The Role of Myeloid Cell-Specific Ten-Eleven Translocase 2 (TET2) in Regulation of Neuroinflammation

骨髓细胞特异性 10-11 转位酶 2 (TET2) 在神经炎症调节中的作用

• 批准号: 10378505
• 负责人: Ashley Munie Gardner
• 金额: $ 3.23万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378505
• 关键词: Address; Adoptive Transfer; Aggressive Clinical Course; Animal Model; Anti-Inflammatory Agents; Antigen Presentation; Autoimmune; Blocking Antibodies; Blood; Bone Marrow; CD4 Positive T Lymphocytes; CD86 gene; CNS autoimmune disease; CNS autoimmunity; Cell physiology; Cells; Cellular Infiltration; ChIP-seq; Clinical; Cytokine Activation; Cytosine; DNA; Data; Demyelinations; Development; Dioxygenases; Disease; Down-Regulation; Enterobacteria phage P1 Cre recombinase; Enzymes; Epigenetic Process; Etiology; Experimental Autoimmune Encephalomyelitis; Family; Flow Cytometry; Fluorescence-Activated Cell Sorting; Frequencies; Gene Expression; Genetic; Genetic Predisposition to Disease; Genetic Transcription; Goals; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Interleukin-1 beta; Interleukin-4; Interleukin-6; Knock-out; Laboratories; Lead; Malignant Neoplasms; Measures; Mediating; Methylation; Microarray Analysis; Modification; Multiple Sclerosis; Multiple Sclerosis Lesions; Mus; Myelin; Myelogenous; Myeloid Cell Activation; Myeloid Cells; Nervous System Trauma; Neuraxis; Nucleic Acid Regulatory Sequences; Outcome; PTPRC gene; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phagocytosis; Phenotype; Play; Population; Production; Property; Regulation; Reporter; Research; Role; Severities; Spleen; Susceptibility Gene; System; Tamoxifen; Time; Transcript; Transgenic Mice; alpha ketoglutarate; central nervous system demyelinating disorder; chemokine; conditional knockout; cytokine; demethylation; disability; experience; genetic risk factor; interest; loss of function mutation; member; mouse model; multiple sclerosis patient; multiple sclerosis treatment; neuroimmunology; neuroinflammation; neuropathology; neurotoxicity; new therapeutic target; novel; oxidation; programmed cell death ligand 1; promoter; single-cell RNA sequencing; therapeutic target; translocase

PROJECT SUMMARY Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS) believed to be autoimmune in etiology. Analysis of MS lesions, as well as studies in experimental autoimmune encephalomyelitis (EAE), a murine model of MS, have indicated that myeloid cells are likely a critical component of the pathogenesis of CNS autoimmunity. Recently, Ten-Eleven Translocase 2 (TET2) was identified as a genetic susceptibility locus for MS. TET2 is a Fe(III)-, α-ketoglutarate-dependent enzyme that catalyzes the oxidation of methylated cytosine to 5’-hydroxymethyl cytosine (5hmC) and promotes active demethylation. 5hmC is a stable epigenetic mark that can either activate or repress gene expression in a cell- and loci-dependent manner. There is growing evidence that TET2 acts as a negative regulator of myeloid cell activation and function, promoting a homeostatic or anti-inflammatory cell state. My hypothesis is that TET2 restrains the pro- inflammatory functions of myeloid cells to limit inflammation, neurological damage, and disability during CNS autoimmunity. Supporting this hypothesis, TET2 and 5hmC were found to be reduced in peripheral blood mononuclear cells of MS patients when compared with controls. Preliminary data from our laboratory demonstrates that the reduction of TET2 and 5hmC in peripheral blood mononuclear cells is recapitulated in EAE, and that TET2 transcripts and 5hmC content are further reduced in CNS-infiltrating myeloid cells compared with the periphery. Additionally, adoptive transfer of encephalitogenic Th17 cells into Tet2+/- mice led to an exacerbated clinical course compared with Tet2+/+ mice. To continue addressing my hypothesis, I will first examine the impact of TET2 deficiency in myeloid cells on clinical and pathological outcomes of EAE (Aim 1). I will do this by examining the EAE disease course, pathology, and cellular infiltration in mice where Tet2 is knocked out specifically in myeloid cells using a Cre/lox genetic system. Additionally, I will identify the targets of TET2 in CNS-infiltrating myeloid cells, such as the suppression of pro-inflammatory cytokine production and the conversion of myeloid cells from a pro-inflammatory phenotype to an anti-inflammatory phenotype (Aim 2). Completion of these aims will contribute to the fields of neuroimmunology and epigenetics by exploring a novel mechanism of myeloid cell regulation during CNS autoimmunity, and may lead to the discovery of novel myeloid cell-specific therapeutic targets for the treatment of MS and related disorders.

项目摘要 多发性硬化症（MS）是中枢神经系统（CNS）的炎症性脱髓鞘疾病 在病因中自身免疫。 MS病变的分析以及实验自身免疫性的研究 脑脊髓炎（EAE）是MS的鼠模型，表明髓样细胞可能是关键的成分 中枢神经系统自身免疫的发病机理。最近，将十个易位酶2（TET2）确定为 MS的遗传敏感性基因座。 TET2是Fe（III），α-酮戊二酸依赖性酶，可催化催化 将甲基化的胞嘧啶氧化为5'-羟基甲基胞嘧啶（5HMC），并促进活性脱甲基化。 5hmc 是一个稳定的表观遗传标记，可以激活或反映细胞和局部依赖性的基因表达 方式。越来越多的证据表明TET2充当髓样细胞激活和功能的负调节剂， 促进稳态或抗炎细胞状态。我的假设是TET2限制了 髓样细胞的炎症功能限制CNS期间炎症，神经系统损害和残疾 自身免疫性。支持这一假设，发现TET2和5HMC在外周血中降低 与对照组相比，MS患者的单核细胞。我们实验室的初步数据 证明外周血单核细胞中TET2和5HMC的降低在 EAE，在CNS浸润的髓样细胞中，TET2转录本和5HMC含量将进一步降低 与外围。另外，将脑源性Th17细胞自适应转移到TET2 +/-小鼠中 与TET2+/+小鼠相比，临床病程恶化。为了继续解决我的假设，我将首先 检查TET2缺乏对髓样细胞对EAE临床和病理结果的影响（AIM 1）。我 将通过检查TET2为小鼠的EAE病病程，病理和细胞浸润来做到这一点 使用CRE/LOX遗传系统在髓样细胞中专门拆除。此外，我将确定 CNS浸润的髓样细胞中的TET2，例如抑制促炎性细胞因子的产生和 髓样细胞从促炎的表型转化为抗炎表型（AIM 2）。 这些目标的完成将通过探索小说来为神经免疫学和表观遗传学的领域做出贡献 CNS自身免疫期间髓样细胞调节的机制，可能导致新型髓样 细胞特异性治疗靶标，用于治疗MS和相关疾病。