Neuronal antigen surveillance and autoimmunity in CNS demyelinating disease

中枢神经系统脱髓鞘疾病的神经元抗原监测和自身免疫

基本信息

批准号：
10380683
负责人：
Charles Lee Howe
金额：
$ 59.47万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10380683
关键词：
Acute Address Adoptive Transfer Antigen Presentation Antigen Presentation Pathway Antigen-Presenting Cells Antigens Autoimmunity Autologous Automobile Driving Axon Blood Brain CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes Cells Clone Cells Coupled Cuprizone Defect Demyelinating Diseases Demyelinations Disease Disease Progression Disease remission Effector Cell Engineering Exhibits Experimental Autoimmune Encephalomyelitis Fibroblasts Genes Goals Health Histocompatibility Antigens Class I Human Immunization Induced pluripotent stem cell derived neurons Infiltration Inflammation Injury Interferons Intervention Ion Channel Knowledge Lymphocyte MHC Class I Genes Measures Mediating Mediator of activation protein Microfluidics Mission Modeling Molecular Multiple Sclerosis Multiple Sclerosis Lesions Mus National Institute of Neurological Disorders and Stroke Nervous System Physiology Neurons OVA-8 Ovalbumin Pathogenesis Pathogenicity Patients Peptides Peripheral Population Public Health Relapse Research Resolution Role Secondary to Specificity Symptoms T-Lymphocyte Testing Time Toxic effect Transgenic Organisms Tumor-infiltrating immune cells United States National Institutes of Health Work adeno-associated viral vector axon injury base burden of illness cytotoxic CD8 T cells experimental study functional disability improved innovation loss of function multiple sclerosis patient neoantigens neuroinflammation novel novel therapeutic intervention oligodendrocyte-myelin glycoprotein prevent recruit remyelination response targeted treatment therapy development trafficking

项目摘要

There is a critical unmet need to identify the pathogenic mechanisms that drive disease progression in patients with multiple sclerosis (MS). Accumulation of axon injury and functional disability in MS are not adequately impacted by current therapies. The long-term goal of this work is to discover new strategies to prevent or reverse disease progression in MS. Despite evidence that CD8+ T cells are associated with axon injury and progression in MS, the functional role for these cells and the relevant mechanisms required for recruitment of these cells to the demyelinated brain are unknown. The antigenic targets of neuron-specific CD8+ T cells are also unknown. The overall objectives of this study are to test the mechanistic role of neuron antigen-specific CD8+ T cells in the injury of demyelinated axons and to determine whether patients with MS have such cells. The rationale is that axon injury is the primary substrate of progression in MS, axonal MHC class I expression is upregulated by inflammation and demyelination, and cytotoxic CD8+ T cells directed against neuron-specific antigens injure demyelinated axons. The central hypothesis of this proposal is that neuron antigen-specific CD8+ T cells injure demyelinated axons. Guided by strong preliminary evidence, the hypothesis will be tested using AAV-mediated transduction of neurons to drive expression of the neoantigen ovalbumin (OVA) within the context of CNS demyelination induced by cuprizone toxicity or immunization against a myelin oligodendrocyte glycoprotein-derived peptide (MOG-EAE) in hosts that have transgenic CD8+ T cells directed against the OVA- derived peptide SIINFEKL (OT-I). The study will also use autologous T cells and fibroblast-derived iPSC- derived neurons grown in microfluidic chambers to determine whether MS patient CD8+ T cells injure their own axons. Three specific aims will be pursued: 1) determine the mechanisms of CD8+ T cell-mediated axon injury in the demyelinated CNS; 2) identify the cellular locus of MHC class I expression required for axon injury and determine how demyelination drives CNS infiltration of neuron antigen-specific CD8+ T cells; 3) determine whether MS patients have neuron-antigen specific CD8+ T cells. This approach is conceptually innovative because of the proposal that demyelination and inflammation induce axonal presentation of self-peptides on MHC class I. The approach is technically innovative based on the use of novel AAV vectors to drive neoantigens in neurons within the demyelinated CNS, selective deletion of brain-resident antigen presenting cells (APCs) vs peripheral APCs, use of multiple host manipulations coupled to adoptive transfer of traceable effector cells to temporally and spatially profile anti-neuronal T cell trafficking, and the use of patient-derived neurons and autologous CD8+ T cells. This work will make a significant, powerful impact on the field by revealing the capacity of CD8+ T cells directed against neuron-specific antigens to injure demyelinated axons and by identifying such T cells in patients with MS.

确定驱动患者疾病进展的致病机制有至关重要的需求多发性硬化症（MS）。 MS中轴突损伤和功能障碍的积累不足受当前疗法的影响。这项工作的长期目标是发现预防或 MS的反向疾病进展。尽管有证据表明CD8+ T细胞与轴突损伤和 MS的进展，这些细胞的功能作用以及募集所需的相关机制这些细胞到脱髓鞘的大脑尚不清楚。神经元特异性CD8+ T细胞的抗原靶标是也未知。这项研究的总体目标是测试神经元特异性的机械作用 CD8+ T细胞在脱髓鞘轴突损伤中，并确定MS患者是否具有此类细胞。理由是，轴突损伤是MS，轴突MHC I类表达的主要基质炎症和脱髓鞘和针对神经元特异性的细胞毒性CD8+ T细胞上调抗原损伤脱髓鞘的轴突。该提议的中心假设是神经元抗原特异性 CD8+ T细胞会损害脱髓鞘的轴突。在有力的初步证据的指导下，将检验该假设使用AAV介导的神经元转导驱动新抗原卵蛋白（OVA）的表达 CNS脱髓鞘的背景是由丘陵毒性毒性或免疫对髓磷脂少突胶质细胞引起的具有针对OVA-的转基因CD8+ T细胞的宿主中糖蛋白衍生的肽（MOG-EAE）衍生的肽Siinfekl（OT-I）。该研究还将使用自体T细胞和成纤维细胞衍生的IPSC- 在微流体室中生长的神经元，以确定MS患者CD8+ T细胞是否伤害自己轴突。将追求三个具体目标：1）确定CD8+ T细胞介导的轴突损伤的机制在脱髓鞘的中枢神经系统中； 2）确定轴突损伤所需的MHC I类表达的细胞基因座和确定脱髓鞘如何驱动神经元特异性CD8+ T细胞的CNS渗透； 3）确定 MS患者是否患有神经元特异性CD8+ T细胞。这种方法在概念上是创新的由于提出的脱髓鞘和炎症会诱导自肽的轴突表现 MHC类I。该方法在技术上是基于使用新型AAV矢量驱动的创新性脱髓鞘中枢神经系统内神经元中的新抗原，选择性删除脑居民抗原呈现单元格（APC）与外围APC，使用多个宿主操纵，并结合可追溯的产物传递效应细胞在时间和空间上介绍抗神经元T细胞运输，并使用患者衍生神经元和自体CD8+ T细胞。这项工作将对该领域产生重大，强大的影响揭示针对神经元特异性抗原的CD8+ T细胞的能力损害脱髓鞘轴突并通过鉴定MS患者中的T细胞。