Role of miRNA Dysregulation on T Cell Differentiation and Function in MS

miRNA 失调对 MS 中 T 细胞分化和功能的作用

• 批准号: 10551306
• 负责人: Amy E Lovett-Racke
• 金额: $ 44.12万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-03 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10551306
• 关键词: Affect; Autoimmune; Autoimmune Diseases; CD4 Positive T Lymphocytes; CNS autoimmunity; Cell Differentiation process; Cell physiology; Cells; Complement; Data; Defect; Development; Disease; Disease Progression; Disease susceptibility; Experimental Autoimmune Encephalomyelitis; Failure; Functional disorder; Gene Expression; Genes; Human; Immune; In Vitro; Individual; Inflammatory; Interleukin-6; Laboratories; Mediating; MicroRNAs; Modeling; Multiple Sclerosis; Mus; Nerve Degeneration; Neurologic Deficit; Pathology; Pathway interactions; Persons; Play; Population; Predisposition; Proteins; RNA Interference Pathway; Receptor Signaling; Regulatory T-Lymphocyte; Risk; Risk Factors; Role; Signal Pathway; Signal Transduction; T cell differentiation; T-Lymphocyte; Testing; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; autoreactive T cell; central nervous system demyelinating disorder; cytokine; disability; experimental study; in vivo; inhibitor; innovation; memory CD4 T lymphocyte; mouse model; multiple sclerosis patient; novel; overexpression; prevent; therapeutic miRNA; therapeutic target; young adult

Abstract 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. There is a tremendous need to investigate novel mechanisms that may be contributing to disease susceptibility and the pathophysiology of this disease. To this end, my laboratory has performed a large miRNA profiling study on naïve and effector/memory CD4 T cell in untreated MS patients to determine if miRNA may contribute to disease susceptibility or progression. MiRNA negatively regulate gene expression via the RNA interference pathway, and thus play a key role modulating the level of specific proteins in cells. We have identified at least two pathways that are altered in MS patients that may contribute to their susceptibility to develop MS. First, miRNAs targeting components of the Th2 cell differentiation pathway were elevated in MS patients T cells, skewing differentiation into pro-inflammatory Th1 cells. Second, miRNAs targeting the TGFβ signaling pathway limited the differentiation of regulatory T cells. Thus, the defects observed in MS patients CD4 T cells may be at least partially mediated by miRNA dysregulation. In this proposal, we will address the following questions. Aim 1: Are MS patients’ CD4 T cells defective in their ability to differentiate into Tregs in a miRNA-dependent manner? Preliminary data indicates that naïve CD4 T cells from MS patients fail to efficiently differentiate into Tregs. Using miRNA inhibitors, we will determine if this failure to differentiate into Tregs is dependent on specific miRNAs over-expressed in MS patients and whether Treg differentiation can be normalized MS patients’ naïve CD4 T cells. Aim 2: How does over-expression of MS-associated miRNAs affect the development and progression of CNS autoimmunity in a mouse model? Using EAE, the role of miRNAs that target CD4 T cell differentiation into effector and regulatory T cells will be analyzed in vivo to complement the human in vitro experiments. Aim 3: Can expression level of miR-128, which targets both effector and regulatory CD4+ T cells, modulate the risk of CNS autoimmunity? We found that miR-128 targets proteins in the Th2 and TGFβ signaling pathways, promoting the differentiation of Th1 cells and preventing the development of Tregs. Using CD4-specific miR-128-/- mice and mice overexpressing miR-128 in CD4 T cells, we will determine if loss of miR-128 in CD4 T cells minimizes susceptibility to CNS autoimmunity, normalizes CD4 T cell differentiation into effector and regulatory cells, and thus, is a potential therapeutic target to correct both effector and regulatory T cell defects in MS. In contrast, we will use mice overexpressing miR-128 in CD4 T cells to determine if miR-128 is sufficient to skew CD4 T cell differentiation and enhance the risk of developing CNS autoimmunity. This study will test the hypothesis that miRNA dysregulation in naïve CD4+ T cells is an underlying risk factor in CNS autoimmunity that can be therapeutically targeted to normalize effector and regulatory CD4 T cell function. If our hypothesis is correct, miRNA-based therapies may not only prevent CNS autoimmunity in susceptible individuals, but ameliorate autoreactive T cells in patients with MS.

抽象的 多发性硬化症（MS）是一种免疫介导的中枢神经系统（CNS）的脱髓鞘疾病 在严重的神经系统缺陷中。该疾病的原因尚不清楚，免疫和神经退行性机制 该疾病的病理生理学的基本知识很少，目前的疗法仅部分有效 疾病进展减慢。需要研究可能有助于的新型机制 疾病的敏感性和该疾病的病理生理学。为此，我的实验室进行了大型miRNA 对未治疗的MS患者的幼稚和效应子/记忆CD4 T细胞的分析研究，以确定miRNA是否可以贡献 miRNA通过RNA干扰途径负调节基因表达， 因此，起着调节细胞中特定蛋白质水平的关键作用。我们已经确定了至少两个途径 MS患者可能会改变其发展MS的敏感性。首先，靶向组件的miRNA MS患者T细胞中Th2细胞分化途径的升高，偏分化为促炎性 Th1细胞。其次，靶向TGFβ信号通路的miRNA限制了调节T细胞的分化。那， 在MS患者中观察到的缺陷CD4 T细胞可能至少部分通过miRNA失调介导。在此提案中， 我们将解决以下问题。 AIM 1：MS患者的CD4 T细胞是否有缺陷 以miRNA依赖性方式进入Treg？初步数据表明，来自MS患者的幼稚CD4 T细胞无法 有效地分化为Tregs。使用miRNA抑制剂，我们将确定这种未分化为tregs是否为 取决于MS患者过表达的特定miRNA，以及是否可以将Treg分化归一化ms 患者的幼稚CD4 T细胞。目标2：MS相关的miRNA的过表达如何影响发展和 小鼠模型中CNS自身免疫性的进展？使用EAE，靶向CD4 T细胞分化的miRNA的作用 将在效应子和调节性T细胞中进行体内分析，以完成人体体外实验。目标3：可以 miR-128的表达水平靶向效应子和调节性CD4+ T细胞，调节CNS的风险 自身免疫性？我们发现miR-128靶向TH2和TGFβ信号通路中的蛋白质，从而促进 Th1细胞的分化并防止Treg的发展。使用CD4特异性miR-128 - / - 小鼠和小鼠 在CD4 T细胞中过表达miR-128，我们将确定CD4 T细胞中miR-128的损失是否使对CNS的敏感性最小 自身免疫性将CD4 T细胞分化为效应细胞和调节细胞，因此是一种潜在的治疗 纠正MS中效应子和调节性T细胞缺陷的目标。相比之下，我们将使用过表达miR-128的小鼠 在CD4 T细胞中，以确定miR-128是否足以使CD4 T细胞分化并增强发展的风险 CNS自身免疫。这项研究将检验以下假设：幼稚的CD4+ T细胞中的miRNA失调是一种基础 CNS自身免疫性中的危险因素可以被热针对效应子和调节性CD4 T细胞归一化 功能。如果我们的假设正确，基于miRNA的疗法不仅可以防止CNS自身免疫性 个体，但可以改善MS患者的自动反应性T细胞。