DNA methylation in the development of multiple sclerosis

总结 本申请涉及影响多发性硬化（MS）风险的表观遗传学事件的研究 和进步。我们目前的初步结果与广泛的微分一致 在诊断时，外周血CD 19 + B细胞的低甲基化，构成了一个机制联系， 抗CD 20抗体治疗本病的临床疗效。我们在这些成果的基础上， 获得信息丰富的各种数据和样本集，在具体目标1中提出同时 评估单细胞基因表达（scRNA-seq）、染色质可及性（scATAC-seq）和 在配对的脑脊液（CSF）和外周血单核细胞的细胞表面标志物， 临床发作时初治MS患者和匹配良好的对照，并在目标2中链接至 个体的DNA变异，以发展与以下相关的全球和细胞特异性遗传负担： 疾病表达。在目标3中，我们将新兴的表观遗传和转录组特征与 使用Beacon®系统光流体平台来可视化和评估细胞表型 在单细胞水平上。在目标4中，我们实施靶向三模式单细胞测定以描述靶向三模式单细胞测定。 主要MS易感位点的表观遗传景观，主要组织相容性复合体， 染色体6p 21。通过系统地整合单细胞染色质状态、DNA变异和基因 表达数据，我们希望获得重要的新信息的发病机制。而且还要 鉴定与MS临床发作相关的细胞特异性表观遗传特征， 做作状态的生物标志物。对研究参与者进行细致的临床随访， 评估其预后潜力的机会。

Summary This application is concerned with the study of epigenetics events affecting multiple sclerosis (MS) risk and progression. We present preliminary results consistent with widespread differential hypomethylation in peripheral CD19+ B cells at the time of diagnosis, posing a mechanistic link to the clinical efficiency of anti-CD20 antibody treatment for this disease. We build on these results and access to informative and diverse data- and sample-sets to propose in Specific Aim 1 the simultaneous assessment of single-cell gene expression (scRNA-seq), chromatin accessibility (scATAC-seq), and cell surface markers in paired cerebrospinal fluid (CSF) and peripheral blood mononuclear cells from treatment naïve MS patients at the time of clinical onset and well matched controls, and link in Aim 2 to the individuals’ DNA variance to develop global and cell-specific genetic burdens associated with disease expression. In Aim 3 we connect the emerging epigenetic and transcriptome signatures with cell function using the Beacon® system optofluidic platform to visualize and assess cellular phenotypes at the single-cell level. In Aim 4 we implement a targeted trimodal single-cell assay to describe the epigenetic landscape of the principal MS susceptibility locus, the Major Histocompatibility Complex in chromosome 6p21. By systematically integrating single cell chromatin states, DNA variance, and gene expression data, we expect to gain important novel information about pathogenesis. Moreover, we will identify cell-specific epigenetic signatures associated with MS clinical onset, potentially serving as biomarkers of affectation status. The meticulous clinical follow up of study participants offers an opportunity to assess their prognostic potential.