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

项目摘要 急性肾损伤（阿基）是冠状病毒病19（COVID-19）的常见并发症，影响超过200人。 三分之一的COVID-19住院患者和高达90%的需要机械通气的患者。可能 COVID-19患者阿基的促成因素包括全身炎症和细胞因子释放、血流动力学 以及血管内凝血此外，一些报告表明存在SARS- 死于COVID-19的患者肾脏组织中的CoV-2病毒颗粒或病毒RNA，表明可能存在 肾脏局部病毒感染的作用。 已知SARS-CoV-2可以感染肾细胞系并在其中复制，初步数据显示， 可以从严重COVID-19和阿基患者的尿液中扩增，即使在病毒被清除后也是如此 在呼吸道中。对尿液中这些病毒RNA的遗传分析表明， SARS-CoV-2的弗林蛋白酶切割位点的缺失和突变，在超过180，000个 公共数据库中保存的呼吸道样本SARS-CoV-2基因组序列 GISAID。然而，据报道，这些突变发生在病毒在非洲绿色中传代后。 猴肾细胞系Vero-E6，提高了这些突变可能是积极选择的可能性， 病毒在肾细胞中复制。 该应用程序的主要目标是了解局部病毒感染和局部 通过利用我们在HIV相关肾脏研究方面的专业知识， 疾病我们的具体目标是：1）从尿样中分离SARS-CoV-2并对其进行遗传学鉴定 COVID-19患者的轻度、中度或重度疾病，2）探讨SARS-CoV- 2）肾细胞感染和尿炎症标志物;和3）确定基因突变对 病毒适应度 了解肾上皮细胞直接病毒感染的作用是设计干预措施的关键， 预防和治疗COVID-19中的阿基。从尿中分离的病毒突变体的进一步特征可以 提供了深入了解病毒的发病机制，并将告知抗病毒和预防治疗的设计， 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.