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: 10453220
• 负责人: Minolfa C Prieto
• 金额: $ 23.53万
• 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/10453220
• 关键词: 2019-nCoV; Acute; Acute Renal Failure with Renal Papillary Necrosis; Affect; African; Anatomy; Angiotensin II; Angiotensins; Animal Model; Biological Markers; CCL2 gene; COVID-19; Cell physiology; Cells; Cessation of life; Chad; Chronic Kidney Failure; Clinical; Clinical Markers; Communicable Diseases; Complication; Coronavirus; Creatinine; Critical Illness; Detection; Deterioration; Development; Disease Outbreaks; Disease Progression; Disease model; Doctor of Philosophy; Duct (organ) structure; Early Intervention; Epidermal Growth Factor; Epithelial; Etiology; Excretory function; Family; Gelatinase A; Gene Expression; Gene Expression Profile; Glomerular capsule structure; Growth; Growth Factor; Hospital Mortality; Human; Immune response; Impairment; Infection; Injury to Kidney; Interleukin-18; Kidney; Kidney Diseases; Lung; Macaca mulatta; Measurement; Mechanical ventilation; Modeling; Nose; 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; Testing; Therapeutic; Transforming Growth Factors; Tubular formation; Urine; Urokinase Plasminogen Activator Receptor; Viral Respiratory Tract Infection; Virus; Work; base; body system; cardiovascular effects; cell type; cytokine release syndrome; glomerular function; member; mesangial cell; mortality; nephrotoxicity; next generation; nonhuman primate; novel; organ injury; pandemic disease; podocalyxin; podocyte; post gamma-globulins; rat KIM-1 protein; receptor; 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 严重急性呼吸综合征冠状病毒-2 (SARS CoV-2) 是 COVID-19 的病原体， 当前全球大流行带来前所未有的公共卫生问题的原因是肺部。 SARS-CoV-2 严重损害了器官，但 90% 的患者的肾脏似乎也受到了损害。 COVID-19对机械通气、急性肾损伤（AKI）发生较早，导致预后不良且高 院内死亡率。 血管紧张素 (Ang) 转换酶-2 (ACE2) 是肾素-血管紧张素系统的成员，可对抗 Ang II 作用并发挥有益的 CV 作用 越来越多的证据表明 ACE-2 是一个主要的入口。 因为 ACE2 在近端肾小管、足细胞、系膜细胞中表达， 鲍曼氏囊和集合管的壁上皮，阐明 COVID-19 的影响至关重要 动物模型的开发主要在之前的 SARS 爆发期间使用。 冠状病毒可用于模拟疾病进展并测试疫苗和治疗方法。 非常适合模拟人类呼吸道病毒感染，主要是因为它们在解剖学和结构上的相似性 Chad J. Roy 博士（杜兰国立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联合研究员）最近产生了新颖的转化性成果 通过鼻腔/气管内接种使用 SARS-CoV-2 感染的 NHP 模型，能够 在这项试点探索性研究中，重现了重症 COVID-19 患者的临床快速恶化情况。 我们建议评估两种新型 NHP（恒河猴和非洲绿）模型的尿液转录组学 SARS-2-CoV-2 感染及其与 AKI 早期标志物的关联我们的总体假设是， 因为ACE2可能在COVID-19引起的肾损伤中发挥关键作用，尿转录的变化 特征与尿液早期肾损伤标志物的改变相关，并反映了以下因素的影响： 使用尿液转录组学评估 SARS-CoV-2 对肾脏 ACE2 表达细胞和肾功能的影响。 对从早期感染期间收集的尿液中分离出的表达 ACE2 的细胞进行单细胞 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标：测试 假设尿液转录组学的变化证明了 SARS-CoV2 对肾脏的影响 并伴随着感染 SARS-CoV-2 的 NHP 中肾损伤标记物的早期排泄，具体方式如下： A) 评估早期感染、进展和感染过程中尿肾损伤标志物的时间变化 B) 确定早期肾损伤标志物是否从尿液中排出。 先于肾损伤和 AKI 的临床参数；以及 C) 检查尿液的变化 在 COVID-19 期间以单细胞分辨率对表达 ACE2 的细胞进行转录组学研究取得了成功。 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。