Podocyte Cell Cycle Regulation after DNA Damage

DNA 损伤后足细胞细胞周期调控

• 批准号: 9324233
• 负责人: Astrid Weins
• 金额: $ 8.88万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9324233
• 关键词: Adhesions; Adriamycin PFS; Affect; Budgets; Cardiac Myocytes; Cell Cycle; Cell Cycle Checkpoint; Cell Cycle Regulation; Cell Death; Cell Line; Cell-Matrix Junction; Cells; Cessation of life; Chronic Kidney Failure; Cicatrix; Cyclin-Dependent Kinases; DNA; DNA Damage; Data; Development; Diagnostic; Dialysis patients; Disease; Disease Progression; Event; Extracellular Matrix; Focal Adhesions; Foot Process; G1/S Transition; Genotoxic Stress; Goals; Histologic; Human; Immunofluorescence Microscopy; In Situ; In Vitro; Injury; K-Series Research Career Programs; Kidney Diseases; LIM Domain; Link; Mediating; Medicare; Microscopy; Mitotic; Modeling; Modernization; Molecular; Mus; Nephrons; Neurons; Pathologic; Pathway interactions; Patients; Pattern; Phenotype; Prognostic Marker; Proliferating; Protein Family; Proteins; Proteinuria; Public Health; Renal Replacement Therapy; Renal glomerular disease; Resolution; Role; Scaffolding Protein; Sclerosis; Severities; Techniques; Testing; Therapeutic; United States; base; genotoxicity; glomerular basement membrane; glomerular filtration; human disease; in vivo; injured; innovation; insight; member; mortality; novel; oxidative DNA damage; podocyte; prevent; response; senescence; tool

Project Summary/Abstract The comprehensive term “podocytopathy” represents a poorly defined diagnostic entity, which includes a wide spectrum of histologic patterns of glomerular disease, all characterized by structural changes to the glomerular filter and proteinuria. Current pathomechanistic insights into this diverse group of diseases are limited, and, with the exception of few promising studies, have not provided substantial therapeutic guidance. Pathologically, podocyte loss remains the single unifying observation and closely correlates with disease progression. Podocyte death has recently gained acceptance as a key pathway to podocyte loss, although the molecular mechanisms remain elusive. Hic-5, a member of the LIM domain family of proteins, is increased in podocytes along the glomerular filtration barrier in human podocytopathies. Distinct from a pure function as a focal adhesion protein, our novel preliminary data suggest a pro-survival role for hic-5 in podocytes following injurious events. Specifically, our findings indicate that hic-5 protects podocytes from cell death by mediating cell cycle control through stabilization of p21 after genotoxic stress in vitro and in vivo, thereby maintaining the structural integrity of the glomerular filtration barrier, ameliorating proteinuria and preventing glomerular scarring. The overarching goal of this proposal is to elucidate the pro-survival role of hic-5 following podocyte injury by studying its effect on cell cycle checkpoint control. Specifically, we aim to: 1) Define on a molecular level how hic-5 mediates changes in the podocyte cell cycle after DNA damage by using a podocyte cell line in vitro and murine Adriamycin nephropathy as a model of genotoxic glomerular injury in vivo; and 2) Examine whether the presence of hic-5 in podocyte foot processes results in the activation of a pro-survival signature in podocytopathies in vivo by use of an innovative super-resolution immunofluorescence microscopy technique to validate our findings in human disease. Understanding the role of hic-5 in preventing podocyte loss by regulating the podocyte cell cycle may guide our efforts to discover pathways to nephron loss, identify prognostic biomarkers and develop targeted podocyte-specific therapies for proteinuric kidney diseases.

项目总结/摘要 综合术语“足细胞病”代表了一个定义不明确的诊断实体，其包括广泛的 肾小球疾病的组织学模式谱，均以肾小球结构变化为特征， 过滤器和蛋白尿。目前对这组不同疾病的病理机制的认识是有限的， 除了少数有希望的研究外，还没有提供实质性的治疗指导。 在病理学上，足细胞丢失仍然是唯一统一的观察结果，并与疾病密切相关 进展最近，足细胞死亡被认为是足细胞损失的关键途径，尽管足细胞死亡是一个非常重要的因素。 分子机制仍然难以捉摸。Hic-5是LIM结构域蛋白家族的一员， 足细胞沿着肾小球滤过屏障。与纯函数不同， 粘着斑蛋白，我们的新的初步数据表明，在足细胞中hic-5的促生存作用， 伤害事件。具体地说，我们的研究结果表明，HIC-5通过介导细胞凋亡来保护足细胞。 通过在体外和体内基因毒性应激后稳定p21来控制细胞周期，从而维持 肾小球滤过屏障的结构完整性，改善蛋白尿和预防肾小球滤过 疤痕本研究的主要目的是阐明hic-5在足细胞凋亡后的促生存作用。 通过研究其对细胞周期检查点控制的影响来研究损伤。具体来说，我们的目标是：1）定义一个分子 水平hic-5如何介导DNA损伤后足细胞细胞周期的变化，通过使用足细胞细胞系， 体外和小鼠阿霉素肾病作为体内遗传毒性肾小球损伤模型;和2）检查 足细胞足突中HIC-5的存在是否导致促存活信号的激活， 通过使用创新的超分辨率免疫荧光显微镜技术， 证实了我们在人类疾病中的发现理解hic-5在预防足细胞损失中的作用， 调节足细胞的细胞周期可能会指导我们努力发现肾单位损失的途径， 预后生物标志物，并开发针对蛋白尿肾病的靶向足细胞特异性疗法。