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：
10115737
负责人：
SARAH J BLOSSOM
金额：
$ 27.02万
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/10115737
关键词：
Address 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 base bisulfite sequencing cytokine disorder risk drinking water environmental chemical experimental study genetic signature genome-wide in vivo inhibitor/antagonist 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.

项目概要研究描述环境污染物三氯乙烯 (TCE) 的自身免疫促进作用，我们的长期目标是关注狼疮易发小鼠的效应/记忆 CD4+ T 细胞的功能影响。是了解 TCE 如何驱动 CD4 细胞分化以促进自身免疫和/或超敏反应目标是 (i) 确定 TCE 如何通过其主要代谢物三氯乙醛。水合物 (TCAH)，改变幼稚 CD4 细胞向效应子/记忆子集的分化，对于促进（例如 Th1 和 Th17）或体外抑制自身免疫（例如 Th2 和 TREG）以及 (ii) 确定将比较这些亚型的体内致病性（MRL+/+）和自身免疫倾向（MRL+/+）。抗性（B6）小鼠品系，以了解遗传易感因素的贡献。性别可能揭示为什么女性会增加自身免疫性疾病的风险。核心假设是 TCAH。促进致病效应细胞的分化和/或减少效应细胞的扩增与抑制自身免疫有关，涉及基因表达和 DNA 甲基化的变化。尽管在 MRL+/+ 小鼠中观察到免疫病理学，但 TCAH 将产生独特的遗传特征与 B6 小鼠相比，雌性小鼠的这一结果比雄性小鼠的效果更强，其理由是现在已经很明显了。可能在 DNA 甲基化水平上调节的关键 CD4 功能效应发生在分化，因此不能在已经分化的细胞中进行研究将测试中心假设。三个具体目标：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) 确定体内分化 Th 亚群的致病性。目的，RNA-Seq 将确定 TCAH 是否直接改变可能赋予选择性的基因表达在目标 2 中，减少亚硫酸氢盐测序的代表性。用于确定分化 CD4+ T 细胞中 TCAH 诱导的 DNA 甲基化变化。使用以下方法测试 TCAH 诱导的基因表达变化是否在 DNA 甲基化水平上介导甲基供体或甲基化抑制剂会逆转重要甲基变体的存在。第三个目标将测试 Th 亚群产生免疫的能力。过继转移实验中的病理学在转移前用甲基供体处理细胞将连接。目标 1 和 2 结果确定 TCAH 是否促进 DNA 甲基化中 T 细胞介导的病理学所提出的研究具有创新性且意义重大，因为它重点关注 TCAH 如何实现。改变分化过程中产生的液体 DNA 甲基化模式，从而发现新的基因或甲基化模式可能导致 TCE 诱导的人类免疫紊乱。知识可以通过使免疫反应正常化来识别免疫介导的靶向治疗途径 TCE 暴露个体。