COVID-19 and Kidney Injury: Urinary Transcriptomics of Kidney Injury in Novel Nonhuman Primate Models of SARS-CoV-2

COVID-19 和肾损伤：SARS-CoV-2 新型非人灵长类动物模型中肾损伤的尿转录组学

• 批准号: 10689671
• 负责人: Minolfa C Prieto
• 金额: $ 19.79万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-24 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10689671
• 关键词: 2019-nCoV; Acute; Acute Renal Failure with Renal Papillary Necrosis; Affect; African; Anatomy; Angiotensins; Animal Model; Biological Markers; CCL2 gene; COVID-19; COVID-19 impact; COVID-19 patient; COVID-19 susceptibility; Cell Separation; Cell physiology; Cells; Cessation of life; Chad; Chronic Kidney Failure; Clinical; Clinical Markers; Communicable Diseases; Complication; Coronavirus; Creatinine; Critical Illness; Detection; Deterioration; Development; Disease Progression; Disease model; Doctor of Philosophy; Duct (organ) structure; Early Diagnosis; Early Intervention; Epidermal Growth Factor; Epithelium; Etiology; Excretory function; Family; Gelatinase A; Gene Expression; Gene Expression Profile; Glomerular capsule structure; Growth; Growth Factor; Hospital Mortality; Hospitalization; Human; IL18 gene; Immune response; Impairment; Infection; Injury to Kidney; Kidney; Kidney Diseases; Lung; Macaca mulatta; Measurement; Mechanical ventilation; Modeling; Nose; Organ; Outcome; Parietal; Patients; Persons; Phase; Plasminogen Activator; Play; Prevalence; Primates; Prognosis; Proteinuria; Public Health; Recovery; Renal function; Renin-Angiotensin System; Research; Research Personnel; Resolution; Role; SARS coronavirus; SARS-CoV-2 infection; Sampling; Serum; Signal Transduction; Structure; System; Testing; Therapeutic; Transforming Growth Factors; Tubular formation; Urine; Urokinase Plasminogen Activator Receptor; Viral Respiratory Tract Infection; Virus; Work; cardiovascular effects; cell type; current pandemic; cytokine release syndrome; glomerular function; member; mesangial cell; mortality; nephrotoxicity; next generation; nonhuman primate; novel; organ injury; pandemic disease; podocalyxin; podocyte; post gamma-globulins; previous outbreak; rat KIM-1 protein; receptor; severe COVID-19; severe injury; single-cell RNA sequencing; success; systemic inflammatory response; tool; transcriptome sequencing; transcriptomics; urinary; vaccine evaluation

ABSTRACT/SUMMARY The severe acute respiratory syndrome coronavirus-2 (SARS CoV-2), etiologic agent of the COVID-19, is the cause of the current worldwide pandemic with unprecedented public health issues. The lung is the most severely injured organ by SARS-CoV-2, but the kidney appears to be damaged as well. In 90% of patients with COVID-19 on mechanical ventilation, acute kidney injury (AKI) occurs early, leading to poor prognosis and high in-hospital mortality. Angiotensin (Ang) converting ezyme-2 (ACE2), a member of the renin-angiotensin system, opposes Ang II actions and exerts beneficial CV effects. There is mounting evidence that ACE-2, acts as a major entry receptor for SARS-CoV-2. Because ACE2 is expressed in proximal tubules, podocytes, mesangial cells, parietal epithelium of Bowman’s capsule and collecting ducts, it is critical to elucidate the effects of COVID-19 in the development of AKI. Animal models have been used extensively during previous outbreaks of SARS- CoV to model disease progression and to test vaccines and therapeutics. Non-human primates (NHPs) are ideally suited to model respiratory viral infections in humans primarily due to their similarities in anatomy and immunologic responses. Chad J. Roy, PhD (Director of Infectious Disease Aerobiology, Tulane National Primate Research Center, and Co-Investigator of this project) has recently generated novel and translational NHP models of COVID-19 using SARS-CoV-2 infection via nasal/intratracheal inoculation, which is able to reproduce the rapid clinical deterioration seen in people with severe COVID-19. In this pilot exploratory study, we propose to assess urine transcriptomics in two novel NHP (Rhesus macaque and African green) models of SARS-2-CoV-2 infection and their associations with early markers of AKI. Our overall hypothesis is that, because ACE2 may play a critical role in COVID-19-induced kidney injury, changes in urinary transcriptional signatures are associated with alterations of urinary early kidney injury markers and reflect the effects of SARS-CoV-2 on kidney ACE2-expressing cells and renal function. Using urine transcriptomics assessed by single-cell RNA-sequencing in ACE2-expressing cells isolated from urines collected during early infection, progression, and recovery phases of COVID-19, urinary measurements of kidney injury markers (EGF, MCP-1, NGAL), and clinical parameters of kidney function, we propose the following Specific aim: To test the hypothesis that the effects of SARS-CoV2 on the kidney are evidenced by changes of urine transcriptomics and accompanied by early excretion of kidney injury markers in NHPs with SARS-CoV-2 infection by mean of: A) To assess the temporal changes of urinary kidney injury markers during early infection, progression, and recovery phases of COVID-19. B) To determine whether urinary excretion of early kidney injury markers precede clinical parameters of kidney impairment and AKI; and C) To examine changes of urine transcriptomics in ACE2-expressing cells at a single-cell resolution during COVID-19. The success of the proposed study will help to elucidate the value of gene expression–based cell type enrichment analyses in urine as initial signals of renal injury during the progression of COVID-19.

摘要/总结 严重急性呼吸道综合征冠状病毒2型（SARS CoV-2）是COVID-19的病原体， 这是造成当前全球大流行的原因，带来了前所未有的公共卫生问题。肺是最 严重受损的器官，但肾脏似乎也受到损害。90%的患者 COVID-19对机械通气，急性肾损伤（阿基）发生早，导致预后差，高 住院死亡率。 血管紧张素转换酶2（ACE 2）是肾素-血管紧张素系统的一员， 并发挥有益的CV效应。越来越多的证据表明，ACE-2，作为一个主要的入口， SARS-CoV-2的受体。由于ACE 2在近端小管、足细胞、系膜细胞中表达， Bowman囊和集合管的壁上皮，这对于阐明COVID-19的影响至关重要 阿基的发展。在以往的SARS爆发期间，动物模型被广泛使用- CoV用于模拟疾病进展并测试疫苗和治疗方法。非人类灵长类动物（NHP） 理想地适合于模拟人类呼吸道病毒感染，主要是由于它们在解剖学上的相似性， 免疫反应。Chad J. Roy博士（杜兰国家大学传染病空气生物学主任） 灵长类动物研究中心和该项目的共同研究者）最近产生了新的和翻译的 使用SARS-CoV-2通过鼻内/肠道接种感染的COVID-19 NHP模型，其能够 重现了严重COVID-19患者的快速临床恶化。在这项初步探索性研究中， 我们建议在两种新的NHP（恒河猴和非洲绿色）模型中评估尿液转录组学， SARS-2-CoV-2感染及其与阿基早期标志物的相关性我们的总体假设是， 由于ACE 2可能在COVID-19诱导的肾损伤中发挥关键作用，尿转录水平的变化 签名与尿早期肾损伤标志物的改变相关，并反映了 SARS-CoV-2对肾脏ACE 2表达细胞和肾功能的影响使用尿液转录组学评估， 从早期感染期间收集的尿中分离的表达ACE 2的细胞中的单细胞RNA测序， COVID-19的进展和恢复阶段，肾损伤标志物（EGF，MCP-1， NGAL）和肾功能的临床参数，我们提出以下具体目的： 假设SARS-CoV 2对肾脏的影响通过尿液转录组学的变化来证明 并伴有SARS-CoV-2感染的NHP中肾损伤标志物的早期排泄，其平均值为： A）评估在早期感染、进展和复发期间尿肾损伤标志物的时间变化。 COVID-19的恢复阶段。B）确定尿排泄的早期肾损伤标志物是否 在肾损伤和阿基的临床参数之前;和C）检查尿的变化 在COVID-19期间以单细胞分辨率在ACE 2表达细胞中进行转录组学研究。的成功 拟议的研究将有助于阐明基于基因表达的细胞类型富集分析的价值， 尿液是COVID-19进展期间肾损伤的初始信号。