Epigenetic modulation of CD4+ T cell differentiation and autoimmunity by trichloroethylene

三氯乙烯对 CD4 T 细胞分化和自身免疫的表观遗传调节

基本信息

批准号：
10392944
负责人：
SARAH J BLOSSOM
金额：
$ 26.86万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10392944
关键词：
Adoptive Transfer Autoimmune Autoimmune Diseases Autoimmunity CD4 Positive T Lymphocytes Cell Differentiation process Cells Chemicals Chronic Control Groups DNA Methylation Dermal Disease Effector Cell Environmental Pollutants Epigenetic Process Event Exposure to Female Flow Cytometry Gene Expression Genes Genetic Predisposition to Disease Goals Grant Half-Life Human Hydration status Hypersensitivity Immune Immune System Diseases Immune response In Vitro Individual Industrialization Inflammatory Interferons Knowledge Liquid substance Lupus Mediating Memory Metabolism Methylation Mouse Strains Mus Nature Outcome Pathogenicity Pathology Pathway interactions Prevention Process Production Promoter Regions Proteins Public Health Research Resistance Risk Factors Solvents T cell differentiation T-Cell Activation T-Lymphocyte Testing Time Tissues Trichloroethylene Variant bisulfite sequencing cytokine disorder risk drinking water environmental chemical experimental study genetic signature genome-wide in vivo inhibitor innovation insight lupus cutaneous lupus prone mice lupus-like male memory CD4 T lymphocyte methylation pattern novel sex superfund site targeted treatment transcriptome sequencing trichloroacetaldehyde water solubility

项目摘要

PROJECT SUMMARY Studies to delineate the autoimmune-promoting effects of the environmental pollutant, trichloroethylene (TCE), have focused on functional effects in effector/memory CD4+ T cells from lupus-prone mice. Our long-term goal is to understand how TCE drives CD4 cell differentiation to promote autoimmunity and/or hypersensitivity disorders. The objectives are (i) to determine how TCE through its primary metabolite, trichloroacetaldehyde hydrate (TCAH), alters differentiation of naïve CD4 cells to effector/memory subsets important in promoting (e.g., Th1 and Th17) or suppressing autoimmunity (e.g., Th2 and TREG) in vitro and (ii) to determine pathogenicity of these subsets in vivo. CD4s will be compared in both autoimmune-prone (MRL+/+) and resistant (B6) strains of mice to understand the contribution of genetic susceptibility factors. Inclusion of both sexes may reveal why being female elevates autoimmune disease risk. The central hypothesis is that TCAH promotes the differentiation of pathogenic effector cells and/or decreases expansion of effector cells associated with suppression of autoimmunity involving changes in gene expression and DNA methylation. TCAH will generate unique genetic signatures although immune pathology will be observed in MRL+/+ mice rather than in B6 mice, that will be more robust in females vs. males. The rationale is that it is now apparent that key CD4 functional effects that may be regulated at the level of DNA methylation occur during differentiation, and thus cannot be studied in already differentiated cells. The central hypothesis will be tested in three specific aims: 1) Define effects of TCAH on CD4+ T cell differentiation by assessing gene expression and 2) DNA methylation and 3) Determine pathogenicity of differentiating Th subsets in vivo. Under the first Aim, RNA-Seq will determine whether TCAH directly alters the expression of genes that may confer a selective advantage to pathogenic effector CD4+ T cells. In Aim 2, reduced representation bisulfite sequencing will be used to determine TCAH-induced changes in DNA methylation in differentiating CD4+ T cells. Experiments will test whether gene expression changes induced by TCAH are mediated at the level of DNA methylation using either methyl donors or methylation inhibitors to reverse effects. The presence of important methyl variants will be determined by flow cytometry. The third Aim will test Th subsets for their ability to generate immune pathology in adoptive transfer experiments. Treatment of cells with methyl donors prior to transfer will connect Aim 1 and 2 results to determine whether TCAH promotes T cell-mediated pathology in a DNA methylation- dependent manner. The proposed research is innovative and significant because it focuses on how TCAH alters the fluid DNA methylation pattern generated during differentiation that will lead to the discovery of novel gene or methylation patterns that may be responsible for TCE-induced immune disorders in humans. Such knowledge may identify immune-mediated pathways for targeted therapy by normalizing immune responses in TCE-exposed individuals.

项目摘要描述环境污染物三氯乙烯（TCE）的自身免疫性促进作用的研究已重点关注效应器/记忆CD4+ T细胞的功能效应。我们的长期目标是要了解TCE如何驱动CD4细胞分化以促进自身免疫和/或超敏反应疾病。目的是（i）确定TCE如何通过其主要代谢物三氯乙醛水合（TCAH），改变CD4细胞对效应子/记忆子集的分化对促进（例如，Th1和Th17）或在体外抑制自身免疫（例如Th2和Treg），并且（ii）确定这些子集在体内的致病性。将在自身免疫性（MRL+/+）和抗性（B6）小鼠菌株了解遗传敏感性因子的贡献。两者都包含性别可能会揭示为什么女性会增加自身免疫性疾病风险。中心假设是TCAH 促进致病效应细胞的分化和/或降低效应细胞的膨胀与抑制自身免疫性有关，涉及基因表达和DNA甲基化的变化。尽管将在MRL+/+小鼠中观察到免疫病理学，但TCAH将产生独特的遗传特征而不是在B6小鼠中，这将在女性与男性中更强大。理由是现在很明显在DNA甲基化水平上可能调节的关键CD4功能效应发生在期间分化，因此无法在已经分化的细胞中进行研究。中央假设将进行测试在三个具体目的中：1）通过评估基因表达来定义TCAH对CD4+ T细胞分化的影响 2）DNA甲基化和3）确定在体内分化亚群的致病性。在第一个目的，RNA-Seq将确定TCAH是否直接改变了可能会有选择性的基因的表达致病效应子CD4+ T细胞的优势。在AIM 2中，降低表示Bisulfite测序将是用于确定TCAH诱导的分化CD4+ T细胞中DNA甲基化的变化。实验会测试使用TCAH诱导的基因表达是否在DNA甲基化水平上介导甲基供体或甲基化抑制剂要反向作用。重要的甲基变体的存在将通过流式细胞仪确定。第三个目标将测试TH子集的产生免疫力的能力自适应转移实验中的病理。转移前用甲基供体处理细胞将连接 AIM 1和2的结果是确定TCAH是否促进了DNA甲基化中T细胞介导的病理依赖方式。拟议的研究具有创新性和重要意义，因为它重点是TCAH 改变在分化过程中产生的流体DNA甲基化模式，这将导致发现新颖可能导致人类TCE诱导的免疫疾病的基因或甲基化模式。这样的知识可以通过标准化免疫反应来确定针对靶向治疗的免疫介导的途径暴露于TCE的个体。