Nephron Sub-segmental Omics and Quantitative 3D Imaging of Human Kidney.

人类肾脏的肾单位亚节段组学和定量 3D 成像。

基本信息

批准号：
10242708
负责人：
Tarek Maurice Ashkar
金额：
$ 67.91万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2022-09-19
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10242708
关键词：
3-Dimensional Acute Acute Renal Failure with Renal Papillary Necrosis Adopted Animal Model Atlases Back Biological Markers Biopsy Biopsy Specimen Caring Cells Cellular Structures Chronic Kidney Failure Clinical Core Biopsy Cytometry Data Development Disease Disease Progression Duct (organ) structure Epigenetic Process Evaluation Fibrosis Functional disorder Future Gene Expression Profile Gene Expression Profiling Goals Health Health system Henle&apos s loop Human Image Immune Indiana Injury Intervention Kidney Kidney Diseases Knowledge Lasers Maps Methodology Methods Microdissection Mission Molecular Molecular Analysis Molecular Profiling Monitor Morphology National Institute of Diabetes and Digestive and Kidney Diseases Natural regeneration Nephrons Outcome Pathogenesis Pathology Pathway interactions Patients Pharmacology Phase Phenotype Physical shape Proteomics Public Health Quality of life RNA Research Software Tools Spatial Distribution Stains Structure Techniques Testing Therapeutic Therapeutic Intervention Three-Dimensional Imaging Tissues Translating Translational Research Tubular formation United States National Institutes of Health Vascular Endothelium biobank cell type clinical practice cohort density digital digital models fluorescence imaging follow-up human disease human imaging improved injured interstitial cell kidney biopsy molecular phenotype morphometry next generation novel novel marker novel strategies outcome prediction precision medicine preclinical study predict clinical outcome predictive signature repository targeted treatment therapeutic target tool transcriptome transcriptomics

项目摘要

There is a fundamental gap in understanding the mechanisms that determine progression in human kidney disease. The long term goal is to characterize key cellular and molecular pathways regulating progression of acute and chronic kidney diseases (AKI and CKD), to identify novel markers that assess disease progression, and to develop specific therapeutic interventions targeting these pathways. The cross-talk between tubular subsegments and immune cells in the kidney is an important determinant of fibrosis and disease progression. Consequently, the objective of this application is to selectively examine the transcriptome of tubular subsegments and to quantify and localize immune cell subtypes in relation to tubular subsegments and renal structures in patients with AKI and CKD. The central hypothesis of this application is that the transcriptome of kidney tubular subsegments and the abundance and distribution of immune cell subtypes are unique and complimentary identifiers of disease progression in human kidney diseases. The rationale for the proposed research is that once the unique molecular and cellular identifiers that correlate with disease progression and long term outcomes are determined, they can be used to monitor efficacy of pharmacologic interventions, identify animal models that best represent human disease for translational research, and reveal novel approaches towards treating these conditions. The central hypothesis will be tested by pursuing three specific aims: 1) Define the transcriptome expressed by the tubular subsegments from biopsies of patients with AKI and CKD and correlate molecular signatures with clinical outcome. 2) Determine the abundance and distribution of immune cell subtypes in the same set of patient biopsies. 3) Backmap key molecular pathways to the renal biopsy in order to define molecular, cellular and structural correlations. Under the first aim, laser microdissection of tubular subsegments will be performed on biobanked kidney biopsies from case-matched patients with AKI and CKD that rapidly progressed and those that did not rapidly progress. Gene expression analysis will be performed on RNA isolated from the tubular subsegments to discern the transcriptomic signature of tubular subsegments. Under the second aim, advanced three-dimensional (3-D) tissue cytometry will be performed on the biobanked kidney biopsies from the same case-matched patients described in the first aim to quantify the immune cell composition, examine spatial cellular organization, and delineate detailed morphologic differences in patients who rapidly progressed and those who did not rapidly progress. In the third aim, we will stain for key molecular pathways in order to create a large-scale digital model of the human biopsy that highlights molecular, cellular and structural correlations important for disease pathophysiology and progression. The proposed research is significant, because it is the next step in a continuum of research that is expected to identify critically needed biomarkers of disease progression, optimize preclinical studies, and develop specific and targeted therapeutic interventions in the vast clinical problem of AKI and CKD.

理解决定人肾脏进展的机制存在根本的差距疾病。长期目标是表征调节调节进展的钥匙细胞和分子途径急性和慢性肾脏疾病（AKI和CKD），以确定评估疾病进展的新标记，并开发针对这些途径的特定治疗干预措施。管状之间的串扰肾脏中的亚播种和免疫细胞是纤维化和疾病进展的重要决定因素。因此，该应用的目的是选择性检查管状的转录组与管状亚况和肾脏相关的亚播种并定位和定位免疫细胞亚型 AKI和CKD患者的结构。该应用的中心假设是肾小管的亚型以及免疫细胞亚型的丰度和分布是独特的，并且人类肾脏疾病中疾病进展的免费鉴定。提议的理由研究是，曾经曾经是与疾病进展相关的独特分子和细胞标识符确定长期结果，可以用于监测药理学干预措施的功效，确定最能代表人类疾病的动物模型，并揭示新颖的动物模型治疗这些疾病的方法。中心假设将通过追求三个特定的特定目的：1）定义由AKI患者活检的管状亚播种表达的转录组和CKD并将分子特征与临床结果相关联。 2）确定丰度和免疫细胞亚型在同一组患者活检中的分布。 3）反向图钥匙分子途径进行肾脏活检，以定义分子，细胞和结构相关性。在第一个目标下，激光小管状子播种的显微减退将在病例匹配的生物群肾脏活检中进行 AKI和CKD的患者迅速进展，并且没有快速进展的患者。基因表达分析将对从管状亚播种分离的RNA进行，以辨别转录组管状亚况的特征。在第二个目标下，高级三维（3-D）组织细胞术将在第一个中描述的同一病例匹配的患者的生物群肾活检中进行旨在量化免疫细胞组成，检查空间细胞组织并详细描述迅速发展的患者和未迅速进展的患者的形态差异。在第三目的，我们将为关键分子途径染色，以创建人类活检的大规模数字模型这突出了分子，细胞和结构相关性对于疾病病理生理学和进展。拟议的研究很重要，因为这是一项连续研究的下一步预计将确定疾病进展的急需的生物标志物，优化临床前研究，并在AKI和CKD的巨大临床问题中开发特定和有针对性的治疗干预措施。