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

基本信息

批准号：
9339545
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
项目类别：
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/9339545
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 Cytokine Gene Cytoprotection DNA Fragmentation Data Demyelinating Diseases Dioxygenases Disease Disease remission Enzyme-Linked Immunosorbent Assay Experimental Autoimmune Encephalomyelitis Fatigue Flow Cytometry Functional disorder GATA3 gene Gene Expression Genetic Transcription Goals 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 Neuraxis 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 T-Cell Activation T-Lymphocyte TNF gene TdT-Mediated dUTP Nick End Labeling Assay Testing Therapeutic Transcription Alteration Umbilical Cord Blood Up-Regulation Veterans Visual Water Western Blotting autoreactive T cell autoreactivity base calpain inhibitor chemokine cytokine immunoregulation indoleamine interleukin-23 monocyte multiple sclerosis patient neuron loss neuroprotection neutralizing antibody novel novel therapeutics public health relevance release factor therapy development transcription factor

项目摘要

DESCRIPTION (provided by applicant): 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, NFkappaB). 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 identfy the mechanisms by which calpain inhibitor therapy may potentially treat MS.

描述（由申请人提供）：多发性硬化症 (MS) 是一种与中枢神经系统神经变性相关的脱髓鞘疾病，被认为是由自身反应性 T 辅助 (Th) 细胞攻击髓磷脂蛋白引起的。复发缓解型 (RR) MS 的特点是急性发作，发作之间有部分或完全恢复期。目前的研究结果表明，促炎性 Th1 和 Th17 细胞在复发期间占主导地位，而抗炎性 Th2 和 T 调节细胞 (Treg) 则推动缓解。了解这些转变的机制对于开发多发性硬化症的治疗方法至关重要。此前，我们的数据显示，与缓解期相比，多发性硬化症患者复发期间大脑、脊髓和外周血单核细胞 (PBMC) 中钙激活蛋白酶钙蛋白酶的表达和活性有所增加。钙蛋白酶激活 T 细胞并参与 Th1 细胞因子的产生，而其抑制作用则促进 Th2 细胞因子的上调。钙蛋白酶还调节与 Th 谱相关的信号蛋白（STAT、NFAT、NFkappaB）的活性。我们的目标是确定多发性硬化症患者复发和缓解期间钙蛋白酶活性的改变是否通过转录因子的改变与 Th1/Th2 失调有关。初步数据发现，从 RR-MS 患者血液中分离的 PBMC 中 IL-17 水平升高，用 calpeptin 处理这些细胞可减少 MBP 激活的 PBMC 中 Th17 细胞的数量，并抑制 MBP 特异性 T 细胞的增殖。钙蛋白酶抑制还增加了 MS PBMC 中骨髓源性抑制细胞 (MDSC) 的数量。有趣的是，特异性诱导 Th2 细胞因子基因表达的 STAT6 在 MS 患者的活化 PBMC 中被降解，并且与钙蛋白酶抑制剂一起孵育可逆转降解，表明钙蛋白酶抑制可能通过减少 STAT6 降解来促进 Th2 谱。将原代神经元培养物与从患者体内分离出的活化 PBMC 的上清液一起培养会增加神经元死亡，但钙蛋白酶抑制剂治疗可阻止这种死亡。基于这些数据，我们假设钙蛋白酶抑制通过改变多发性硬化症中的转录因子和骨髓源性抑制细胞而使 Th 细胞失调，并减少释放到 Th1/Th17 细胞上清液中诱导细胞死亡的炎症介质的产生。将使用三个具体目标来检验该假设：（1）检验钙蛋白酶抑制对 MS 患者中 Th 和 Treg 细胞因子/趋化因子谱以及 MDSC 改变的影响； (2) 确定钙蛋白酶抑制后导致这些变化的信号传导机制； (3) 确定活化的 Th 细胞分泌的哪些细胞因子/趋化因子导致细胞死亡或增加体外对神经元的保护，并检查钙蛋白酶抑制对这些因子分泌的影响。这些研究对于确定钙蛋白酶抑制剂疗法可能治疗多发性硬化症的机制至关重要。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Distinct Cytokine and Chemokine Expression in Plasma and Calpeptin-Treated PBMCs of a Relapsing-Remitting Multiple Sclerosis Patient: A Case Report.

DOI：
10.1007/s11064-018-2655-z
发表时间：
2018-12
期刊：
NEUROCHEMICAL RESEARCH
影响因子：
4.4
作者：
Chandran, Raghavendar;Capone, Mollie;Matzelle, Denise;Polcyn, Rachel;Kau, Elizabeth;Haque, Azizul;Banik, Naren L.
通讯作者：
Banik, Naren L.

Cytokine/chemokine dysregulation in progressive MS patient is apparent and can be modulated by calpain inhibition.