Deciphering Early Stages of Polyomavirus CNS Pathogenesis and Immunity

破译多瘤病毒中枢神经系统发病机制和免疫的早期阶段

• 批准号: 10610484
• 负责人: Aron Eliot Lukacher
• 金额: $ 89.66万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2030-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610484
• 关键词: Acquired Immunodeficiency Syndrome; Address; Affinity; Animal Model; Autoimmune; Biological Products; Brain; Brain Diseases; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Capsid Proteins; Central Nervous System Diseases; Central Nervous System Infections; Cerebrum; Complication; Cryoelectron Microscopy; Custom; Demyelinations; Development; Disease; Ependyma; Epitopes; Event; Family; Foundations; Hematologic Neoplasms; Heterogeneity; Human; Immune; Immunity; Immunologics; Immunotherapeutic agent; Incidence; Individual; Infection; Inflammatory; Invaded; JC Virus; Lesion; Life; Link; Maintenance; Maps; Memory; Modeling; Mus; Mutation; Natural Immunity; Pathogenesis; Patients; Polyomavirus; Progressive Multifocal Leukoencephalopathy; Reaction; Resolution; STAT1 gene; Staging; T cell response; T-Lymphocyte; Therapeutic; Three-Dimensional Imaging; Tropism; Tysabri; Variant; Ventricular; Virus; Visualization; brain parenchyma; chemotherapeutic agent; chronic infection; drug withdrawal; human pathogen; image reconstruction; immune modulating agents; in vivo; in vivo evaluation; interleukin-21; mortality; mouse polyomavirus; multiple sclerosis patient; natalizumab; neuropathology; neutralizing antibody; next generation sequencing; pathogen; preservation; tissue culture

ABSTRACT JC polyomavirus (JCPyV), a ubiquitous human pathogen, causes several devastating brain diseases in immune compromised individuals. The most notable of these JCPyV-associated CNS diseases is the frequently fatal demyelinating brain disease progressive multifocal leukoencephalopathy (PML). PML, an AIDS-defining lesion in the pre-cART epoch, has emerged as a life-threatening complication in patients receiving immunomodulatory agents for autoimmune and inflammatory disorders and treatment for certain hematological malignancies. Among the rapidly expanding list of PML-associated biologics, natalizumab (Tysabri®) has the highest incidence and is an ominous sequela for multiple sclerosis (MS) patients who otherwise benefit from dramatic reductions in relapses using this immunomodulatory agent. Drug withdrawal, the only therapeutic option for PML, is often complicated by a high-mortality cerebral inflammatory reaction. No anti-JCPyV agents are available. Polyomaviruses are species-specific. Lack of a tractable animal model of polyomavirus-induced CNS disease is an acknowledged bottleneck to elucidating PML pathogenesis, the immunological mechanisms that control JCPyV, in vivo evaluation of agents that inhibit polyomavirus replication in tissue culture, and uncovering early events that presage irreversible JCPyV-associated neuropathology. Using mouse polyomavirus (MuPyV), we developed a natural virus-host model of polyomavirus-associated CNS disease. In this R35 application, we plan to leverage our three recent key findings: (1) Mapping JCPyV-PML VP1 capsid protein mutations to MuPyV’s VP1 confers escape from virus- neutralizing antibodies (nAb) while preserving CNS tropism; (2) IL-21 produced by high-affinity anti-MuPyV CD4 T cells in the brain is required for formation and maintenance of MuPyV-specific brain resident-memory CD8 T cells (bTRM); and (3) STAT1-dependent innate immunity limits infection of the ventricular ependyma, a critical barrier to infection of the brain parenchyma. These findings lay the foundation for three key questions to be addressed here: (1) Is the ependyma the staging ground for polyomavirus invasion of the brain parenchyma?; (2) Does the integrity of the CD8 bTRM response to persistent infection depend on subset heterogeneity?; and (3) Does T cell deficiency open the door for outgrowth of nAb-escape virus variants? The proposed studies will make use of cutting edge advances in next-generation sequencing to uncover rare VP1 mutations in vivo, custom cryo EM image reconstruction approaches to define endogenous VP1 nAb epitopes and nAb escape mechanisms, and high-resolution 3D imaging of intact mouse brains to visualize virus CNS entry and spread. Findings from these studies will answer fundamental questions about innate and adaptive immune control of polyomavirus CNS infection and conditions underlying dissemination of virus from the periphery into the brain before development of irreversible neuropathology.

抽象的 JC多瘤病毒（JCPYV）是一种无处不在的人类病原体，引起了几种毁灭性的脑部疾病 免疫受损的个体。这些JCPYV相关的中枢神经系统疾病中最明显的是 PML，一个 艾滋病定义的病变在卡特前时期已成为患者的生命并发症 接受免疫调节剂用于自身免疫和炎症性疾病，并治疗某些 血液学恶性肿瘤。在迅速扩展的PML相关生物制剂清单中，Natalizumab （tysabri®）发病率最高，是多发性硬化症患者（MS）患者的续集 否则，使用这种免疫调节剂中的复发中的急剧减少受益。戒毒， PML的唯一热选择，通常会因高病态性脑炎症反应而变得复杂。不 可以使用抗jcpyv代理。多瘤病毒是特定于规格的。缺乏可拖动的动物模型 多组病毒诱导的中枢神经系统疾病是阐明PML发病机理的公认瓶颈 控制JCPYV的免疫机制，在体内评估抑制多瘤病毒的药物 在组织培养中的复制，并发现预示不可逆的JCPYV相关的早期事件 神经病理学。使用小鼠多聚病毒（MUPYV），我们开发了一种天然病毒宿主模型 多组病毒相关的中枢神经系统疾病。在此R35应用程序中，我们计划利用我们最近的三个密钥 调查结果：（1）映射JCPYV-PML VP1 CAPSID蛋白突变为Mupyv的VP1赋予从病毒中逃脱的VP1 中和抗体（NAB），同时保存中枢神经系统抗体； （2）由高亲和力抗Mupyv产生的IL-21 大脑中的CD4 T细胞是形成和维持MUPYV特异性脑居民居民记忆所必需的 CD8 T细胞（BTRM）； （3）Stat1依赖性的先天免疫学限制了心室室心感染，A 大脑实质感染的关键障碍。这些发现为三个关键问题奠定了基础 在此处解决：（1）是decendyma，是脑部多瘤病毒侵入的阶段地面 实质？ （2）CD8 BTRM对持续感染的响应的完整性取决于子集 异质性？ （3）T细胞缺乏是否为Nab-escape病毒变体的生长打开了大门？这 拟议的研究将利用下一代测序中的前沿进步来发现罕见的VP1 体内突变，自定义的冷冻图像重建方法定义内源VP1 NAB表位 和NAB逃生机制，以及完整小鼠大脑的高分辨率3D成像，以可视化病毒CNS 进入并传播。这些研究的发现将回答有关先天和适应性的基本问题 多瘤病毒中枢神经系统感染的免疫控制以及病毒传播的条件从 在发展不可逆的神经病理学之前进入大脑的周围。