Role of SRY in transgenerational transmission of alcohol epigenetic marks on proopiomelanocortin gene

SRY在阿黑皮素原基因酒精表观遗传标记跨代传递中的作用

• 批准号: 9382377
• 负责人: DIPAK KUMAR SARKAR
• 金额: $ 34.88万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382377
• 关键词: Address; Affect; Alcohol consumption; Alcohol-Related Disorders; Alcohols; Animal Model; Animals; Behavioral; Binding; Binding Sites; Biological Markers; Child; Complex; Corticosterone; DNA; Data; Development; Disease; Drug abuse; Drug usage; Electroporation; Emotional Disturbance; Endocrine; Environmental Risk Factor; Epigenetic Process; Ethanol; Excision; Fertilization; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Fetal Alcohol Syndrome; Fetal alcohol effects; Gametogenesis; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genomic DNA; Germ Cells; Grant; Growth; Health; Heritability; Hormones; Human; Hypothalamic structure; Immune; Immune System Diseases; Immune System and Related Disorders; Immune response; Inbred F344 Rats; Interferon Type II; Interferons; Knowledge; Learning Disabilities; Life; Link; Malignant Neoplasms; Mediating; Medical; Methods; Methylation; Modeling; Modification; Molecular; Mothers; Mutation Analysis; National Institute on Alcohol Abuse and Alcoholism; Neuropeptides; Neurosecretory Systems; POMC gene; Parents; Patients; Pharmaceutical Preparations; Phenotype; Plasmids; Population; Pro-Opiomelanocortin; Process; Production; Promoter Regions; Public Health; Rattus; Reporter; Research; Risk; Rodent Model; Role; Series; Stress; Techniques; Testing; Third Pregnancy Trimester; Tissues; Transfection; Variant; Y Chromosome; alcohol and other drug; alcohol effect; alcohol exposure; base; beta-Endorphin; biological adaptation to stress; chromatin immunoprecipitation; cytokine; demethylation; drug induced behavior; endophenotype; epidemiology study; expression vector; fetal; grandparent; immune function; immune system function; imprint; in vivo; interest; knock-down; male; melanocortin receptor; methylation pattern; next generation; novel; offspring; overexpression; peptide B; preference; prenatal; prevent; promoter; pyrosequencing; response; sex; sperm cell; sry Genes; transcription factor; transgenerational epigenetic inheritance; transmission process; young adult

Project Summary/Abstract Children and young adults who were exposed to alcohol during fetal life often show fetal alcohol spectrum disorders (FASD) consisting of learning disabilities, emotional disturbances, stress abnormalities and immune incompetence. Furthermore, some grandparents who had alcohol-related medical problems often had grandchildren with a higher rate of fetal alcohol syndrome than those grandparents without alcohol-related medical problems. Epidemiological studies also have shown association of paternal alcohol consumption with deficiencies in offspring—effects which are typically found with maternal alcohol exposure. However, the process underlying the heritability of alcohol-induced health issues is not clearly understood. Our recent studies identified an epigenetic mechanism in the heritable effect of fetal alcohol on stress response abnormalities and immune dysfunctions. These studies showed that the proopiomelanocortin (Pomc) gene, which negatively controls the functions of the stress axis and stimulates the functions of the immune systems, is hypermethylated while the response of stress axis hormones and the production of the cytokine interferon-γ (IFN-γ) is dysregulated in fetal alcohol exposed offspring for three generations via the male germline. However, the process by which these epigenetic variants induced by fetal alcohol are transmitted from parents to offspring through multiple generations has not been established. We hypothesize that a transcription factor like sex-determining region Y (SRY), which has significant control over Pomc gene expression, undergoes heritable epigenetic modifications by fetal alcohol exposures that in turn modify the expression of the Pomc gene and its endophenotypes. The objective of this proposal is to determine the heritable epigenetic effects of alcohol exposure during the prenatal period on SRY-mediated Pomc gene expression and its endophenotypes, i.e., the stress hormone response and IFN-γ production to an immune challenge, in isogenic Fischer 344 rats. This research aim will be achieved by determining the relationship between the fetal alcohol epigenetic marks on Pomc and its endophenotypes with the epigenetic marks on the Sry gene in the isogenic rat population in three generations of offspring, studying the interaction between SRY and Pomc at the molecular level, and evaluating whether increased methylation affects SRY interaction with the Pomc promoter and contributes to fetal alcohol effects on Pomc and its endophenotypes. Various state of the art techniques will be employed involving promoter methylation, promoter demethylation, promoter activity, mutational analysis, gene knock down, in vivo electroporation for gene overexpression, and stress and immune response studies. These studies will establish the process by which developmental alcohol exposure induces gene expression variants in the offspring and how they are transmitted to next generations. Knowledge on the process involved in the heritability of alcohol-induced epigenetic variants may help in establishing new biomarkers and novel epigenetic-based therapies for many alcohol-related disorders.

项目总结/摘要 在胎儿期暴露于酒精的儿童和年轻人经常显示出胎儿酒精谱 FASD包括学习障碍、情绪障碍、应激异常和免疫功能障碍。 无能此外，一些有与酒精有关的医疗问题的祖父母经常 与没有酒精相关的祖父母相比，有更高的胎儿酒精综合征发生率的孙辈 医疗问题。流行病学研究还表明，父亲饮酒与 后代的缺陷-母亲接触酒精通常会产生这种影响。但 酒精引起的健康问题的遗传性的潜在过程还不清楚。我们最近 研究确定了胎儿酒精对应激反应的遗传效应的表观遗传机制 异常和免疫功能障碍。这些研究表明，阿黑皮素原（Pomc）基因， 其消极地控制应激轴的功能并刺激免疫系统的功能， 当应激轴激素的反应和细胞因子干扰素-γ的产生 (IFN-γ）在胎儿酒精暴露后代中通过雄性生殖系失调三代。然而，在这方面， 这些由胎儿酒精诱导的表观遗传变异从父母传递到 多代的后代尚未建立。我们假设一个转录因子， 性别决定区Y（SRY）对Pomc基因的表达具有重要的控制作用， 胎儿酒精暴露引起的可遗传表观遗传修饰，反过来又改变了Pomc基因的表达， 基因及其内表型。这项建议的目的是确定遗传的表观遗传效应， 产前酒精暴露对SRY介导的Pomc基因表达及其内在表型的影响， 也就是说，等基因Fischer 344大鼠对免疫挑战的应激激素反应和IFN-γ产生。 本研究的目的将通过确定胎儿酒精表观遗传标记之间的关系来实现 用Sry基因上的表观遗传标记对Pomc及其内表型的影响 三代后代，在分子水平上研究SRY和Pomc之间的相互作用， 评估增加的甲基化是否影响SRY与Pomc启动子的相互作用， 胎儿酒精对Pomc及其内表型的影响将采用各种现有技术 涉及启动子甲基化、启动子去甲基化、启动子活性、突变分析、基因敲除 向下，用于基因过表达的体内电穿孔，以及应激和免疫应答研究。这些 研究将确定发育期酒精暴露诱导基因表达变异的过程 以及它们如何传递给下一代。对参与的过程的了解 酒精诱导的表观遗传变异的遗传性可能有助于建立新的生物标志物和新的 基于表观遗传学的疗法治疗许多酒精相关疾病。