Role of dysregulated miRNA in Tregs in Multiple Sclerosis

Tregs 中失调的 miRNA 在多发性硬化症中的作用

• 批准号: 8463637
• 负责人: Amy E Lovett-Racke
• 金额: $ 18.4万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-10-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8463637
• 关键词: 3' Untranslated Regions; Affect; Algorithms; Autoimmune Diseases; Biological Assay; CD4 Positive T Lymphocytes; Cell Differentiation process; Cells; Characteristics; Defect; Development; Disease; Disease Progression; Disease susceptibility; Environment; Equilibrium; Figs - dietary; Functional disorder; Gene Expression; Gene Targeting; Genes; Health; Human; Immune; Immune system; Inflammatory; Laboratories; Luciferases; Maintenance; Mediating; MicroRNAs; Multiple Sclerosis; Nerve Degeneration; Neurologic; Pathology; Pathway interactions; Patients; Phenotype; Play; Predisposition; Production; Proteins; RNA Interference Pathway; Regulatory T-Lymphocyte; Relative (related person); Role; Signal Pathway; Signal Transduction; Sorting - Cell Movement; T-Lymphocyte; Th1 Cells; Th2 Cells; Work; central nervous system demyelinating disorder; cytokine; disability; in vivo; inhibitor/antagonist; innovation; memory CD4 T lymphocyte; novel; peripheral blood; receptor; young adult

DESCRIPTION (provided by applicant): Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the central nervous system (CNS) which can result in severe neurological deficits. The cause of the disease is unknown, the immune and neurodegenerative mechanisms underlying the pathophysiology of the disease are poorly understood, and current therapies are only partially effective at slowing disease progression. My laboratory has performed a large miRNA profiling study on naive and effector/memory CD4 T cell in untreated MS patients to determine if miRNA may contribute to disease susceptibility or progression. The TGF¿ signaling pathway was predicted to be suppressed in MS patients by 12 miRNAs identified as being dysregulated in CD4 T cells. TGF¿ has shown to play a vital role in CD4+CD25hiFoxp3+ regulatory T cells (Tregs), and Tregs have been shown to be defective in MS patients. TGF¿ signaling has been shown to be critical for production of iTregs, and the maintenance of suppressor functions and Foxp3 expression in Tregs. We hypothesize that dysregulated miRNA expression in CD4 T cells in MS patients' results in impaired TGF¿ signaling and defective CD4 regulatory T cells, promoting the onset and progression of MS. Aim 1: To determine if miRNAs identified as dysregulated in CD4 T cells in MS patients directly target specific molecules in the TGF ¿ signaling pathway. First, we will determine if there is a correlation in gene expression levels between specific miRNAs and predicted TGF¿ targets in human CD4 T cells from MS patients and healthy controls. Second, we will use luciferase assays to determine if specific miRNAs suppress the 3' UTR sequence of the predicted target genes in the TGF¿ pathway. Aim 2: To determine if the miRNAs that target the TGF¿ pathway alter iTreg development and/or suppressive functions of Tregs. TGF¿ targeting miRNAs will be inhibited or over-expressed in naive CD4 T cells and then cultured under iTreg conditions. The resulting T cells will be phenotyped for iTreg characteristics. In addition, Tregs will be transfected with specific miRNA inhibitors, evaluated for sensitivity to TGF¿, and ability to suppress effector T cells. Aim 3: To determine if there is an association between the level of TGF¿-associated miRNA expression and the number of Tregs in vivo, or an association between TGF¿-associated miRNA expression and suppressive capacity of Tregs in MS patients. Flow cytometric analysis of peripheral blood CD4 T cells from MS patients will evaluate the number of Tregs in MS patients relative to the expression of TGF¿-associated miRNAs. In addition, Tregs will be sorted from peripheral blood of MS patients, evaluated for suppressive capacity, and determine if there is a correlation between TGF¿-associated miRNAs and Treg number and/or function. Given the well-documented role of Tregs in maintaining tolerance to self and the observation that CD4 Tregs are defective in MS patients, this novel study will determine if dysregulated miRNA expression in CD4 T cells underlies this Treg defect in MS patients. Although several studies have identified miRNAs that play a role in Tregs17-22, no studies have identified differentially expressed miRNAs in MS patients that are specifically associated with genes in the TG¿ pathway and their influence on Tregs in an autoimmune disease. Treg cells play an important role in the delicate balance of protection from our environment and protecting ourselves from our own immune system, and TGF¿ has pleiotrophic effects that extend beyond Tregs that have vast importance in health and disease, making this an invaluable study.

描述（由申请人提供）：多重扇形（MS）是中枢神经系统（CNS）的脱髓鞘疾病，导致该疾病的原因尚不清楚。疾病对保守党的疾病对未经治疗的MS患者的天真和效应子/记忆CD4 T细胞进行了较大的miRNA分析，以确定miRNA是否有助于ISEASE敏感性或TGF。预计在CD4 T细胞中被鉴定为功能障碍的12个miRNA被预测在MS患者中被抑制。已显示在CD4+ CD25HIFOXP3+常规性T细胞（TREG）中起着至关重要的作用，并且Tregs已显示在某些患者中是有缺陷的。信号传导对于生产ITREGS ZE至关重要，即MS患者的CD4 T细胞中miRNA表达失调会导致TGFS的受损。信号传导和有缺陷的CD4调节性T细胞的目的1：确定MRNASREGRENCHENE MS患者中的4个T细胞是否直接靶向TGF„中的特定分子。信号通路。来自MS患者和健康对照的人CD4 T细胞的靶标。途径2：确定miRNA是否针对TGF？途径改变了TGFS的ITREG开发和/或抑制功能。靶向miRNA，在幼稚的CD4 T细胞中表达，然后将其培养为ITREG特征。和抑制效果t -cells的能力。 - 相关的miRNA表达和体内Treg的数量，或TGF¿之间的关联 - 在MS患者中相关的miRNA表达和Tregs的抑制能力。 - 相关的miRNA。此外，Treg将从MS患者的血液中排序 - 相关的miRNA和TREG数量和或功能在MS患者中有缺陷，如果CD4 T细胞中的miRNA表达失调，则在MS患者中表达了MIRNA，而MIRNA缺陷。 tg ??途径和对自身免疫性疾病中Treg的影响。具有多余的毒素作用，超越了在健康和疾病中具有广泛重要性的Treg之外，这是一项非常宝贵的研究。