Single-cell analysis to promote kidney repair

单细胞分析促进肾脏修复

基本信息

批准号：
10646473
负责人：
BENJAMIN D. HUMPHREYS
金额：
$ 73.22万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646473
关键词：
3-Dimensional Ablation Acute Acute Renal Failure with Renal Papillary Necrosis Adopted Atlases Attenuated Automobile Driving Bioinformatics California Candidate Disease Gene Cell Nucleus Cells Chromatin Chronic Disease Chronic Kidney Failure Communities Data Data Set Databases Development Diagnosis Epigenetic Process Epithelial Cells Excision Female Fibrosis Gene Deletion Gene Expression Genes Genetic Genetic Transcription Hospitalization Human Immunohistochemistry Impairment In Situ Hybridization Inflammation Inflammatory Injury Injury to Kidney Intervention Kidney Kidney Diseases Knock-out Knowledge Laboratories Logic MAP4K4 gene Maps Medicare Modeling Modification Molecular Mus NF-kappa B Nephrons Nuclear Nuclear RNA Online Systems Outcome Pathway interactions Patient-Focused Outcomes Patients Persons Population Predisposition Probability Process Profibrotic signal Property Recovery Recurrence Regulator Genes Renal function Renal tubule structure Research Resources Risk Role Route Sampling Sex Differences Signal Transduction Surveys TNF Receptor-Associated Factors Technology Testing Therapeutic Intervention Time Transgenic Mice Tubular formation Universities Visualization software Washington adverse outcome aged cell type data mining epigenome epithelial injury experience genetic approach high risk improved improved outcome injured kidney biopsy kidney repair male member men mouse model multimodal data multimodality multiple omics novel novel strategies novel therapeutic intervention prevent profibrotic cytokine repair strategy repaired response response to injury sex single cell analysis single molecule single nucleus RNA-sequencing single-cell RNA sequencing therapeutic development therapeutic target transcriptome transcriptome sequencing transcriptomics

项目摘要

Summary Acute kidney injury (AKI) has a wide spectrum of outcomes from recovery to a long-term transition to chronic kidney disease (CKD). Between 2000 and 2014, AKI hospitalizations have increased from 3.5 to 11.7 per 1000 persons. Medicare patients aged 66 years and older hospitalized for AKI have a 35% cumulative probability of a recurrent AKI hospitalization within one year and 28% will be diagnosed with CKD in the same time frame. Men have a higher risk of AKI, and of developing progressive CKD, although the mechanisms are poorly understood. In the mouse, males also show a heightened vulnerability to AKI. Recent single cell RNA-seq studies from the McMahon and Kim groups have highlighted marked differences in gene expression between the sexes in proximal tubule segments, the region of the nephron most susceptible to AKI. Preliminary studies in the Humphreys and McMahon laboratories using single nuclear sequencing identified a cell type resulting from failed repair of proximal tubule cells (FR-PTC) following mild to severe AKI with a pro-inflammatory, pro- fibrotic signature. FR-PTCs are hypothesized to drive progressive kidney disease following AKI. This proposal centers on the postulates that an understanding of sex differences in response to AKI, and the application of genetic approaches to target proinflammatory properties of FR-PTCs and to eliminate FR-PTCs following renal repair, will be effective routes to ultimately benefit patient outcomes post AKI. To this end, we have assembled a complementary team, with prior collaborative experience: Humphreys (Washington University), Kim (University of Pennsylvannia) and McMahon (University of Southern California). All team members have participated in the ReBuilding a Kidney Consortium. In Specific Aim 1: we will characterize successful versus failed proximal tubule repair with single nucleus transcriptomics (snRNA-seq) and single nuclear chromatin accessibility studies (scATAC-seq) in male and female mouse models examining key findings in human kidney biopsies. In Specific Aim 2: we will harmonize multimodal datasets generated in Specific Aim1 to facilitate viewing and interrogation of these data by the broad research community. Mining of these data by the group will focus on defining the regulatory logic of repair strategies and outcomes in the male and female kidney. In Specific Aim 3: we will examine the hypothesis that adverse outcomes in the male kidney following AKI are driven by NF-kB pathway components Nfkb1 and TNIK in FR-PTCs, genetically eliminating the action of these genes. We will generate and validate a new transgenic mouse resource for the community, enabling genetic modification and elimination of FR-PTCs. We will determine whether FR-PTC removal has a favorable outcome, as we predict, on progressive kidney disease following AKI.

概括从恢复到长期过渡到慢性的急性肾脏损伤（AKI）具有广泛的结果肾脏疾病（CKD）。在2000年至2014年之间，AKI住院治疗已从3.5升至11.7 人。 66岁及以上的Medicare患者住院的AKI患者的累积概率为35％一年内复发性的AKI住院，将在同一时间范围内被诊断出患有CKD。男性患AKI的风险较高，并且有进步的CKD，尽管这些机制很差理解。在鼠标中，男性还显示出对AKI的脆弱性。最近的单细胞RNA-seq 麦克马洪和金体组的研究突出了基因表达之间的明显差异近端小管段中的性别，肾肾脏最容易受到AKI的影响。初步研究在使用单个核测序的汉弗莱斯和麦克马洪实验室中，确定了导致的细胞类型在轻度至重度AKI之后的近端小管细胞（FR-PTC）的修复失败中纤维化特征。假设FR-PTC在AKI之后驱动了进行性肾脏疾病。这个建议以假定的理解对AKI的性别差异的理解以及应用靶向FR-PTC的促炎特性并消除肾脏后FR-PTC的遗传方法维修将是有效的途径，最终使AKI后患者结果受益。为此，我们已经组装了一个互补的团队，具有先前的合作经验：汉弗莱斯（华盛顿大学），金（宾夕法尼亚大学）和麦克马洪（南加州大学）。所有团队成员都有参加了重建肾脏财团。在特定目标1中：我们将表征成功与单核转录组学（SNRNA-SEQ）和单核染色质的近端小管修复失败雄性和女鼠模型中可及性研究（SCATAC-SEQ）检查人类肾脏中的关键发现活检。在特定目标2中：我们将协调特定AIM1中生成的多模式数据集以促进广泛的研究社区查看和询问这些数据。小组挖掘这些数据将重点放在定义男性和雌性肾脏修复策略和结果的调节逻辑上。在特定目的3：我们将研究以下假设：AKI之后的雄性肾脏不良结果是在FR-PTC中，由NF-KB途径组件NFKB1和TNIK驱动，从遗传上消除了这些作用基因。我们将为社区生成并验证新的转基因鼠标资源，使遗传修改和消除FR-PTC。我们将确定FR-PTC删除是否有利正如我们预测的那样，结果是AKI后进行性肾脏疾病。