How the Novel Coronavirus Attacks the Brain

新型冠状病毒如何攻击大脑

• 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
• 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' s disease pathology; Anosmia; Anterior; Astrocytes; Attention; Behavioral; Blood Vessels; Brain; Brain Injuries; Brain Pathology; Brain region; Brodmann' 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

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多种多样且独特 表现，包括嗅觉丧失，混乱，和改变精神状态，当小说的过程中， 冠状病毒病（COVID-19）因对神经系统的损害而变得复杂。鼻咽和 鼻腔是高病毒载量的储存库，嗅觉组织含有关键的受体和蛋白酶， 可以促进病毒进入细胞并在细胞水平复制。脑细胞的下游机制 对SARS-CoV-2入侵和整合仍然知之甚少，特别是SARS-CoV-2如何引起扩散 神经影响COVID-19患者持续严重的细胞因子风暴， 炎症和凝血结合内皮损伤，可能导致血栓栓塞事件， 小胶质细胞活化导致神经元损伤。患者还存在长期的脑后遗症， 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。 适当评估死亡和未死亡的COVID-19患者的特定大脑区域 存在神经系统症状将允许改善对可能的目标的理解，以限制大脑 损害此外，从SARS-CoV-2如何影响大脑的经验教训可以提供对可推广的 神经炎症对神经退行性疾病的作用机制。我们的目标是确定： 1.新冠肺炎患者在入院时有神经系统表现（NP-COVs）是否发生了改变 与没有COVs的人相比，调节炎症和凝血的基因的脑表达， 年龄和性别匹配的非COVID-19对照组（CONT）。我们将绘制整个转录组的图谱， 脑组织切片，使用单核RNA测序（sn-RNA-seq，10 X Genomics）。我们将验证和 使用Duplex定量神经元、神经胶质和血管相关细胞上的候选mRNA表达 RNAscope®（ACDBio），正如我们在CONT成功进行的那样。2. NP-COV是否具有升高的大脑 促炎标记物。我们将运行人细胞因子/趋化因子/生长因子组（48 μ g试剂盒， Milliopre）并定量细胞因子、趋化因子和生长因子。我们将绘制它们在神经元上的表达 和胶质细胞，使用双重免疫组织化学（IHC），因为我们在CONT试点。3.如果NP-COV升高， 脑小胶质细胞激活。使用双重IHC检测小胶质细胞标志物TSPO（转运蛋白）、CD 11b、Iba 1 （离子化钙结合衔接分子），神经元标记物和细胞定量的体视学，我们 将计算激活（变形虫）和静息（小细胞体和精细薄过程）小胶质细胞， 将空间关系映射到神经元。4.如果NP-COV具有减少的神经元密度和树突树枝状化。 使用双重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。