Glomerular Cell-Cell Crosstalk and Injury

肾小球细胞间的串扰和损伤

基本信息

批准号：
10470126
负责人：
Ilse Sofia Daehn
金额：
$ 38.33万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10470126
关键词：
3-Dimensional Adult Affect American Apoptosis Apoptotic Automobile Driving Biological Cells Cellular Stress Chronic Kidney Failure Coculture Techniques Communication Complement Factor B Diabetic Nephropathy Disease Disease Progression Endothelial Cells Endothelium Experimental Models Functional disorder Funding Glomerulonephritis Heat shock proteins Human In Situ In Vitro Inbred BALB C Mice Injury Kidney Lead Lesion Ligands Longitudinal Studies Manuscripts Mediating Microfluidic Microchips Mitochondria Modeling Mus Outcome Oxidative Stress Pathogenesis Pathway interactions Patients Peripheral Phosphotransferases Pre-Eclampsia Prevalence Process Proteins Proteomics Public Health Receptor Signaling Renal Hypertension Renal glomerular disease Role Scheme Sclerosis Signal Transduction Stress Stress Response Signaling Surface System Technology Transforming Growth Factors Transgenic Mice Urine Zebrafish biological adaptation to stress cell injury diagnostic strategy economic impact effective therapy glomerular endothelium glomerulosclerosis in vivo innovation insight mouse model novel novel diagnostics novel therapeutics phenotypic biomarker podocyte preservation receptor release factor response response to injury therapeutic target transcriptomics

项目摘要

Project Summary Chronic kidney disease (CKD) is estimated to affect over 14% of US adults and is increasing in prevalence worldwide. The majority of cases are caused by glomerular diseases with sclerotic lesions, and transforming growth factor β expression in podocytes is a common stress response signal associated with segmental sclerosis. In the previous funding cycle we established a new hypothesis for “glomerular cell-cell crosstalk” using an inducible transgenic mouse model (PodTbrI) that enables podocytes specific and ligand-independent expression of transforming growth factor β type I receptor (TbrI) kinase. In this model, TbrI signaling in podocytes results in the release of Edn1, followed by increased Ednra-mediated mitochondrial oxidative stress and dysfunction of adjacent glomerular endothelial cells (GEC), which, in response, release factor(s) that mediate damage and depletion of adjacent podocytes. This was also demonstrated in other models of experimental podocytopathies (Balb/c + Adryamicin, PodDicerKO mice). We identified that a similar stressed endothelial-to-podocyte crosstalk underlies segmental lesions in DKD. We characterized podocyte mitochondrial dynamics in response to transforming growth factor β signaling. We also performed transcriptomic analysis of isolated GECs after TbrI signaling activation and with this strategy we identified novel GEC injury response pathways. Further, using state of the art proteomics, we identified a panel of novel proteins released by stressed GECs that induce podocyte injury. We also identified key phenotypic markers of cell crosstalk in vivo (GEC ultrastructural changes and loss of glomerular endothelial surface layer). Our findings provide new insights into crosstalk of stressed GECs and podocytes in the pathogenesis of CKD. We hypothesize that the identified GEC secreted proteins mediate podocyte injury and loss in CKD. In this competitive renewal application, we aim to examine and validate the activity of the identified GEC-secreted proteins and cell-cell crosstalk mechanisms mediating podocyte injury in CKD in the following 3 specific aims: SPECIFIC AIMS: 1) To characterize GEC stress and the mechanisms leading to dysfunction and release of crosstalk factors that impact podocytes. 2) To determine GEC response to podocyte activation in vivo. 3) To characterize and validate podocyte responses to GEC secreted factors and determine their functional effects in vitro with a novel 3-D kidney-on-a-chip microfluidic culture system and in vivo. LONG-TERM: The studies proposed in this application will advance our understanding of communications between cells in the glomerulus that underlie the initiation and progression of glomerular disease. The significance of the proposed studies is in the discovery of the requirements and mechanisms for interdependent crosstalk between activated podocytes and stressed endocapillary cells that determine irreversible segmental sclerosis and disease progression. The outcomes will help identify novel diagnostic approaches and potential therapeutic targets for the treatment of glomerular diseases.

项目概要据估计，超过 14% 的美国成年人患有慢性肾病 (CKD)，并且患病率正在上升在世界范围内，大多数病例是由伴有硬化病变和转化性生长的肾小球疾病引起的。足细胞中的β因子表达是与节段性硬化症相关的常见应激反应信号。在上一个资助周期中，我们利用以下方法建立了“肾小球细胞间串扰”的新假设：诱导型转基因小鼠模型 (PodTbrI)，可实现足细胞特异性且不依赖配体转化生长因子 β I 型受体 (TbrI) 激酶的表达在此模型中，TbrI 信号传导。足细胞导致 Edn1 释放，随后增加 Ednra 介导的线粒体氧化应激以及相邻肾小球内皮细胞 (GEC) 的功能障碍，作为反应，GEC 会释放因子介导邻近足细胞的损伤和消耗，这也在其他模型中得到了证明。我们发现实验性足细胞病（Balb/c + 阿德霉素、PodDicerKO 小鼠）存在类似的应激。内皮细胞与足细胞的串扰是 DKD 节段性病变的基础。我们还进行了线粒体动力学响应转化生长因子 β 信号传导。 TbrI 信号激活后分离的 GEC 的转录组分析，通过这种策略，我们确定了此外，利用最先进的蛋白质组学，我们确定了一组新颖的 GEC 损伤反应途径。我们还鉴定了应激 GEC 释放的诱导足细胞损伤的关键表型标记。体内细胞串扰（GEC 超微结构变化和肾小球内皮表面层损失）。我们的研究结果为 CKD 发病机制中应激 GEC 和足细胞的串扰提供了新的见解。我们发现，已鉴定的 GEC 分泌蛋白介导 CKD 中的足细胞损伤和损失。竞争性更新申请，我们的目标是检查和验证已识别的 GEC 分泌的活性介导 CKD 足细胞损伤的蛋白质和细胞间串扰机制有以下 3 个具体目标：具体目标： 1) 表征 GEC 应激以及导致功能障碍和释放的机制影响足细胞的串扰因素 2) 确定 GEC 对足细胞体内激活的反应。表征和验证足细胞对 GEC 分泌因子的反应，并确定它们在使用新型 3D 肾芯片微流体培养系统进行体外实验和体内实验。长期：本申请中提出的研究将增进我们对通信的理解肾小球中导致肾小球疾病发生和进展的细胞之间的关系。拟议研究的意义在于发现了激活的足细胞和受压的毛细血管内细胞之间相互依赖的串扰决定了不可逆节段性硬化症和疾病进展的结果将有助于确定新的诊断方法。治疗肾小球疾病的方法和潜在的治疗靶点。