Understanding the interplay between local viral infection and local inflammation in COVID-19 kidney injury

项目摘要 急性肾损伤是冠状病毒病19型(新冠肺炎)的常见并发症，影响超过 三分之一的患者因新冠肺炎入院治疗，高达90%的患者需要机械通气。可能的 新冠肺炎急性心肌梗死的致病因素包括全身炎症、细胞因子释放、血流动力学 妥协和血管内凝血。此外，几份报告显示SARS的存在-- 新冠肺炎患者肾脏组织中的CoV-2病毒颗粒或病毒核糖核酸，表明 对肾脏局部病毒感染的作用。 SARS-CoV-2已知可以在肾脏细胞系中感染和复制，在初步数据中，我们发现病毒RNA 即使在病毒被清除后，严重的新冠肺炎和急性心肌炎患者的尿液中也可以扩增出这种病毒 在呼吸道里。对尿液中这些病毒RNA的遗传分析表明， SARS-CoV-2呋喃裂解位点的缺失和突变在18万多人中未被观察到 公共数据库中存储的呼吸道样本中的SARS-CoV-2基因组序列 格萨德。然而，据报道，这些突变是在病毒在非洲绿地传代后发生的。 猴肾细胞系Vero-E6，增加了以下这些突变可能被正面选择的可能性 病毒在肾脏细胞中复制。 这个应用程序的主要目标是了解局部病毒感染和局部病毒感染之间的相互作用。 利用我们在艾滋病相关肾脏研究中的专业知识在新冠肺炎相关肾损伤中的炎症反应 疾病。我们的具体目标是：1)从尿样中分离SARS-CoV-2并鉴定其基因特征 对新冠肺炎患者的轻、中、重度疾病进行分析，2)探讨SARS冠状病毒与病毒感染的关系。 2)肾细胞感染和尿炎性标志物；3)确定基因突变对 病毒式健身。 了解病毒直接感染肾上皮细胞的作用是设计干预措施的关键 预防和治疗新冠肺炎中的AKI。从尿液中分离出的病毒突变株的进一步鉴定可能 提供对病毒发病机制的洞察，并将为抗病毒和辅助治疗的设计提供信息 SARS-CoV-2感染。

PROJECT ABSTRACT Acute kidney injury (AKI) is a common complication of Coronavirus disease 19 (COVID-19), affecting more than a third of patients hospitalized with COVID-19 and up to 90% of those requiring mechanical ventilation. Possible contributors to AKI in patients with COVID-19 include systemic inflammation and cytokine release, hemodynamic compromise, and intravascular coagulation. Additionally, several reports have suggested the presence of SARS- CoV-2 viral particles or viral RNA in the kidney tissue of patients who died from COVID-19, suggesting a potential role for local viral infection in the kidney. SARS-CoV-2 is known to infect and replicate in kidney cell lines, and in preliminary data we show that viral RNA can be amplified from the urine of patients with severe COVID-19 and AKI, even after the virus has been cleared in the respiratory tract. Genetic analysis of those viral RNA in urine demonstrated a predominant pattern of deletions and mutations at the furin-cleavage site of SARS-CoV-2 that have not been observed in over 180,000 SARS-CoV-2 genome sequences from respiratory tract samples deposited in the publicly available database GISAID. However, those mutations have been reported to occur after virus passaging in the African green monkey kidney cell line Vero-E6, raising the possibility that those mutations might be positively selected following virus replication in kidney cells. The primary goal of this application is to understand the interplay between local viral infection and local inflammation in COVID-19-related kidney injury by leveraging our expertise in the study of HIV-related kidney disease. Our specific objectives are 1) to isolate and genetically characterize SARS-CoV-2 from urine samples of COVID-19 patients with mild, moderate or severe disease, 2) to explore the relationship between SARS-CoV- 2 infection of renal cells and urine inflammatory markers; and 3) to determine the impact of genetic mutations on viral fitness. Understanding the role of direct viral infection of renal epithelial cells is key to the design of interventions to prevent and treat AKI in COVID-19. Further characterization of the viral mutants isolated from urine may provide insight into viral pathogenesis and would inform the design of antiviral and adjunctive therapies for SARS-CoV-2 infection.