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.

严重急性呼吸综合征冠状病毒 (SARS-CoV)-2 于 2019 年底在中国出现，导致像 SARS-CoV (2002) 和中东呼吸综合征 (MERS)-CoV (2012) 一样，COVID-19 大流行。相比之下，“常见”呼吸道冠状病毒感染可能会发展为致命性肺炎。（NL63、229E、OC43）主要局限于上呼吸道，尤其是 SARS-CoV-2。 omicron 变体有时可主要引起上呼吸道感染，尽管其高度。由于保守的基因组结构和共享的复制方案，人类冠状病毒会引起不同程度的疾病。呼吸道冠状病毒通过鼻子引发感染，尽管很少有研究涉及呼吸道冠状病毒感染我们拥有超过 1000 个鼻上皮细胞的冷冻库。表征能够作为气液界面（ALI）培养物进行扩展和生长的个体，我们的初步研究表明 SARS-2（及其）。（新兴变种）、MERS 和 NL63 都能有效感染这些培养物，但 NL63 只能以一定的速度复制。较低的温度（33°C），感染单个细胞而不是细胞群（SARS-2/MERS 证明）并导致更多细胞病变效应比 SARS-2 或 MERS 强，表明它可能由此诱导强大的局部免疫反应限制其复制到上呼吸道或在感染前刺激适应性免疫反应下呼吸道的一个 COVID-19 风险位点包括亮氨酸拉链转录因子样 1 基因 (LZTFL1)，我们发现它在纤毛鼻细胞中高度表达，并在整个细胞质中普遍表达。我们对 SARS-CoV-2 感染培养物进行高风险与低风险 LZTFL1 多态性基因分型的初步数据证明 LZTFL1 可能在 SARS-CoV-2 传播的变异性中发挥作用。 OAS1 是一种启动抗病毒 RNase L 途径的双链病毒 RNA 传感器，与我们在这一途径方面拥有丰富的经验，并且最近报道了 SARS-CoV-2。激活 RNase L，而 MERS-CoV 则将其关闭根据这些数据和其他数据，我们认为。冠状病毒感染的致病结果反映在鼻上皮的病毒生物学中。一组不同的冠状病毒，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种冠状病毒的反应我们建议使用我们的生物库。确定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药物 LZTFL1 和 OAS1 基因多态性对感染结果的反应和贡献。冠状病毒生物学（Weiss）和鼻病理生理学（Cohen）的互补专业知识独特的地位我们致力于解决这些目标，这项工作将有助于了解鼻腔冠状病毒感染、致死率的差异。和常见的 CoV 以及 SARS-CoV-2 感染之间临床病程的差异，可能会导致新的针对鼻子（最初接触的部位）的有针对性的预防或治疗策略。