How the Novel Coronavirus Attacks the Brain

新型冠状病毒如何攻击大脑

基本信息

批准号：
10317754
负责人：
Maura Boldrini
金额：
$ 32.84万
依托单位：
NEW YORK STATE PSYCHIATRIC INSTITUTE DBA RESEARCH FOUNDATION FOR MENTAL HYGIENE, INC
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-14 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10317754
关键词：
2019-nCoV Acute Affect Age Alzheimer&apos s disease pathology Anosmia Anterior Astrocytes Attention Behavioral Blood Vessels Brain Brain Injuries Brain Pathology Brain region Brodmann&apos s area COVID-19 COVID-19 patient Calcium Binding Calcium ion Cell Nucleus Cells Clinical Coagulation Process Cognitive Comprehension Computerized Medical Record Confusion Coronavirus Data Dendrites Diffuse Dysautonomias Endothelial Cells Endothelium Evaluation Event Family Fatigue Fogs Follow-Up Studies Gene Expression Genes Genomics Growth Factor Headache Hippocampus (Brain)Hospitals Human ITGAM gene Immunohistochemistry Impaired cognition Impairment Infection Inflammation Inflammatory Injury Institution Intake Interleukin Suppression Interleukin-1 Interleukin-10 Interleukin-6 Investigation Lead Length Link Maps Memory Microglia Modeling Molecular Nasal cavity Nasopharynx Nerve Degeneration Neurodegenerative Disorders Neuroglia Neurologic Neurologic Effect Neurologic Symptoms Neurons Pathologic Patients Peptide Hydrolases Pericytes Pharmaceutical Preparations Phenotype Pilot Projects Prefrontal Cortex Process Proteins Recording of previous events Rest Role Running SARS-CoV-2 infection SARS-CoV-2 negative Serum Short-Term Memory Sleeplessness Smell Perception Societies Suicide Symptoms System TNF gene Testing Therapeutic Effect Thinness Time Tissues Viral Viral Load result base brain tissue cell injury chemokine cognitive function coronavirus disease cost cytokine cytokine release syndrome density differential expression executive function improved inflammatory marker insight mRNA Expression mental state neurofilament neuroinflammation neuropathology neuropsychiatric symptom neuropsychiatry novel coronavirus receptor sex spatial relationship transcriptome transcriptome sequencing

项目摘要

Presentations of patients infected with SARS-CoV-2 are varied and unique in their neurological manifestations, including loss of smell, confusion, and altered mental status, when the course of the novel coronavirus disease (COVID-19) is complicated by insults to the neurological system. The nasopharynx and nasal cavities are reservoirs for high viral load and olfactory tissue contains key receptors and proteases that may facilitate viral entry and replication at the cellular level. Downstream mechanisms of brain cellular invasion and integration remain poorly understood, particularly how SARS-CoV-2 may be instigating diffuse neurological effects. Patients with COVID-19 sustain a severe cytokine storm, the interplay between inflammation and coagulation combined with endothelial damage, may lead to thrombo-embolic events, and microglia activation leading to neuronal damage. Patients also present with long-term brain sequela of COVID-19, including “brain fog,” difficulties concentrating, impaired short-term and working memory, fatigue, headache, dysautonomia, and insomnia, and the neuropathological bases of these symptoms are unknown. Appropriate evaluation of specific brain regions from deceased patients with COVID-19 who did and did not present with neurological symptoms will allow for improved comprehension of possible targets to limit brain damage. Additionally, lessons from how SARS-CoV-2 affects the brain may provide insight into generalizable mechanisms for effects of neuroinflammation on neurodegenerative diseases. We aim to determine: 1. Whether COVID-19 patients with neurological presentations at the time of intake (NP-COVs) have altered brain expression of genes regulating inflammation and coagulation compared to those without (COVs) and non-COVID-19 age and sex matched controls (CONT). We will map the whole transcriptome in the entire brain tissue section using single nuclei RNA sequencing (sn-RNA-seq, 10X Genomics). We will validate and quantify candidate mRNAs expression on neurons, glia, and vasculature-associated cells, using Duplex RNAscope® (ACDBio), as we successfully performed in CONT. 2. Whether NP-COVs have elevated brain pro-inflammatory markers. We will run a Human Cytokine/Chemokine/Growth Factor Panel (48 Plex Kit, Milliopre) and quantify cytokines, chemokines and growth factors. We will map their expression on neurons and glia, using double immunohistochemistry (IHC), as we piloted in CONT. 3. If NP-COVs have elevated brain microglia activation. Using double-IHC for microglia markers TSPO (translocator protein), CD11b, Iba1 (Ionized calcium binding adaptor molecule), and neuronal markers, and stereology for cell quantification, we will compute activated (amoeboid) and resting (small cell body and elaborated thin processes) microglia, and map spatial relationship to neurons. 4. If NP-COVs have reduced neuronal density and dendrite arborization. Using double-IHC for neuronal marker NeuN and neurofilament, Stereoinvestigator and Neurolucida (MBF Inc.), will quantify neuron density, dendrite length and arborization, as in our pilot studies.

感染 SARS-CoV-2 的患者在神经系统方面的表现各不相同且独特当小说进展时，会出现一些表现，包括嗅觉丧失、意识混乱和精神状态改变冠状病毒病（COVID-19）因神经系统受到损害而变得复杂化。鼻腔是高病毒载量的储存库，嗅觉组织含有关键受体和蛋白酶，可能促进病毒在细胞水平的进入和复制。入侵和整合仍知之甚少，特别是 SARS-CoV-2 可能如何引发弥漫性感染 COVID-19 患者承受着严重的细胞因子风暴，两者之间的相互作用。炎症和凝血与内皮损伤相结合，可能导致血栓栓塞事件，并且小胶质细胞激活导致神经元损伤，患者还会出现长期脑部后遗症。 COVID-19，包括“脑雾”、注意力难以集中、短期记忆和工作记忆受损、疲劳、头痛、自主神经功能障碍和失眠，这些症状的神经病理学基础尚不清楚。对已故的 COVID-19 患者的特定大脑区域进行适当评估，这些患者有或没有出现神经系统症状将有助于提高对限制大脑的可能目标的理解此外，从 SARS-CoV-2 如何影响大脑中获得的经验教训可能有助于深入了解可推广的结论我们的目标是确定神经炎症对神经退行性疾病的影响机制： 1. COVID-19患者在摄入时的神经系统表现（NP-COV）是否发生了改变与没有（COV）的基因相比，调节炎症和凝血的基因的大脑表达我们将绘制整个转录组的非 COVID-19 年龄和性别匹配对照（续）。使用单核 RNA 测序（sn-RNA-seq、10X Genomics）进行脑组织切片我们将进行验证和验证。使用 Duplex 量化神经元、神经胶质细胞和脉管系统相关细胞上的候选 mRNA 表达 RNAscope® (ACDBio)，正如我们在 CONT 2 中成功执行的那样。NP-COV 是否具有升高大脑的作用。我们将运行人类细胞因子/趋化因子/生长因子面板（48 Plex 试剂盒， milliopre）并量化细胞因子、趋化因子和生长因子，我们将绘制它们在神经元上的表达图谱。和神经胶质细胞，使用双免疫组织化学 (IHC)，正如我们在 CONT 3 中试验的那样。如果 NP-COV 升高。使用双 IHC 检测脑小胶质细胞标记物 TSPO（易位蛋白）、CD11b、Iba1。（离子钙结合接头分子）、神经元标记物以及细胞定量的体视学，我们将计算激活的（变形虫）和静止的（小细胞体和精细的薄过程）小胶质细胞，以及 4. 绘制与神经元的空间关系。 NP-COV 是否降低了神经密度和树突树枝化。使用双 IHC 检测神经元标记物 NeuN 和神经丝、Stereoinvestigator 和 Neurolucida (MBF Inc.），将量化神经元密度、树突长度和树枝化，就像我们的试点研究一样。