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

• 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
• 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

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）的自身免疫促进作用的研究， 已经集中于来自狼疮易感小鼠的效应/记忆CD 4 + T细胞的功能效应。我们的长期目标 是了解TCE如何驱动CD 4细胞分化以促进自身免疫和/或超敏反应 紊乱目的是（i）确定三氯乙烯如何通过其主要代谢产物三氯乙醛 水合物（TCAH），改变幼稚CD 4细胞分化为效应/记忆亚群，这在促进 (e.g., Th 1和Th 17）或抑制自身免疫（例如，Th 2和TREG）和（ii）测定 这些亚群在体内的致病性。将在自身免疫易感（MRL+/+）和 耐药（B6）小鼠品系，以了解遗传易感性因素的贡献。既包括 性别可能揭示了为什么女性会增加自身免疫性疾病的风险。中心假设是TCAH 促进致病性效应细胞的分化和/或减少效应细胞的扩增 与涉及基因表达和DNA甲基化变化的自身免疫抑制相关。 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 而不是在B6小鼠中，这将在雌性小鼠中比雄性小鼠更强大。理由是现在很明显 可能在DNA甲基化水平上调节的关键CD 4功能效应发生在 在分化过程中，细胞不能分化，因此不能在已经分化的细胞中进行研究。中心假设将被检验 在三个具体目标：1）通过评估基因表达来确定TCAH对CD 4 + T细胞分化的影响 （2）DNA甲基化;（3）体内检测分化型Th亚群的致病性。根据第一项 目的是，RNA-Seq将确定TCAH是否直接改变基因的表达，这些基因可能赋予选择性的免疫应答。 对致病性效应CD 4 + T细胞有利。在目标2中，减少代表性亚硫酸氢盐测序将是 用于测定TCAH诱导的分化中CD 4 + T细胞DNA甲基化的变化。实验将 测试TCAH诱导的基因表达变化是否在DNA甲基化水平介导， 甲基供体或甲基化抑制剂来逆转效应。重要的甲基变体的存在将 通过流式细胞术测定。第三个目标将测试Th亚群产生免疫应答的能力。 病理学在过继转移实验中的应用在转移之前用甲基供体处理细胞将连接 目的1和2的结果是确定TCAH是否在DNA甲基化中促进T细胞介导的病理学。 依赖的方式。该研究具有创新性和重要意义，因为它侧重于TCAH如何 改变了分化过程中产生的流体DNA甲基化模式，这将导致发现新的 基因或甲基化模式可能导致TCE诱导的人类免疫疾病。等 知识可以通过使免疫应答正常化来识别用于靶向治疗的免疫介导途径， 三氯乙烯接触者。