Human coronavirus infection of the nasal epithelium

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 新冠肺炎大流行。与SARS冠状病毒(2002)和中东呼吸综合征(MERS)冠状病毒(2012)一样， SARS-CoV-2病毒可以发展成致命性肺炎。相比之下，感染“普通”呼吸道冠状病毒的患者 (NL63、229E、OC43)主要限于上呼吸道。此外，SARS-CoV-2，特别是 奥米克龙的变种有时会引起主要的上呼吸道感染。因此，尽管他们的高度 由于保守的基因组结构和共享的复制方案，人类冠状病毒会导致不同程度的疾病。 呼吸道冠状病毒通过鼻部感染，尽管很少有研究涉及呼吸道冠状病毒感染 鼻腔上皮。我们已经建立了1000多个遗传鼻腔上皮细胞的冷藏库 以能够作为气液界面(ALI)培养物扩展和生长的个体为特征， 重述鼻腔呼吸道上皮。我们的初步研究表明，SARS-2(及其 新出现的变种)、MERS和NL63都能有效地感染这些文化。然而，NL63的复制速度仅为 较低的温度(33摄氏度)，感染单个细胞而不是集群(表现为SARS-2/MERS)，并导致更多 比SARS-2或MERS更具细胞病变效应，表明它可能因此诱导强大的局部免疫反应 将其复制限制在上呼吸道或在感染前刺激适应性免疫反应 下呼吸道。一个新冠肺炎风险基因座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腔细胞中高度表达，在整个细胞质中普遍表达。 SARS-CoV-2感染培养物高危和低危LZTFL1基因分型的初步数据 证明LZTFl1在SARS-CoV-2传播的可变性中发挥作用。此外，基因中的多态 OAS1是启动抗病毒核糖核酸酶L途径的双链病毒rna的传感器，已被连接到 新冠肺炎的阻力。我们在这一途径上有丰富的经验，最近报道了SARS-CoV-2 激活核糖核酸酶L，而MERS冠状病毒将其关闭。根据这些和其他数据，我们假设 冠状病毒感染的致病结果反映在鼻黏膜上皮的病毒生物学中。因此，使用 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个冠状病毒的反应。我们建议使用我们的生物库 确定影响宿主和病毒感染的因素，宿主细胞因子和鼻腔抗病毒药物 LZTFL1和OAS1基因多态性在感染结局中的作用和贡献我们的 在冠状病毒生物学(Weiss)和鼻部病理生理学(Cohen)方面的互补专业知识具有独特的地位 我们需要解决这些目标。这项工作将有助于理解鼻腔冠状病毒感染致死率的差异 和常见的冠状病毒以及SARS-CoV-2感染之间的临床病程差异，并可能导致新的 针对鼻部的针对性预防或治疗策略，也就是最初接触的部位。

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.