Sex-specific genomic mechanisms of transcriptional regulation in ME/CFS/SEID

 描述（由申请方提供）：肌痛性脑脊髓炎/慢性疲劳综合征/全身性呼吸不耐受病（ME/CFS/SEID）的当前管理仅依赖于症状管理来改善生活质量，但未解决疾病发作和进展的潜在机制。为了更好地了解ME/CFS/SEID呈现所涉及的关键生物学靶标，本研究提案的主要目标是确定ME/CFS/SEID的男性特异性生物标志物和治疗靶标，并提供性别特异性疾病发病和进展的见解，这将导致更好的治疗干预。本研究提案旨在使用来自我们最近NIH资助的研究（1 R 01 NS 090200 -01（PI弗莱彻））招募的患者的外周血单核细胞（PBMC）。这些PBMC已在三个时间点从男性ME/CFS/SEID患者和匹配的健康对照中分离出来-在运动激发前、在最大努力（VO 2 max）时和在最大努力后四小时。本提案的具体目标包括：确定代谢途径和可能的调节RNA影响男性ME/CFS/SEID。我们的假设是，未知的成绩单和选择性剪接事件调节的基因表达的变化，负责炎症反应，白细胞迁移，调节细胞发育。我们的目标是使用RNA-seq方法来鉴定已知和新型转录物的丰度，并在男性患者的PBMC中发现新的剪接事件。这将允许建立一组性别特异性ME/CFS/SEID候选转录本。然后，我们将使用NanoString Technologies的nCounter系统验证它们，并创建一组转录本，可能用于ME/CFS的诊断和管理。 具体目标2。研究ME/CFS/SEID患者转录调控的可能机制。为了更好地了解ME/CFS/SEID男性患者基因表达变化的原因，我们的目标是确定拷贝数变异（CNV）和基因组DNA甲基化的变化。我们将使用Agilent SurePrint G3人类基因组CGH+SNP微阵列来鉴定一组ME/CFS/SEID性别特异性候选CNV。我们将使用Illumina Infinium Human Methylation 450 BeadChip微阵列来评估ME/CFS/SEID患者和健康对照之间甲基化模式的差异。 具体目标3。比较受影响的代谢途径和转录调控机制的男性和女性ME/CFS。我们将比较男性ME/CFS/SEID患者和HC患者之间的差异，以及女性中的相同差异（来自最近资助的赠款提案1 R15 NS 087604 - 01 A1，PI Nathanson）。将对基因表达、选择性剪接、基因组DNA甲基化模式和CNVs的结果进行比较。我们期望发现参与炎症反应和免疫系统过程的基因调控的差异，特别是在细胞因子产生和活性的调控方面。

 DESCRIPTION (provided by applicant): Current management of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome/Systemic Exertion Intolerance Disease (ME/CFS/SEID) relies solely on symptom management to improve quality of life but does not address the underlying mechanisms of disease onset and progression. To provide better insight into the key biological targets involved ME/CFS/SEID presentation, the main objective of this research proposal is to identify male-specific biomarkers and therapeutic targets of ME/CFS/SEID and provide insight into sex-specific disease onset and progression, which will lead to the better therapeutic intervention. This research proposal aims to use Peripheral Blood Mononuclear Cells (PBMC) from patients recruited for our recently NIH funded research (1R01NS090200-01 (PI Fletcher)). These PBMC have been isolated from male ME/CFS/SEID patients and matched healthy controls at three time points - before exercise challenge, at the peak of effort (VO2 max) and four hours after the peak effort. Specific Aims of this proposal include: Specific Aim 1. Identify metabolic pathways and possible regulatory RNAs affected in men with ME/CFS/SEID. Our hypothesis is that unknown transcripts and alternative splicing events regulate changes in expression of genes responsible for inflammatory response, leukocyte migration, and regulation of cell development. We aim to use an RNA-seq approach to identify the abundance of known and novel transcripts and discover new splicing events in PBMC of male patients. This will allow establishing a set of sex-specific ME/CFS/SEID candidate transcripts. We will then validate them using NanoString Technologies' nCounter system and create a panel of transcripts to potentially be used in the diagnosis and management of ME/CFS. Specific Aim 2. To investigate possible mechanisms of transcriptional regulation in men with ME/CFS/SEID. To better understand causes of changes in gene expression in male patients with ME/CFS/SEID, we aim to identify changes in copy number variation (CNV) and genomic DNA methylation. We will use Agilent SurePrint G3 Human Genome CGH+SNP microarrays to identify a set of ME/CFS/SEID sex specific candidate CNVs. We will use Illumina Infinium Human Methylation450 BeadChip microarrays to evaluate differences in the methylation patterns between ME/CFS/SEID patients and healthy controls. Specific Aim 3. Compare affected metabolic pathways and mechanisms of transcriptional regulation in men and women with ME/CFS. We will compare differences between ME/CFS/SEID patients and HC in men with the same differences in women (from the recently funded grant proposal 1 R15 NS087604-01A1, PI Nathanson). Comparisons will be done for the results in gene expression, alternative splicing, patterns on the genomic DNA methylation and CNVs. We expect to find differences in the regulation of genes involved in inflammatory response and immune system processes, particularly in the regulation of cytokines production and activity.