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: 10433973
• 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/10433973
• 关键词: 2019-nCoV; ACE2; Affect; Anosmia; Antibodies; Ataxia; Attenuated; Autopsy; Biology; Blood; Blood Circulation; Brain; Brain Stem; COVID-19; COVID-19 diagnosis; COVID-19 impact; COVID-19 pandemic; COVID-19 patient; COVID-19 treatment; Cardiopulmonary; Case Series; Cells; Central Nervous System Infections; Cerebrospinal Fluid; China; Communities; Coronavirus; Data; 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 infections; Maps; Measurement; Mediating; Modeling; Monitor; Mus; Nervous System Trauma; Nervous system structure; Neuraxis; Neurologic; Neurologic Effect; Neurologic Symptoms; Neurological Models; Neurons; Neurotropism; Olfactory dysfunction; 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 Disease; Respiratory Failure; Respiratory Tract Infections; Role; SARS coronavirus; SARS-CoV-2 infection; 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; Virus; Wild Type Mouse; adeno-associated viral vector; antigen detection; associated symptom; biobank; brain tissue; central nervous system injury; common symptom; coronavirus disease; cytokine; detection assay; human tissue; immunoreaction; influenzavirus; insight; interest; monocyte; mortality; mouse model; nervous system disorder; neuroinflammation; neuron loss; neuropathology; neurotropic; novel; overexpression; patient subsets; pulmonary symptom; receptor; respiratory; response; single-cell RNA sequencing; success; viral detection

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 共同199的大流行对世界的人口产生了巨大的影响，并对 全球社区的经济，健康和心理。尽管它与SARS-COV-1有相似之处，但 由SARS-COV-2引起的病理生理学的程度尚不清楚。特别是肺外表现 SARS-COV-2感染的影响仍然很少了解。来自中国和欧洲的案例系列表明 中枢神经系统与至少一部分患者参与了疾病过程，一些报告 估计多达30％的CoVID-19患者患有神经系统症状，包括癫痫发作，棘手 头痛，气味和味道受损。尽管有报道说Covid-19中有神经系统疾病 患者，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性和神经炎症 与该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 可能通过侵袭中枢神经系统或增加炎症引起神经系统症状 中枢神经系统内的细胞因子。我们假设SARS-COV-2感染具有神经侵染潜力和铅 在感染个体的中枢神经系统中改变和高炎性免疫抑制物。我们进一步假设 中枢神经系统的感染通过表达ACE2的直接毒性加剧了呼吸功能障碍 神经元是心肺功能的关键调节剂。我们的投资将结合 人类研究使用小鼠模型的人进行研究，我们可以轻松地管理病毒和评估 病理生理学。 AIM 1，我们将确定COVID-19患者的CNS免疫反应 结合单细胞RNA测序，细胞因子分析，病毒 测序和抗体验证，我们将完全剖析中枢神经系统内的炎症反应 与Covid-19患者的全身循环相比，隔室。在AIM 2中使用鼠标模型，我们 将研究SARS-COV-2的脑潜力。使用几种完整的方法 用SARS-COV-2感染小鼠，我们将将病毒引入小鼠中枢神经系统。使用 耗尽抗体和各种敲除小鼠，我们将确定需要哪些免疫细胞 通过生存研究，流式细胞术和免疫荧光染色，这些小鼠的神经病理学。最后， 在AIM 3中，我们将评估中枢神经系统感染对呼吸结局的影响。因为已知 ACE2在脑干中的表达以及脑干在控制心肺功能中的关键作用， 我们怀疑使用SARS-COV-2感染的CNS感染会加剧SARS-COV-2呼吸道疾病。 这三个目标将有助于支持我们的假设，即SARS-COV-2感染如何影响中枢神经系统和 呼吸隔室。我们希望我们的发现将发现新的策略来治疗诊断的患者 借助Covid-19，并有助于获得新的见解，以了解SARS-COV-2病理生理学的生物学。