Human coronavirus infection of the nasal epithelium

鼻上皮的人冠状病毒感染

基本信息

批准号：
10708154
负责人：
Noam A Cohen
金额：
$ 74.35万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-20 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708154
关键词：
2019-nCoV Address Affect Air Amino Acid Sequence Antiviral Response Biological Biology COVID-19 COVID-19 pandemic COVID-19 risk Cell Line Cell membrane Cells China Clinical Complement Coronavirus Coronavirus Infections Cytopathology Cytoplasm Data Disease Disease Outbreaks Endosomes Epithelial Cells Epithelium Functional disorder Future Genes Genetic Polymorphism Genetic Transcription Genome Genotype Human Immune response Individual Infection Kinetics Leucine Zippers Link Liquid substance Lung Lung infections Middle East Respiratory Syndrome Middle East Respiratory Syndrome Coronavirus Modeling Nasal Epithelium Nitric Oxide Nose Outcome Pathogenicity Pathway interactions Pattern Phenotype Pneumonia Positioning Attribute Production Productivity Prophylactic treatment Protein Isoforms Proteins Reporting Resistance Resources Ribonucleases Risk Role Route SARS coronavirus SARS-CoV-2 B.1.1.529 SARS-CoV-2 infection Scheme Severities Severity of illness Site Structure TMPRSS2 gene Temperature Testing Therapeutic Upper Respiratory Infections Upper respiratory tract Variant Viral Virus Virus Replication Virus Shedding Work adaptive immune response airway epithelium alveolar type II cell antimicrobial peptide biobank cell type cold temperature cytokine design disorder risk experience genome wide association study genomic locus human coronavirus novel prevent respiratory response risk variant sensor single-cell RNA sequencing transcription factor viral RNA

项目摘要

Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 emerged in China in late 2019, resulting in the COVID-19 pandemic. Like SARS-CoV (2002) and Middle East respiratory syndrome (MERS)-CoV (2012), SARS-CoV-2 can progress to cause lethal pneumonia. In contrast, infections with “common” respiratory CoVs (NL63, 229E, OC43) are largely limited to the upper respiratory tract. Furthermore, SARS-CoV-2 and in particular the omicron variant, can sometimes cause primarily upper respiratory infections. Thus, despite their highly conserved genome structure and shared replication schemes, human CoVs induce varying degrees of disease. Respiratory CoVs initiate infection through the nose, though few studies have addressed CoV infection of the nasal epithelium. We have an established cryobank of nasal epithelial cells from over 1000 genetically characterized individuals capable of being expanded and grown as air liquid interface (ALI) cultures, recapitulating the nasal respiratory epithelium. Our preliminary studies demonstrate that SARS-2 (and its emerging variants), MERS and NL63 all productively infect these cultures. However, NL63 only replicates at a lower temperature (33C), infects single cells rather than clusters (evinced by SARS-2/MERS) and causes a more cytopathic effect than SARS-2 or MERS, suggesting it may induce a robust local immune response thereby limiting its replication to the upper respiratory tract or stimulating an adaptive immune response prior to infecting the lower airway. One COVID-19 risk locus includes the leucine zipper transcription factor-like 1 gene (LZTFL1), which we show is highly expressed in ciliated nasal cells, with ubiquitous expression throughout the cytoplasm. Our preliminary data of SARS-CoV-2 infected cultures genotyped for the high vs low-risk LZTFL1 polymorphisms demonstrate that LZTFL1 could play a role in variability of SARS-CoV-2 spread. In addition, polymorphisms in OAS1, a sensor of double-stranded viral RNA that initiates the antiviral RNase L pathway, have been linked to COVID-19 resistance. We have extensive experience in this pathway and recently reported that SARS-CoV-2 activates RNase L while MERS-CoV shuts it down. Based on these and other data, we hypothesize that pathogenic outcomes of CoV infections are reflected in viral biology in the nasal epithelium. Thus, using a battery of diverse CoVs we will assess differences in cell entry and spread, optimal temperature for viral replication and shedding as well as host nasal cell responses to each CoV. We propose to use our biobank to identify host and viral factors affecting the establishment of infection, host cytokine and nasal antiviral responses and the contribution of polymorphisms in LZTFL1 and OAS1 genes in the outcome of infection. Our complementary expertise in coronavirus biology (Weiss) and nasal pathophysiology (Cohen) uniquely positions us to address these Aims. This work will contribute to understanding nasal CoV infection, the divergence of lethal and common CoVs as well as variation in clinical course among SARS-CoV-2 infections, and may lead to novel targeted prophylaxis or therapeutic strategies targeting the nose, the site of initial contact.

严重的急性呼吸道综合征冠状病毒（SARS-COV）-2在2019年底出现在中国，导致 2019冠状病毒病大流行。像SARS-COV（2002）和中东呼吸道综合症（MERS）-COV（2012）一样， SARS-COV-2可以进展引起致命性肺炎。相反，具有“常见”呼吸道COV的感染（NL63，229E，OC43）在很大程度上仅限于上呼吸道。此外，SARS-COV-2，特别是 Omicron变体有时会引起原发性上呼吸道感染。那，他们很高保守的基因组结构和共享复制方案，人类COV诱导不同程度的疾病。呼吸道COV通过鼻子引发感染，尽管很少有研究解决了COV的感染鼻上皮。我们有一个从1000多个超过1000多个的鼻上皮细胞建立的冷冻库能够作为空气液体界面（ALI）培养的人的特征，能够扩展和生长，概括鼻呼吸道上皮。我们的初步研究表明SARS-2（及其新兴变体），MER和NL63都有生效的感染这些培养物。但是，NL63仅在较低的温度（33C），感染的单细胞而不是簇（由SARS-2/MER表现出来），并引起更多比SARS-2或MERS的细胞疗法作用，表明它可能引起稳健的局部免疫反应将其复制限制为上呼吸道或在感染之前刺激自适应免疫响应下气道。一个COVID-19风险基因座包括亮氨酸拉链转录因子样1基因（LZTFL1），我们显示的是在纤毛鼻细胞中高度表达的，在整个细胞质中具有无处不在的表达。我们针对高风险LZTFL1多态性的SARS-COV-2感染培养物的初步数据证明LZTFL1可以在SARS-COV-2传播的变异性中发挥作用。另外，多态性 OAS1是启动抗病毒RNase L途径的双链病毒RNA的传感器，已与 COVID-19阻力。我们在这一途径上有丰富的经验，最近报道说SARS-COV-2 激活RNase L，而MERS-COV将其关闭。基于这些和其他数据，我们假设 COV感染的致病结果反映在鼻上皮的病毒生物学中。那，使用我们将评估一个潜水员COV的电池，我们将评估细胞进入和传播的差异，病毒的最佳温度复制和脱落以及宿主鼻细胞对每个COV的反应。我们建议使用我们的生物库识别影响感染建立的宿主和病毒因素，宿主细胞因子和鼻抗病毒 LZTFL1和OAS1基因在感染结果中的反应和多态性的贡献。我们的冠状病毒生物学（WEIS）和鼻病理生理学（Cohen）的互补专业知识独特的位置我们要解决这些目标。这项工作将有助于理解鼻COV感染，致命的差异 SARS-COV-2感染之间的常见COV以及临床过程的变化，并可能导致新颖针对鼻子的靶向预防或治疗策略，即初始接触部位。