PP2A as a drug target for diabetic kidney disease

PP2A作为糖尿病肾病的药物靶点

• 批准号: 10627834
• 负责人: John Cijiang He
• 金额: $ 58.95万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-21 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10627834
• 关键词: Actins; Adhesions; Affect; Affinity; Albuminuria; Attenuated; Binding; Binding Proteins; Biological Assay; Biology; Cell Adhesion; Cells; China; Chinese Herbs; Chronic Kidney Failure; Cytoskeleton; Data; Development; Diabetes Mellitus; Diabetic Nephropathy; Diabetic mouse; Disease; Disease Progression; Docking; Drug Combinations; Drug Targeting; F-Actin; Genes; Glucose; Herbal Medicine; Human; In Vitro; Inflammation; Inflammatory; Injury; Injury to Kidney; Insulin-Dependent Diabetes Mellitus; Kidney; Kidney Diseases; Knock-out; Knockout Mice; Mass Spectrum Analysis; Mediating; Methods; Modeling; Mus; Mutation; NF-kappa B; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Paper; Pathogenesis; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Phosphorylation; Phosphorylation Site; Play; Protein Dephosphorylation; Protein Isoforms; Protein phosphatase; Proteins; Proteinuria; Regulation; Role; Scanning; Signaling Protein; Site; Streptozocin; Surface Plasmon Resonance; Techniques; Testing; Transgenic Organisms; Variant; Western Blotting; arctigenin; db/db mouse; diabetic; differential expression; drebrins; effective therapy; improved; insight; kidney cell; knock-down; migration; mouse model; mutant; novel; novel drug class; novel therapeutics; p65; phenome; podocyte; protective effect; protein activation; protein expression; protein function; protein phosphatase 2A regulatory subunit 65 kDa; scaffold; transcriptome sequencing; transcriptomics; type I and type II diabetes

Title: Role of PP2A in podocyte biology and diabetic kidney disease SUMMARY Diabetic kidney disease (DKD) remains a leading cause of chronic kidney disease with limited treatment options. Arctigenin (ATG) is a major component derived from the extracts of Fructus Arctii, a traditional Chinese herbal remedy that has shown to confer renoprotection and to reduce proteinuria in patients with DKD. Our preliminary data show that ATG administration alone is sufficient to attenuate proteinuria and podocyte injury in mouse models of type 1 and type 2 diabetes. Transcriptomic analysis of isolated glomeruli from the diabetic and control mice showed that the major pathways affected by ATG treatment were of cell adhesion and inflammation. ATG improved cell adhesion and inhibited migration in cultured human podocytes. By combining the Drug Affinity Responsive Target Stability (DARTS) technique with Mass Spectrometry analysis we identified protein phosphatase 2A (PP2A) as a top ATG-bound protein in cultured renal cells, and this was further confirmed by western blot, computational docking, and surface plasmon resonance assay. In addition, ATG enhanced the activity of PP2A in cultured podocytes and in diabetic glomeruli, resulting in dephosphorylation of p65 NF-κB. Studying the PP2A interacting proteins in podocytes by Mass Spectrometry identified Drebrin-1 (DBN1) as a F- actin interacting protein. Dephosphorylation of DBN1 at T335 by PP2A in podocytes resulted in increased cell adhesion and decreased migration. Importantly, podocyte-specific deletion of Pp2a in mice led to aggravated diabetes-induced podocyte and glomerular injury and the loss of efficacy in ATG-mediated renoprotection. In addition, we found that PPP2R2B, a regulatory subunit of PP2A, expresses uniquely in podocytes in human glomeruli. Its expression is downregulated in the glomeruli of human DKD. In human podocytes, the knockdown of PPP2R2B reduced PP2A activity and expression of PPP2R2B is suppressed by high glucose. Phenome Wide Association Scan identified several missense variants in the human PPP2R2B gene which were associated with either worse or better renal outcomes. These data support a critical role of PP2A in human kidney disease. Based on these observations, we hypothesized that PP2A plays a key role in podocyte biology and pathogenesis of DKD. To test our hypothesis, we will determine the regulation and function of PPP2R2B in podocyte. We will also study the role of PPP2R2B in the regulation of PP2A activity, subcellular localization, and podocyte function in vitro under diabetic conditions and determine whether induction of PPP2R2B or its diseased variant expression in podocytes affects podocyte injury and DKD progression in diabetic mice. We will also determine the downstream signaling of PP2A in podocytes by focusing on the role of DBN1. We will study how PP2A affects podocyte function through regulating DBN1 phosphorylation. The role of DBN1 and its phosphorylation will be also studied in diabetic mice with DKD using transgenic approach. These studies will help us to reveal new insights in the podocyte biology and the pathogenesis of DKD and identify potential new therapy for DKD.

标题：PP2A在足细胞生物学和糖尿病肾脏疾病中的作用 概括 糖尿病肾病（DKD）仍然是慢性肾脏疾病的主要原因，治疗方案有限。 亚氏素（ATG）是源自果汁果的提取物，这是一种传统的中草药 补救措施授予 数据表明，仅ATG给药是衰减小鼠蛋白尿和足细胞损伤的选举权 1型和2型糖尿病的模型。 小鼠表明，受ATG治疗影响的主要途径是细胞和Hondhesion 改善细胞粘附在培养的人足细胞中的迁移。 具有质谱分析的响应式靶稳定性（DARTS）技术 磷酸酶2a（PP2A）作为培养的肾细胞中ATG结合的最高蛋白，这是由此证实的 蛋白质印迹，计算对接和表面等离子体共振测定。 pp2a在培养的足细胞和糖尿病肾小球中的活性，导致p65 NF-κB的去磷酸化。 通过将DREBRIN-1（DBN1）鉴定为F- 肌动蛋白相互作用的蛋白质。 粘附和减少的迁移。 糖尿病诱导的足细胞和肾小球损伤以及ATG介导的肾脏效果丧失 此外，我们发现PP2A的常规亚基PPP2R2B在人类的足细胞中无唯一的表达 肾小球的表达在人dkd的肾小球中被下调。 PPP2R2B的PP2A活性降低，PPP2R2B的表达被高葡萄糖抑制 关联扫描确定了人类PPP2R2B基因中的严重程度变异的变体与 这些数据更糟或更好。 基于观察结果，我们假设PP2A在足细胞和发病机理中扮演关键的rolete 为了测试我们的假设，我们将确定PPP2R2B的调节和功能 还研究PPP2R2B在PP2A活性，亚细胞定位和足细胞功能的调节中的作用 在糖尿病条件下体外，并确定PPP2R2B的诱导还是其患病的变异表达是 在足细胞中，糖尿病小鼠的足细胞损伤和DKD进展。 PP2A的下游信号通过关注DBN1的作用。 通过调节DBN1磷酸化的足细胞功能。 还使用转基因方法在糖尿病小鼠中进行了研究。 对足细胞生物学的见解和DKD的发病机理，并确定了DKDD的潜在新疗法。

项目成果

A central role for mesangial cells in the initiation of diabetic nephropathy.

系膜细胞在糖尿病肾病的发生中起核心作用。

• DOI: 10.1016/j.kint.2023.03.033
• 发表时间: 2023
• 期刊: Kidney international
• 影响因子: 19.6
• 作者: Fang,Zhengying;Lee,Kyung;He,JohnCijiang
• 通讯作者: He,JohnCijiang