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; 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/摘要 COVID-19的严重急性呼吸综合征冠状病毒-2（SARS COV-2）是 当前全球大流行的原因，存在前所未有的公共卫生问题。肺最多 SARS-COV-2严重受伤的器官，但肾脏似乎也受到损害。 90％的患者 COVID-19关于机械通气，急性肾脏损伤（AKI）的早期发生，导致预后不良和高度 住院死亡率。 血管紧张素（ANG）转化Ezyme-2（ACE2）是肾素 - 血管紧张素系统的成员，反对ANG II 行动并发挥有益的简历效应。有越来越多的证据表明ACE-2充当主要入境 SARS-COV-2的受体。由于ACE2在近端管，足细胞，肾小球细胞中表达，所以 鲍曼胶囊和收集管道的顶叶上皮，阐明Covid-19的影响至关重要 在Aki的发展中。在先前的SARS爆发期间，动物模型已被广泛使用 COV来建模疾病进展并测试疫苗和治疗。非人类灵长类动物（NHP）是 理想情况下，由于其在解剖学和 免疫反应。乍得·罗伊（Chad J. Roy） 灵长类动物研究中心以及该项目的共同研究者）最近产生了新颖和翻译 通过鼻内/运动内接种，使用SARS-COV-2感染的COVID-19的NHP模型，能够 繁殖在严重的Covid-19患者中发现的快速临床确定。在这项试点探索性研究中， 我们建议评估两个新型NHP（Rhesus Macaque和African Green）模型的尿液转录组学 SARS-2-COV-2感染及其与AKI早期标记的关联。我们的总体假设是， 因为ACE2可能在COVID-19诱导的肾脏损伤中起关键作用，所以尿转录的变化 签名与尿早期肾脏损伤标记的改变有关，并反映 SARS-COV-2在表达肾脏ACE2的细胞和肾功能上。使用由 从早期感染期间收集的尿液中分离出的ACE2表达细胞中的单细胞RNA序列， COVID-19的进展和恢复阶段，肾损伤标记的尿测量值（EGF，MCP-1， Ngal）和肾功能的临床参数，我们提出以下特定目的：测试 尿液转录组学的变化证明了SARS-COV2对肾脏的影响的假设 并通过在SARS-COV-2感染的NHP中早期排泄肾脏损伤标记以： a）评估早期感染，进展和 COVID-19的恢复阶段。 b）确定早期肾脏损伤标记的尿极端是否极端 先于肾脏损伤和AKI的临床参数； c）检查尿液的变化 在COVID-19期间，在单细胞分辨率的ACE2表达细胞中的转录组学。成功的成功 拟议的研究将有助于阐明基因表达的基于基因的细胞类型富集分析的价值 尿液作为共vid-19期间肾脏损伤的初始信号。