Viral and immune-mediated CNS pathology during SARS-CoV-2 infection

SARS-CoV-2 感染期间病毒和免疫介导的中枢神经系统病理学

• 批准号: 10160327
• 负责人: Shelli Farhadian
• 金额: $ 41.78万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-24 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10160327
• 关键词: 2019-nCoV; Affect; Antibodies; Ataxia; Attenuated; Autopsy; Biological Assay; Biology; Blood; Blood Circulation; Brain; Brain Stem; COVID-19; COVID-19 pandemic; Cardiopulmonary; Case Series; Cells; Central Nervous System Infections; Cerebrospinal Fluid; China; Communities; Coronavirus; Data; Detection; Diagnosis; Disease; Dose; Encephalopathies; Enrollment; Europe; Evaluation; Family member; Fever; Flow Cytometry; Functional disorder; Genomics; Goals; Headache; Health; Heart; Heart Hypertrophy; Histology; Histopathology; Human; Hypertension; Hypogeusia; Hypoxemia; Immune; Immune response; Impairment; Individual; Infection; Inflammatory; Inflammatory Response; Institutional Review Boards; Intractable Headaches; Invaded; Investigation; Knockout Mice; Lead; Lung; Lung diseases; Lung infections; Maps; Measurement; Mediating; Modeling; Monitor; Mus; Nervous System Trauma; Nervous system structure; Neuraxis; Neurologic; Neurologic Effect; Neurologic Symptoms; Neurological Models; Neurons; Neurotropism; Outcome; Pathogenicity; Pathology; Pathway interactions; Patients; Physiological; Plaque Assay; Plasma Cells; Pneumonia; Population; Process; Production; Protocols documentation; Pulmonary Heart Disease; Pulmonary Pathology; Reporting; Respiration Disorders; Respiratory Failure; Respiratory Signs and Symptoms; Respiratory Tract Infections; Role; SARS coronavirus; Sampling; Secondary to; Seizures; Smell Perception; Spinal Puncture; Stains; Structure of parenchyma of lung; Study models; Supporting Cell; Symptoms; Taste Perception; Tissues; Toxic effect; Tumor-infiltrating immune cells; Validation; Viral; Viral Antigens; Virus; Wild Type Mouse; adeno-associated viral vector; associated symptom; biobank; brain tissue; central nervous system injury; common symptom; coronavirus disease; cytokine; human tissue; immunoreaction; influenzavirus; insight; interest; monocyte; mortality; mouse model; nervous system disorder; neuroinflammation; neuron loss; neuropathology; neurotropic; novel; overexpression; patient subsets; receptor; respiratory; response; single-cell RNA sequencing; success

Project Summary The COVID-19 pandemic has rampantly affected the population of the world and created lasting effects on the economy, health and psyche of the global community. Although it shares similarities with SARS-CoV-1, the full extent of the pathophysiology caused by SARS-CoV-2 is unclear. In particular, extrapulmonary manifestations effects of SARS-CoV-2 infection remain poorly understood. Case series from China and Europe suggest that the central nervous system is involved in the disease process in at least a subset of patients, with some reports estimating up to 30% of COVID-19 patients having neurological symptoms, including seizure, intractable headache, and impaired smell and taste. Although there are reports of neurological disease in in COVID-19 patients, it is unclear if SARS-CoV-2 invades the central nervous system (CNS). Studies of other coronaviruses, including SARS-CoV-1, demonstrate clear neurotropism as well as neuroinflammation associated with other members of this family of viruses. These studies raise the possibility that SARS-CoV-2 may cause neurological symptoms either through invasion of the CNS or through an increase in inflammatory cytokines within the CNS. We hypothesize that SARS-CoV-2 infections have neuroinvasive potential and lead to altered and hyperinflammatory immune states within the CNS of infected individuals. We further hypothesize that infection of the CNS exacerbates respiratory dysfunction through direct toxicity of ACE2 expressing neurons that are critical regulators of cardiopulmonary function. Our investigations will combine the power of human studies with those utilizing mouse models in which we can readily administer virus and assess for pathophysiology. Aim 1, we will determine the CNS immune responses in COVID-19 patients with neurological symptoms. Using a combination of single cell RNA-sequencing, cytokine profiling, viral sequencing and antibody validations, we will fully dissect out the inflammatory responses within the CNS compartment compared to the systemic circulation in COVID-19 patients. Using mouse models, in Aim 2, we will investigate the encephalitic potential of SARS-CoV-2. Using several complementary approaches to infect mice with SARS-CoV-2, we will introduce the virus into the central nervous system of mice. Using depletion antibodies and various knockout mice, we will identify which immune cells are required for neuropathology in these mice through survival studies, flow cytometry and immunofluorescent staining. Finally, in Aim 3, we will evaluate the effects of CNS infection on respiratory outcomes. Because of the known expression of ACE2 in the brainstem, and the brainstem’s critical role in regulating cardiopulmonary functions, we suspect that CNS infection with SARS-CoV-2 will exacerbate SARS-CoV-2 respiratory disease. These three aims will help support our hypotheses of how SARS-CoV-2 infections can affect the CNS and respiratory compartments. We expect that our findings will uncover new strategies to treat patients diagnosed with COVID-19 and help gain new insight to understanding the biology of SARS-CoV-2 pathophysiology.

项目摘要 COVID-19大流行严重影响了世界人口，并对全球经济产生了持久影响。 全球社会的经济、健康和心理。虽然它与SARS-CoV-1有相似之处，但完整的 SARS-CoV-2引起的病理生理学程度尚不清楚。特别是肺外表现 对SARS-CoV-2感染的影响仍然知之甚少。来自中国和欧洲的病例系列表明， 至少在一部分患者中，中枢神经系统参与疾病过程， 估计高达30%的COVID-19患者有神经系统症状，包括癫痫发作，顽固性 头痛，嗅觉和味觉受损。尽管有报告称，在COVID-19中， 目前尚不清楚SARS-CoV-2是否侵入中枢神经系统（CNS）。其他研究 包括SARS-CoV-1在内的冠状病毒表现出明显的神经嗜性和神经炎症 与这个病毒家族的其他成员有关。这些研究提出了SARS-CoV-2的可能性 可通过侵入CNS或通过炎性细胞因子增加引起神经系统症状。 CNS内的细胞因子。我们假设SARS-CoV-2感染具有神经侵袭潜力， 感染者中枢神经系统内免疫状态的改变和高度炎症。我们进一步假设 CNS感染通过表达ACE 2的直接毒性加重呼吸功能障碍， 神经元是心肺功能的关键调节器。我们的调查将结合联合收割机 人类研究与那些利用小鼠模型的研究，在小鼠模型中我们可以容易地施用病毒并评估 病理生理学目的1，我们将确定COVID-19患者的CNS免疫应答， 神经系统症状使用单细胞RNA测序、细胞因子谱分析、病毒 通过测序和抗体验证，我们将充分剖析CNS内的炎症反应 与COVID-19患者的体循环相比，使用小鼠模型，在目标2中，我们 将研究SARS-CoV-2的脑炎潜力。使用几种互补的方法， 用SARS-CoV-2感染小鼠，我们将把病毒导入小鼠的中枢神经系统。使用 耗尽抗体和各种敲除小鼠，我们将确定哪些免疫细胞是必需的， 通过存活研究、流式细胞术和免疫荧光染色来观察这些小鼠的神经病理学。最后， 在目标3中，我们将评估CNS感染对呼吸系统结局的影响。由于已知的 ACE 2在脑干中的表达，以及脑干在调节心肺功能中的关键作用， 我们怀疑SARS-CoV-2的CNS感染会加重SARS-CoV-2呼吸道疾病。 这三个目标将有助于支持我们关于SARS-CoV-2感染如何影响CNS的假设， 呼吸腔室我们希望我们的研究结果将揭示新的策略，以治疗诊断为癌症的患者。 并帮助获得新的见解，以了解SARS-CoV-2病理生理学的生物学。