Regulation of inflammatory T Cells and Neuroprotection by Calpain Inhibitor in MS

多发性硬化症中钙蛋白酶抑制剂对炎症 T 细胞的调节和神经保护作用

• 批准号: 8842002
• 负责人: NAREN L BANIK
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8842002
• 关键词: Acute; Anti-Inflammatory Agents; Anti-inflammatory; Blood; Brain; CCL2 gene; CD14 gene; CD28 gene; CD3 Antigens; CD4 Positive T Lymphocytes; Calpain; Caspase; Cell Culture Techniques; Cell Death; Cells; Cessation of life; Cytokine Gene; Cytoprotection; DNA Fragmentation; Data; Demyelinating Diseases; Dioxygenases; Disease; Disease remission; Enzyme-Linked Immunosorbent Assay; Experimental Autoimmune Encephalomyelitis; Fatigue; Flow Cytometry; Functional disorder; Gene Expression; Genetic Transcription; Goals; Health; Helper-Inducer T-Lymphocyte; Human; IL8 gene; Immunohistochemistry; In Vitro; Incubated; Individual; Inflammation; Inflammation Mediators; Inflammatory; Interferons; Interleukin-10; Interleukin-13; Interleukin-17; Interleukin-2; Interleukin-4; Interleukin-5; Laboratories; Lead; Measures; Mediating; Mediation; Multiple Sclerosis; Mus; Myelin Proteins; Myelogenous; Nerve Degeneration; Neuroglia; Neurons; Oligodendroglia; Pain; Paralysed; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Population; Production; Publishing; Recovery; Regulation; Regulatory T-Lymphocyte; Relapse; Relapsing-Remitting Multiple Sclerosis; Role; STAT6 gene; Sampling; Signal Transduction; Signaling Protein; Spinal Cord; Suppressor-Effector T-Lymphocytes; System; T-Cell Activation; T-Lymphocyte; TNF gene; TdT-Mediated dUTP Nick End Labeling Assay; Testing; Therapeutic; Up-Regulation; Veterans; Visual; Water; Western Blotting; base; calpain inhibitor; chemokine; cytokine; indoleamine; interleukin-23; neuroprotection; neutralizing antibody; novel; novel therapeutics; release factor; therapy development; transcription factor

Multiple sclerosis (MS), a demyelinating disease associated with neurodegeneration of the CNS, is thought to result from an attack on myelin proteins by autoreactive T helper (Th) cells. Relapsing-remitting (RR) MS is characterized by acute attacks with periods of partial or full recovery between episodes. Current findings suggest that pro-inflammatory Th1 and Th17 cells predominate during relapse and that anti-inflammatory Th2 and T regulatory cells (Tregs) drive remission. Understanding the mechanisms of these shifts is crucial to developing therapies for MS. Previously, our data showed increased expression and activity of the calcium- activated protease calpain in brain, spinal cord, and peripheral blood mononuclear cells (PBMCs) of MS patients during relapse compared to remission. Calpain activates T cells and is involved in the production of Th1 cytokines, while its inhibition promotes upregulation of Th2 cytokines. Calpain also modulates the activity of signaling proteins associated with Th profiles (STATs, NFAT, NF¿B). Our goal is to determine whether alterations in calpain activity during relapse and remission in MS patients are involved in Th1/Th2 dysregulation via alterations in transcription factors. Preliminary data found increased IL-17 levels in PBMCs isolated from the blood of RR-MS patients, and treating these cells with calpeptin reduced the number of Th17 cells in MBP-activated PBMCs, and inhibited the proliferation of MBP-specific T cells as well. Calpain inhibition also increased the number of myeloid-derived suppressor cells (MDSC) in MS PBMCs. Interestingly, STAT6, which specifically induces Th2 cytokine gene expression, was degraded in activated PBMCs from MS patients, and incubation with calpain inhibitor reversed the degradation, suggesting that calpain inhibition may promote a Th2 profile by reducing STAT6 degradation. Incubating primary neuron cultures with supernatant from activated PBMCs isolated from patients increased neuronal death, which was blocked by calpain inhibitor treatment. Based on these data, we hypothesize that calpain inhibition dysregulates Th cells via alterations of transcription factors and myeloid-derived suppressor cells in MS, and reduces production of inflammatory mediators released into the supernatant of Th1/Th17 cells that induce cell death. Three specific aims will be used to examine the hypothesis: (1) examine the effects of calpain inhibition on Th and Treg cytokine/chemokine profiles and alteration of MDSCs in MS patients; (2) determine the signaling mechanisms responsible for these shifts following calpain inhibition; and (3) identify which cytokines/chemokines secreted by activated Th cells are responsible for cell death or increase protection of neurons in vitro and examine the effects of calpain inhibition on the secretion of these factors. These studies are crucial to identify the mechanisms by which calpain inhibitor therapy may potentially treat MS.

多发性硬化症（MS）是一种与中枢神经系统神经退行性变相关的脱髓鞘疾病