Humoral Immune Mechanisms of Acute and Chronic Neurologic Sequelae of COVID-19

项目摘要 COVID-19与越来越多的外周和中枢神经系统并发症有关。 很明显，这些综合征的一个子集，包括急性坏死性脑病，类固醇- 反应性脑炎和格林-巴利综合征，可能是由于直接的病毒神经侵入和/或 由SARS-CoV-2引发的自身免疫迫切需要前瞻性地研究急性和 COVID-19的慢性神经系统并发症，并确定哪些综合征是神经炎性起源。 虽然SARS-CoV-2的抗病毒疗法仍在开发中，但自身免疫性CNS疾病可能非常严重。 对免疫抑制有反应。因此，鉴定COVID-19患者亚组的生物标志物， 自身免疫性CNS综合征尤其可能立即影响临床治疗。 在过去的8年里，一个独特的跨学科团队的神经学家和基础科学家在加州大学旧金山分校是 形成开发和部署一个综合的方法，以快速识别微生物核酸，抗病毒， 与脑炎相关的抗体和抗神经抗体，明确的目的是发现和 验证临床上可操作的生物标志物，以及揭示疾病的基本机制 这些综合征的发病机制。这些努力的核心是正在进行的患者队列， NID（神经炎性疾病）队列，由疑似感染性或炎性 脑炎该队列现在有超过1，400名患者由UCSF的临床医生和周围的其他中心转介 世界这个队列已经刺激了有史以来第一个临床验证的脑脊髓 流体宏基因组下一代测序分析，鉴定一种新的副肿瘤性自身免疫 一种对男性乳腺癌患者有重要意义的综合征和肠道病毒CSF的鉴定 急性弛缓性肌无力患儿的抗体水平。在这里，我们建议调整现有的临床研究， 和实验室基础设施，以招募和调查COVID-19患者是否 患有中枢神经系统炎症。我们将合作完成这项工作 与美国国立卫生研究院、耶鲁大学以及加州大学旧金山分校医学中心的同事们一起， 弗朗西斯科总医院和UCSF贝尼奥夫儿童医院。利用我们独特的临床和分子 我们将通过以下具体目标调查这一假设： 目的1：表征急性和慢性神经系统疾病的COVID-19患者CSF中的自身抗体 综合征 目的2：使用以下方法确定COVID-19患者中的CSF特异性抗体库 高分辨率SARS-CoV-2全蛋白组抗体谱分析 目的3：通过从克隆中产生单克隆抗体来阐明自身抗体的致病性 在COVID-19患者中扩增CSF B细胞，以开发疾病的动物模型

PROJECT SUMMARY COVID-19 is associated with a growing number of peripheral and central nervous system complications. It has become clear that a subset of these syndromes, including acute necrotizing encephalopathy, steroid- responsive encephalitis and Guillain-Barré syndrome, are likely due to direct viral neuroinvasion and/or autoimmunity triggered by SARS-CoV-2. There is an urgent need to prospectively investigate the acute and chronic neurologic complications of COVID-19 and determine which syndromes are neuroinflammatory in origin. While anti-viral therapeutics are still being developed for SARS-CoV-2, autoimmune CNS conditions can be very responsive to immunosuppression. Thus, identifying biomarkers for a subset of COVID-19 patients with autoimmune CNS syndromes in particular could immediately impact clinical management. Over the past 8 years, a unique interdisciplinary team of neurologists and basic scientists at UCSF was formed to develop and deploy an integrated approach to rapidly identify microbial nucleic acid, anti-viral antibodies and anti-neural antibodies associated with encephalitis, with the explicit intent to discover and validate clinically actionable biomarkers in addition to uncovering the fundamental mechanisms of disease pathogenesis underlying these syndromes. The centerpiece of these efforts is an ongoing patient cohort called the NID (Neuroinflammatory Disease) cohort, consisting of patients with suspected infectious or inflammatory encephalitis. This cohort is now >1,400 patients referred by clinicians at UCSF and from other centers around the world. Already, this cohort has spurred the development of the first ever clinically validated cerebrospinal fluid metagenomic next-generation sequencing assay, the identification of a novel paraneoplastic autoimmune syndrome with important implications for men with seminoma and the identification of enteroviral CSF antibodies in children with acute flaccid myelitis. Here, we propose to adapt this existing clinical research and laboratory infrastructure to enroll and investigate the urgent question whether COVID-19 patients with ongoing neurologic sequelae have CNS inflammation. We will perform this work in collaboration with colleagues at the NIH, Yale University as well as at UCSF Medical Center, Zuckerberg San Francisco General Hospital and UCSF Benioff Children’s Hospital. Using our unique clinical and molecular approach, we will investigate this hypothesis through the following specific aims: Aim 1: Characterize autoantibodies in the CSF of COVID-19 patients with acute and chronic neurologic syndromes Aim 2: Identify CSF specific antibody repertoires in COVID-19 patients with neurologic complications using high-resolution SARS-CoV-2 proteome-wide antibody profiling Aim 3: Elucidate autoantibody pathogenicity through production of monoclonal antibodies from clonally expanded CSF B cells in COVID-19 patients to enable the development of animal models of disease