Glucose-Induced Stiffening of Extracellular Matrix in Diabetic Nephropathy

糖尿病肾病中葡萄糖诱导的细胞外基质硬化

基本信息

批准号：
9380480
负责人：
Nicholas Ferrell
金额：
$ 7.9万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-03 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9380480
关键词：
Advanced Glycosylation End Products Affect Basement membrane Binding Biochemical Biomedical Engineering Cell Culture Techniques Cell Proliferation Cell Survival Cells Characteristics Chronic Kidney Failure Collagen Cytoskeleton Development Diabetes Mellitus Diabetic Nephropathy Dialysis procedure Disease Disease Progression Environment Epithelial Epithelial Cells Exposure to Extracellular Matrix Extracellular Matrix Proteins Family Glomerular Mesangial Cell Glucose Goals Homeostasis Hyperglycemia In Vitro Incubated Integrin Binding Integrin alpha1 Integrin alpha2 Integrins Kidney Kidney Diseases Kidney Failure Kidney Glomerulus Knockout Mice Left Ligands Link Liver diseases Malignant neoplasm of lung Mass Spectrum Analysis Measurement Measures Mechanics Mediating Mediator of activation protein Methods Modeling Modification Pathway interactions Phenotype Physiological Predisposition Production Property Protein Isoforms Public Health Renal tubule structure Ribose Role Signal Transduction Structure Techniques Testing Therapeutic Intervention Tissues Tubular formation United States base cell behavior cell type crosslink diabetic extracellular feeding glomerular basement membrane glucose metabolism glycation in vitro Model in vivo inhibitor/antagonist insight interstitial mesangial cell migration mouse model prevent receptor receptor for advanced glycation endproducts response sugar

项目摘要

The goal of this project is to determine how changes in kidney extracellular matrix (ECM) stiffness caused by advanced glycation impact progression of diabetic nephropathy. Our overarching hypothesis is that advanced glycation end products (AGEs) form crosslinks that increase ECM stiffness in the kidney and alter integrin signaling to trigger increased extracellular matrix synthesis. The newly synthesized matrix is further crosslinked, thus creating a feed forward cycle of increased stiffness and ECM synthesis. We propose that this mechanism contributes to accumulation of ECM in the glomerulus and tubulointerstitium that is characteristic of diabetic kidney disease. Our preliminary studies show that incubating renal tubules with glucose or ribose significantly increases the stiffness of the tubular basement membrane. The aims of this application are to (1) Determine the mechanism by which sugar exposure alters the stiffness of kidney tubular and glomerular extracellular matrix and (2) Determine the mechanism by which increased stiffness alters integrin signaling and leads to increased ECM production. The methods employed in aim 1 will include measurement of tubular basement membrane stiffness in normal and sugar modified kidney tubules using our newly developed microcantilever-based stiffness measurement technique. Glomerular stiffness will be measured using microscale compression testing. Mechanical characterization of the tissue will be correlated with biochemical analysis via mass spectrometry to evaluate crosslink formation. For aim 2, we will use in vitro cell culture models that mimic the mechanical properties of normal and sugar modified tubular and glomerular ECM. Stiffness induced changes in cell phenotype, cell proliferation and ECM production will be evaluated in wild type and integrin α1 and α2-null tubular epithelial and glomerular mesangial cells. Successful completion of these aims will establish the role of AGE- mediated ECM stiffening as a potential contributor to progression of diabetic kidney disease and will inform subsequent studies in diabetic and integrin-null mouse models.

该项目的目标是确定肾脏细胞外基质 (ECM) 硬度的变化是如何引起的晚期糖化影响糖尿病肾病的进展。糖化终末产物 (AGE) 形成交联，增加肾脏 ECM 硬度并改变整合素信号触发细胞外基质合成增加，新合成的基质进一步交联，从而创建增加刚度和 ECM 合成的前馈循环。导致肾小球和肾小管间质中 ECM 的积累，这是糖尿病的特征我们的初步研究表明，用葡萄糖或核糖孵育肾小管会显着降低肾脏疾病的风险。增加管状基底膜的刚度该应用的目的是 (1) 确定糖暴露改变肾小管和肾小球细胞外基质硬度的机制 (2) 确定增加的硬度改变整合素信号并导致增加的机制目标 1 中采用的 ECM 生产方法将包括管状基底膜的测量。使用我们新开发的基于微悬臂梁的刚度来测量正常肾小管和糖修饰肾小管的刚度肾小球硬度将使用微尺度压缩测试来测量。组织的机械特性将通过质谱与生化分析相关联，以对于目标 2，我们将使用模拟机械的体外细胞培养模型来评估交联形成。正常和糖修饰的肾小管和肾小球 ECM 的特性诱导细胞变化。将在野生型和整合素 α1 和 α2 缺失管状细胞中评估表型、细胞增殖和 ECM 产生上皮细胞和肾小球系膜细胞的成功完成将确立 AGE- 的作用。介导的 ECM 硬化是糖尿病肾病进展的潜在因素，并将告知糖尿病和整合素缺失小鼠模型的后续研究。