Functional characterization of SARS-CoV-2 myeloid cell receptors as an immunopathogenic mechanisms of COVID-19

SARS-CoV-2 骨髓细胞受体作为 COVID-19 免疫致病机制的功能特征

基本信息

批准号：
10443833
负责人：
JUN WANG
金额：
$ 21.19万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
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：
2021-07-02 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10443833
关键词：
2019-nCoV ACE2 Acute Respiratory Distress Syndrome Address Affect Affinity B-Lymphocytes Binding C-Type Lectins CD209 gene COVID-19 COVID-19 pandemic COVID-19 pathogenesis COVID-19 patient COVID-19 prevention COVID-19 severity COVID-19 treatment Cell Communication Cell Line Cells Coculture Techniques Collaborations Critical Pathways Data Data Set Dendritic Cells Disease Ectopic Expression Epithelial Cells Epitopes Family member Human Hyperactivity Immune Immune response Immune system Immunity Individual Infection Inflammation Inflammatory Response Lead Lectin Ligands Ligation Lung Lymphopenia Mediating Meta-Analysis Molecular Profiling Myeloid Cells New York Pathogenesis Pathogenicity Pathologic Pathway interactions Pattern Phenotype Play Polysaccharides Population Proteins Public Health Publishing Receptor Cell Receptor Signaling Role SARS-CoV-2 spike protein System T-Lymphocyte Testing Therapeutic Therapeutic antibodies Tissues Universities Viral Viral Pathogenesis Virus Virus Diseases Virus Receptors adaptive immunity base chemokine cytokine cytokine release syndrome experience experimental study functional outcomes immunopathology immunoregulation improved knock-down macrophage medical schools monocyte nanobodies new therapeutic target novel novel virus overexpression pandemic disease programs receptor receptor binding receptor expression response single cell sequencing single-cell RNA sequencing targeted treatment transcriptome sequencing

项目摘要

Project Summary The rapid spread of SARS-CoV-2 and the resulting Coronavirus Disease 2019 (COVID-19) pose a global pandemic. SARS-CoV-2 and its family members share some puzzling, unique pathological features, most notably acute respiratory distress syndrome (ARDS), cytokine release syndrome (CRS), and lymphopenia, despite excessive myeloid cell–dominant inflammation, which has been correlated with COVID-19 severity. How the virus engages and dysregulates the immune system is currently unknown; although angiotensin-converting enzyme 2 (ACE2) is the canonical SARS-CoV-2 receptor, immune cells, particularly myeloid cells, express little, if any, levels of ACE2 despite evidence for direct viral engagement. Based on our extensive experience in the discovery and characterization of immune-modulatory receptor-ligand interactions, we sought to address how SARS-CoV-2 interacts with myeloid cells, which play essential roles in both virus innate sensing and the modulation of host immunity, by developing a myeloid cell receptor-focused ectopic expression screen. In ongoing experiments, we have identified several novel glycan-dependent host interaction partners for SARS-CoV-2 Spike (S) protein, including several C-type lectins and Tweety Family Member 2 (TTYH2). Pulmonary single-cell RNA sequencing (scRNA-seq) analysis in COVID-19 patients indicates a myeloid cell-dominant expression of these receptors as opposed to ACE2. In the preliminary studies, we have shown that these molecules interact mostly through regions outside of the ACE2 receptor-binding domain (RBD), suggesting that they may provide a novel function outside of virus entry. Although these receptors do not support the active replication of authentic SARS-CoV-2, the direct virus-myeloid cell engagement induces a robust pro-inflammatory response, which is blocked by receptor-decoy proteins and a picomolar-affinity anti- spike bispecific nanobody that also blocks virus infection through ACE2. Together, our findings provide the first evidence for direct immune modulation by SARS-CoV-2, potentially targeted for therapeutic benefit. Given these findings, we hypothesize that novel SARS-CoV-2 virus-receptor interactions in myeloid cells constitute a pathogenic pathway for COVID-19, serving as signaling receptors that directly drive myeloid cell dysregulation. These new direct virus-immune interactions may also have heretofore unexplored functions to affect other cells indirectly. These hypotheses will be addressed within the following Specific Aims: (1) to determine the functional contribution of each myeloid cell receptor in the induction of pro-inflammatory responses upon SARS-CoV-2 engagement, and (2) to determine the role of these interactions in the dysregulation of adaptive immunity and SARS-CoV-2 trans-infection through ACE2 receptor. Our study on novel virus myeloid cell receptors that govern aberrant immune responses would greatly contribute to our understanding of COVID- 19 pathogenesis, revealing new therapeutic targets against COVID-19 for the benefit of humanity at large.

项目概要 SARS-CoV-2 的快速传播以及由此产生的 2019 年冠状病毒病 (COVID-19) 构成了全球性的公共卫生危机 SARS-CoV-2 及其家族成员有一些令人费解的、独特的病理特征，其中大多数具有共同特征。值得注意的急性呼吸窘迫综合征（ARDS）、细胞因子释放综合征（CRS）和淋巴细胞减少症，尽管存在过度的骨髓细胞为主的炎症，这与 COVID-19 的严重程度相关。尽管血管紧张素转化，但该病毒是否会影响免疫系统并使其失调，目前尚不清楚；酶 2 (ACE2) 是典型的 SARS-CoV-2 受体，免疫细胞，特别是骨髓细胞表达很少，尽管有证据表明病毒直接参与，但 ACE2 水平（如果有）。基于我们在免疫调节受体-配体的发现和表征方面的丰富经验相互作用，我们试图解决 SARS-CoV-2 如何与骨髓细胞相互作用，骨髓细胞在通过开发以骨髓细胞受体为中心的病毒先天感应和宿主免疫调节在正在进行的实验中，我们已经鉴定了几种新型的聚糖依赖性宿主。 SARS-CoV-2 Spike (S) 蛋白的相互作用伙伴，包括几种 C 型凝集素和 Tweety Family 成员 2 (TTYH2)。COVID-19 患者的肺单细胞 RNA 测序 (scRNA-seq) 分析。表明这些受体在骨髓细胞中占主导地位，与 ACE2 不同。我们的区域显示这些分子主要通过 ACE2 受体结合外部相互作用域（RBD），表明它们可能在病毒进入之外提供新的功能。不支持真正的 SARS-CoV-2 的主动复制，病毒与骨髓细胞的直接接触会诱导强烈的促炎反应，可被受体诱饵蛋白和皮摩尔亲和力抗炎药阻断刺突双特异性纳米抗体也可以通过 ACE2 阻断病毒感染。 SARS-CoV-2 直接免疫调节的证据，可能具有治疗效果。鉴于这些发现，我们探究了骨髓细胞中新型 SARS-CoV-2 病毒与受体的相互作用构成了 COVID-19 的致病途径，作为直接驱动骨髓细胞的信号受体这些新的直接病毒免疫相互作用也可能具有迄今为止尚未探索的功能。这些假设将在以下具体目标中得到解决：（1）确定每种骨髓细胞受体在诱导促炎反应中的功能贡献 SARS-CoV-2 的参与，以及 (2) 确定这些相互作用在失调中的作用我们对新型病毒髓系的研究。控制异常免疫反应的细胞受体将极大地有助于我们对新冠病毒的理解 19 发病机制，揭示了针对 COVID-19 的新治疗靶点，造福全人类。