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Phosphorylation of the podocyte cytoskeleton in diabetic nephropathy

糖尿病肾病足细胞细胞骨架的磷酸化

基本信息

批准号：
10287650
负责人：
Di Feng
金额：
$ 13.13万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-23 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10287650
关键词：
Actinin Actins Address Affect Animal Model Animals Award Biomedical Engineering Cell model Chronic Kidney Failure Collaborations Complications of Diabetes Mellitus Crosslinker Culture Media Cytoskeleton Data Defect Diabetes Mellitus Diabetic Nephropathy Disease Event Exposure to Filtration Focal Segmental Glomerulosclerosis Fostering Functional disorder Funding Future Genetic Glucose Goals Health Care Costs Human Hyperglycemia In Vitro Investigation Kidney Kidney Diseases Knowledge Lead Link Mass Spectrum Analysis Mechanical Stress Mechanics Mediating Mediator of activation protein Mentored Research Scientist Development Award Microfluidics Mission Modeling Morbidity - disease rate Mus Mutation National Institute of Diabetes and Digestive and Kidney Diseases Nature Pathogenesis Pathway interactions Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Play Population Postdoctoral Fellow Public Health Rattus Research Role Sampling Serine Stress Stretching Structure Techniques Technology Testing Tissues United States National Institutes of Health Work alpha Actinin blood filter cost diabetic in vivo in vivo Model innovation insight mortality mutant novel podocyte shear stress

项目摘要

Podocyte dysfunction is an early, key event in the pathogenesis of diabetic nephropathy. Podocytes rely on their cytoskeleton to maintain their structure and function while facing constant mechanical stress inside the glomerulus. a-Actinin 4 (ACTN4) is an essential crosslinker of the actin cytoskeleton; mutations in ACTN4 lead to human kidney disease. Recent evidence has identified an important phosphorylation event in ACTN4 at serine (S) 159. In phosphomimetic cellular and animal models, this phosphorylation of ACTN4 is associated with podocyte vulnerability under mechanical stress. Moreover, phosphorylation of wild type (WT) ACTN4 at S159 is stimulated by high glucose and associated with cytoskeletal derangements, similar to derangements associated with disease-causing mutant ACTN4. The long-term goal that this R03 application advances is to understand the cytoskeleton’s role in podocyte dysfunction underlying diabetic nephropathy. The overall objective of the current proposal is to elucidate the pathway by which high glucose leads to phosphorylation of ACTN4 as a potential mediator of podocyte vulnerability. The central hypothesis is that increased phosphorylation of ACTN4 is stimulated by high glucose and fosters podocyte vulnerability to mechanical stress. The rationale for this project is that finding new pathways leading to podocyte vulnerability could fill critical gaps in knowledge related to the pathogenesis of diabetic nephropathy. To attain the overall objective of this application, the following two specific aims will be pursued. Aim 1 will define the association between phosphorylation of ACTN4 and diabetic nephropathy in vivo. Wild type (WT) control and diabetic nephropathy kidney tissue will be obtained from mice, rats, and humans. Targeted mass spectrometry will be used to quantify ACTN4 phosphorylation across all samples. Aim 2 will determine the impact of high glucose-mediated ACTN4 phosphorylation in vitro. Microfluidic glomeruli-on-chips will be seeded with either human podocytes carrying WT ACTN4 or human podocytes carrying nonphosphorylatable S159A ACTN4. These glomeruli-on-chips will be exposed to culture media containing high glucose while subjected to mechanical stretch and shear stress. The proposed research is innovative since it employs the latest, state-of-the art techniques to study phosphorylation of the cytoskeleton and podocyte vulnerability. The proposed research is significant because it will define a novel pathway by which high glucose mediates podocyte dysfunction through phosphorylation of the podocyte cytoskeleton. Demonstrating that (1) this pathway is upregulated in in vivo models of diabetic nephropathy and that (2) this pathway contributes to podocyte vulnerability under mechanical stress will provide strong justification to further study phosphorylation of ACTN4 as a mechanism underlying the onset and progression of diabetic nephropathy. Data from this R03 will support an R01 to identify the kinases and/or phosphatases that regulate phosphorylation of ACTN4. This future work will not only provide new mechanistic insights into podocyte dysfunction involved in diabetic nephropathy, but also potential targets for novel treatments that mitigate podocyte dysfunction.

足细胞功能障碍是糖尿病肾病发病的早期关键事件。足细胞依赖于它们的