Novel regulatory mechanisms of the glomerular endothelium

肾小球内皮的新调节机制

基本信息

批准号：
10621339
负责人：
JANOS PETI-PETERDI
金额：
$ 44.87万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-03 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621339
关键词：
Adult Affect Albumins Albuminuria Angiogenic Factor Angiogenic Proteins Animal Model Biological Assay Blood Blood Vessels Brain Cardiovascular Diseases Cell Culture Techniques Cell Line Cell Lineage Cells Chief Cell Chronic Kidney Failure Code Darkness Development Disease Disease Progression Disease model Endothelial Cells Endothelium Enzyme-Linked Immunosorbent Assay Equilibrium Extravasation Fibrosis Fibrous capsule of kidney Foundations Functional disorder Genes Genetic Glomerular Capillary Health Heart Histology Hormones Implant In Vitro Injury Juxtaglomerular Apparatus Juxtaglomerular Cell Kidney Kidney Diseases Knock-out Knockout Mice Knowledge Laboratories Macula densa Maintenance Measurement Mediating Methods Mus Natural regeneration Nephrectomy PTGS2 gene Paracrine Communication Pathogenicity Pathology Phenotype Physiological Plasma Play Population Production Regulation Research Role Signal Transduction Sodium Chloride Structure Technology Testing Tissues Transgenic Mice Visual Visualization WNT Signaling Pathway angiogenesis cardiovascular health cell injury cell type comorbidity diabetic diagnostic strategy dietary salt gain of function glomerular endothelium hypertensive imaging approach implantation improved in vivo intravital imaging loss of function mortality mouse model multiphoton microscopy novel novel diagnostics novel therapeutics paracrine pharmacologic precursor cell programs regenerative therapy repaired response salt intake stem cells uptake

项目摘要

Glomerular endothelial cells play pivotal roles in the maintenance of the normal structure and function of the healthy glomerulus, and also in the development of glomerular injury in kidney diseases. However, our knowledge of the mechanisms that constantly maintain and regenerate the glomerular endothelium after injury have been limited, due to the inaccessibility of this cell type and lack of in vivo research technologies. Our laboratory has recently developed novel transgenic mouse models and an intravital imaging approach using serial multiphoton microscopy (MPM) of the same glomerulus over several days-weeks to quantitatively visualize and track the fate and function of the same single glomerular endothelial cells in the intact living kidney. This technical advance provided visual clues on the development of clonal endothelial cell clusters at the glomerular vascular pole in response to dietary salt restriction and various types on glomerular injury, and suggested its control by the juxtaglomerular apparatus (JGA). In addition, the foundation of this proposal is our recent preliminary findings which suggested that macula densa (MD) cells of the JGA are novel chief organizers and master regulators of glomerular remodeling in the adult kidney. Gene profiling showed that MD cells produce novel secreted tissue remodeling factors that act in a paracrine fashion on resident progenitor cells. One of the top new MD genes responsible for this new glomerular remodeling program was identified as CCN1 (also known as Cyr61), coding for a secreted angiogenic protein that is known to target endothelial cells as well as other cell types. Our main hypothesis is that MD cells, via Wnt signaling-mediated synthesis and secretion of angiogenic proteins including CCN1, are major paracrine regulators of the glomerular endothelium. This project will use comprehensive experimental approaches including in vitro cell culture, new transgenic mouse models, fluorescently tagged cell lineages, cell fate tracking, in vivo disease models and serial MPM to investigate the angiogenic role, mechanisms, and importance of MD cells and CCN1 in glomerular endothelial maintenance and repair. The specific aims are to (i) Examine the role and mechanism of MD cells in the maintenance of normal glomerular capillary structure and function, (ii) Study the regulation of the MD cell angiogenesis promoting program, and (iii) Test for renal and cardiovascular disease modifying roles and mechanisms of MD cells.

肾小球内皮细胞在维持正常结构和功能中发挥着关键作用健康肾小球的影响，以及肾脏疾病中肾小球损伤的发展。然而，我们对肾小球持续维持和再生机制的了解由于这种细胞类型难以接近且体内缺乏研究技术。我们的实验室最近开发了新型转基因小鼠模型和使用连续多光子显微镜（MPM）对同一肾小球进行活体成像几天到几周的时间来清晰地可视化和跟踪同一定量单的命运和功能完整的活肾中的肾小球内皮细胞。这一技术进步提供了视觉线索肾小球血管极克隆内皮细胞簇的发育对饮食的反应。盐限制和各种类型的肾小球损伤的关系，并提出其通过肾小球旁的控制此外，该提案的基础是我们最近的初步发现，其中表明 JGA 的致密斑 (MD) 细胞是新的主要组织者和主要调节者成人肾脏肾小球重塑的基因分析表明 MD 细胞产生新的。分泌的组织重塑因子以旁分泌方式作用于常驻祖细胞之一。负责这一新肾小球重塑程序的顶级新 MD 基因被确定为 CCN1 （也称为 Cyr61），编码一种已知靶向内皮细胞的分泌性血管生成蛋白以及其他细胞类型。我们的主要假设是 MD 细胞通过 Wnt 信号介导的合成和包括 CCN1 在内的血管生成蛋白的分泌，是肾小球的主要旁分泌调节因子该项目将使用包括体外细胞在内的综合实验方法。培养，新转基因小鼠模型，荧光标记细胞谱系，细胞命运追踪，体内疾病模型和系列 MPM 来研究 MD 的血管生成作用、机制和重要性细胞和 CCN1 在肾小球内皮维护和修复中的作用。具体目标是 (i) 检查。 MD细胞在维持正常肾小球毛细血管结构中的作用和机制功能，(ii) 研究 MD 细胞血管生成促进程序的调节，以及 (iii) 肾脏测试。和心血管疾病改变 MD 细胞的作用和机制。