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

母体基因型、胆碱干预、

基本信息

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
项目类别：
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

项目摘要

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，由于母亲和胎儿都有独特的基因型，遗传的作用变得复杂。在评估治疗方法时，遗传学可能是一个更大的考虑因素，因为任何治疗方法的代谢在早期发育中都完全由母亲调节。该提议将检验以下假设：1）母亲的基因型在决定中起着重要作用。发育中大脑中乙醇诱导的细胞死亡的水平以及胆碱治疗在调节乙醇诱导的细胞死亡方面的功效；2) 胚胎的表观基因组在调节对酒精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，并在同等乙醇暴露后显示出不同的敏感性，从而促进了乙醇暴露的测试。这些假设。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。胚胎第 9 天和初次乙醇暴露后 7 小时收集的细胞死亡将从发育中的端脑和脑干中的 TUNEL 和激活的 caspase-3 标记切片进行定量。细胞死亡水平根据母亲的基因型而改变，这将表明母亲的基因型是一个重要的介质，具体目标 2 将检验母体中存在有助于胆碱功效的遗传差异的假设。在乙醇暴露后表现出高水平细胞死亡的 B6 和 BXD 菌株将用不同剂量的胆碱进行测试，以找到有效改善乙醇影响的最低剂量（如果剂量之间存在显着差异）。相反，如果胆碱的作用在不同菌株之间是相同的，则表明基因型并不重要，胆碱对所有菌株都应该同样有益。 FASD 儿童，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，特定 Ai 3 将检验乙醇暴露引起的表观遗传变化因基因型而异的假设。母体环境，从而导致对乙醇诱导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在端脑中检查全局甲基化，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效应的儿童；2）在使用胆碱作为乙醇引起的脑部治疗时是否需要考虑基因型损害。