Mechanisms of Kidney Injury in COVID-19

COVID-19 肾损伤的机制

基本信息

批准号：
10483130
负责人：
Shreeram Akilesh
金额：
$ 40.81万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-07 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10483130
关键词：
2019-nCoV APOL1 gene Acute Renal Failure with Renal Papillary Necrosis Address Affect Alleles Anatomy Autopsy Bacterial Artificial Chromosomes Biological Models Black race COVID-19 COVID-19 pandemic COVID-19 patient Cells Cellular Morphology Cessation of life Clinical Data Collaborations Data Diagnosis Diagnostic Disease Electron Microscopy Foundations Functional disorder Future Genomics Genotype Goals Guide prevention HIV Infections Hispanic Human Immunohistochemistry Immunology In Situ Hybridization In Vitro Individual Infection Inflammatory Injury Injury to Kidney Interferon Type II Kidney Kidney Diseases Laboratory Animal Models Lung infections Mediating Modeling Molecular Morphology Mus Organoids Pathogenesis Pathologist Pathology Patients Phenotype Physiological Population Heterogeneity Pre-Clinical Model Prevention Prognosis Proteomics Reagent Renal function Research Research Personnel SARS-CoV-2 infection Slice Source Specimen System Therapeutic Intervention Tissues Transgenic Mice Viral Virus Virus Diseases biobank cell type cytokine cytotoxicity defined contribution experimental study high risk human tissue in vitro Model in vivo induced pluripotent stem cell kidney biopsy kidney cell kidney infection model development mortality risk mouse genetics mouse model prevent response risk variant transcriptomics

2019-nCoV APOL1 gene Acute Renal Failure with Renal Papillary Necrosis Address Affect Alleles Anatomy Autopsy Bacterial Artificial Chromosomes Biological Models Black race COVID-19 COVID-19 pandemic COVID-19 patient Cells Cellular Morphology Cessation of life Clinical Data Collaborations Data Diagnosis Diagnostic Disease Electron Microscopy Foundations Functional disorder Future Genomics Genotype Goals Guide prevention HIV Infections Hispanic Human Immunohistochemistry Immunology In Situ Hybridization In Vitro Individual Infection Inflammatory Injury Injury to Kidney Interferon Type II Kidney Kidney Diseases Laboratory Animal Models Lung infections Mediating Modeling Molecular Morphology Mus Organoids Pathogenesis Pathologist Pathology Patients Phenotype Physiological Population Heterogeneity Pre-Clinical Model Prevention Prognosis Proteomics Reagent Renal function Research Research Personnel SARS-CoV-2 infection Slice Source Specimen System Therapeutic Intervention Tissues Transgenic Mice Viral Virus Virus Diseases biobank cell type cytokine cytotoxicity defined contribution experimental study high risk human tissue in vitro Model in vivo induced pluripotent stem cell kidney biopsy kidney cell kidney infection model development mortality risk mouse genetics mouse model prevent response risk variant transcriptomics

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项目摘要

PROJECT SUMMARY/ABSTRACT The SARS-CoV-2 pandemic has infected millions of individuals in the US and caused hundreds of thousands of deaths. We and others have shown that COVID-19 is also strongly associated with devastating and usually rare kidney pathophysiologies, such as collapsing glomerulopathy (CG). As in HIV infection, CG in COVID-19 patients mostly affects individuals with high-risk APOL1 genotypes, which are more prevalent in Black and some Hispanic patients. To guide treatment, there is a pressing need to understand whether COVID-19 nephropathy is due to direct viral infection or indirect mechanisms, such as cytokines or physiologic disturbances that emanate from the lung infection. Addressing this need has been hampered by poorly validated reagents, and misinterpretation of immunohistochemistry and electron microscopy findings. We have assembled a multi-investigator team to uncover the mechanisms of kidney injury due to SARS-CoV-2 infection. We will bring expert and complementary expertise in anatomic, autopsy and renal pathology, integrative genomic analysis, human kidney organoid systems and mouse immunology. We will use primary human tissue specimens, in vitro human kidney model systems and a new mouse model of COVID-19 to define direct and indirect mechanisms of SARS- CoV-2 associated kidney injury in three specific aims. Aim 1: Using kidney tissue specimens from COVID- 19 patients and controls, we will define the spectrum of kidney manifestations in individuals that have been infected with SARS-CoV-2. We will use immunohistochemistry, in situ hybridization, and proteomics to define SARS-CoV-2 kidney infection in a diverse population. In patients with COVID-19 associated CG, we will define molecular changes of this disease using spatial transcriptomic profiling, and the association with APOL1 status. These studies will define the relationship of SARS-CoV-2 infection to COVID-19 associated kidney diseases and uncover molecular mechanisms that underlie direct and indirect modes of kidney injury. Aim 2: Human kidney organoids provide a physiologically relevant model of SARS-CoV-2 infection. We will define cellular, morphologic and molecular hallmarks of SARS-CoV-2 infection in human kidney organoids and organotypic tissue slices. Using established iPSC cells with APOL1 high-risk alleles, we will determine the impact of APOL1 genotype on infection and inflammatory cytokine induced kidney injury. These studies will establish which kidney cells are capable of being infected by SARS-CoV-2, and kidney cell type specific molecular changes induced by viral infection and inflammatory cytokines. Aim 3: We will use a recently developed mouse adapted SARS-CoV-2 virus to determine the effect of SARS CoV-2 infection on kidney function in vivo. Using newly described BAC- transgenic mice that express human APOL1 G0, G1 or G2 alleles, we will define the influence of human APOL1 high-risk alleles on kidney function and kidney injury during SARS-CoV-2 infection. The successful development of these models will establish a paradigm for investigating viral infection associated kidney injury and leverage mouse genetics to define mechanisms of kidney injury and CG, and well as future therapeutic interventions.

项目摘要/摘要 SARS-COV-2大流行已感染了美国数百万个人，并造成了数十万个死亡人数。我们和其他人已经表明，Covid-19也与毁灭性且通常很少见肾脏病理生理学，例如崩溃的肾小球病（CG）。与HIV感染一样，COVID-19患者的CG 主要影响具有高风险APOL1基因型的人，这些基因型在黑色和一些西班牙裔中更为普遍患者。为了指导治疗，需要了解covid-19肾病是否是由于直接病毒感染或间接机制，例如细胞因子或生理障碍肺部感染。解决这一需求已受到验证不佳的试剂和误解的阻碍免疫组织化学和电子显微镜发现。我们已经组装了一个多启示的团队发现由于SARS-COV-2感染引起的肾脏损伤机制。我们将带来专家和补充解剖学，尸检和肾脏病理学，综合基因组分析，人类肾脏器官的专业知识系统和小鼠免疫学。我们将使用原代人体组织标本，体外人类肾脏模型系统和COVID-19的新小鼠模型，以定义SARS-的直接和间接机制 COV-2在三个特定目标中相关的肾脏损伤。 AIM 1：使用来自Covid-的肾脏组织标本 19例患者和对照组，我们将定义已经感染SARS-COV-2。我们将使用免疫组织化学，原位杂交和蛋白质组学来定义 SARS-COV-2肾脏感染在多样化的人群中。在COVID-19相关CG的患者中，我们将定义该疾病的分子变化使用空间转录组分析以及与APOL1状态的关联。这些研究将定义SARS-COV-2感染与COVID-19的相关肾脏疾病的关系和发现肾损伤直接和间接模式的分子机制。目标2：人类肾脏器官提供了与SARS-COV-2感染的生理相关模型。我们将定义细胞形态学人类肾脏器官和器官组织切片中SARS-COV-2感染的分子标志。使用已建立的IPSC细胞具有APOL1高风险等位基因，我们将确定APOL1基因型对感染和炎症细胞因子诱导肾脏损伤。这些研究将确定哪些肾细胞是能够被SARS-COV-2感染，以及病毒诱导的肾细胞类型特异性分子变化感染和炎症细胞因子。 AIM 3：我们将使用最近开发的鼠标改编的SARS-COV-2 病毒确定SARS COV-2感染对体内肾功能的影响。使用新描述的BAC- 表达人Apol1 G0，G1或G2等位基因的转基因小鼠，我们将定义人类Apol1的影响 SARS-COV-2感染期间的肾功能和肾脏损伤的高风险等位基因。成功的发展这些模型将建立一个用于研究病毒感染相关肾脏损伤和杠杆作用的范式小鼠遗传学来定义肾脏损伤和CG的机制，以及未来的治疗干预措施。