人群中同样有效。具体的 Ai 3 将测试以下假设：乙醇暴露引起的表观遗传变化因基因型和母体环境而异，从而导致对乙醇引起的神经毒性的易感性的遗传变异。将使用简化代表性亚硫酸氢盐测序和亚硫酸氢盐焦磷酸测序来检查端脑中的整体甲基化，并与基因表达的变化进行比较。该提案中的信息对于确定以下方面非常重要：1）要检查哪个基因组，以识别最有可能遭受特定类型的乙醇引起的神经畸形影响的儿童；2）在使用胆碱作为乙醇引起的脑损伤的治疗剂时是否需要考虑基因型。