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。

• 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
• 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' 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

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.

在理解人类肾脏疾病进展的机制方面存在根本性的差距 疾病长期目标是表征调节肿瘤进展的关键细胞和分子途径。 急性和慢性肾脏疾病（阿基和CKD），以确定评估疾病进展的新标志物， 并开发针对这些途径的特定治疗干预措施。管间串扰 肾脏中的亚段和免疫细胞是纤维化和疾病进展的重要决定因素。 因此，本申请的目的是选择性地检查肾小管上皮细胞的转录组。 以定量和定位与肾小管亚段和肾动脉亚段相关的免疫细胞亚型。 阿基和CKD患者的结构。本申请的中心假设是， 肾小管亚段和免疫细胞亚型的丰度和分布是独特的， 人类肾脏疾病疾病进展的补充标识符。建议的理由 研究表明，一旦与疾病进展相关的独特分子和细胞标识符， 确定长期结果，它们可用于监测药物干预的功效， 确定最能代表人类疾病的动物模型用于转化研究，并揭示新的 治疗这些疾病的方法。中心假设将通过追求三个具体的测试 目的：1）确定阿基患者活检组织中肾小管亚段表达的转录组 和CKD，并将分子标记与临床结果相关联。2)确定丰度， 免疫细胞亚型在同一组患者活检中的分布。3)Backmap关键分子通路 以确定分子、细胞和结构的相关性。在第一个目标下，激光 将对来自病例匹配的生物库肾活检组织进行小管亚段的显微切割。 快速进展的阿基和CKD患者和未快速进展的患者。基因表达 将对从小管亚段分离的RNA进行分析，以辨别转录组学特征。 管状子段的签名。在第二个目标下，先进的三维组织细胞术 将对来自第一次报告中描述的相同病例匹配患者的生物库肾活检进行 目的是量化免疫细胞组成，检查空间细胞组织，并描绘详细的 快速进展和未快速进展的患者的形态学差异。第三 目的是，我们将对关键分子通路进行染色，以创建人体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疾病病理生理学的重要性， 进展这项拟议中的研究意义重大，因为它是一系列研究的下一步， 预计将确定急需的疾病进展生物标志物，优化临床前研究， 针对阿基和CKD这一巨大的临床问题，开发特异性和针对性的治疗干预措施。