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：
10213156
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/10213156
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是一个尚未得到满足的关键需求患有多发性硬化症（MS）。多发性硬化症中轴突损伤和功能障碍的积累没有得到充分的解决受当前疗法的影响。这项工作的长期目标是发现新的策略来预防或逆转多发性硬化症的疾病进展。尽管有证据表明 CD8+ T 细胞与轴突损伤有关 MS 的进展、这些细胞的功能作用以及招募这些细胞对脱髓鞘大脑的作用尚不清楚。神经元特异性 CD8+ T 细胞的抗原靶点是亦未知。本研究的总体目标是测试神经元抗原特异性的机制作用 CD8+ T 细胞在脱髓鞘轴突损伤中的作用，并确定 MS 患者是否有此类细胞。基本原理是轴突损伤是 MS 进展的主要基础，轴突 MHC I 类表达炎症和脱髓鞘作用上调，细胞毒性 CD8+ T 细胞针对神经元特异性抗原会损伤脱髓鞘的轴突。该提案的中心假设是神经元抗原特异性 CD8+ T 细胞损伤脱髓鞘轴突。在强有力的初步证据的指导下，假设将得到检验使用 AAV 介导的神经元转导来驱动新抗原卵清蛋白 (OVA) 的表达铜宗毒性或针对髓磷脂少突胶质细胞的免疫诱导的中枢神经系统脱髓鞘的背景糖蛋白衍生肽 (MOG-EAE) 在具有针对 OVA- 的转基因 CD8+ T 细胞的宿主中衍生肽 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衍生神经元，以确定 MS 患者 CD8+ T 细胞是否损伤自身轴突。我们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 类表达的细胞位点确定脱髓鞘如何驱动神经元抗原特异性 CD8+ T 细胞的中枢神经系统浸润； 3）确定 MS 患者是否具有神经元抗原特异性 CD8+ T 细胞。这种方法在概念上是创新的因为脱髓鞘和炎症会诱导自肽的轴突呈递 MHC I 类。该方法在技术上是创新的，基于使用新型 AAV 载体来驱动脱髓鞘中枢神经系统神经元中的新抗原，选择性删除大脑驻留抗原呈递细胞（APC）与外围 APC，使用多种宿主操作以及可追踪的过继转移效应细胞在时间和空间上分析抗神经元 T 细胞运输，以及使用患者来源的神经元和自体 CD8+ T 细胞。这项工作将对该领域产生重大而强大的影响揭示 CD8+ T 细胞针对神经元特异性抗原损伤脱髓鞘轴突的能力并通过鉴定多发性硬化症患者体内的此类 T 细胞。