APOL1 in Diabetic Podocytopathy

糖尿病足细胞病中的 APOL1

基本信息

批准号：
9767114
负责人：
PRAVIN C SINGHAL
金额：
$ 49.98万
依托单位：
FEINSTEIN INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-20 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9767114
关键词：
APOL1 gene Agonist Apolipoproteins Apoptosis Biological Assay Cell Death Cell physiology Cells Chronic Kidney Failure Development Diabetic Nephropathy Diabetic mouse Differentiation Antigens Diffuse Down-Regulation Embryonic Development End stage renal failure Epithelial Cells Evaluation Feedback Glucose Goals Human In Vitro Injury Kidney Lesion Lipids Maintenance Mediating Mesenchymal MicroRNAs Minor Molecular Structure Multiprotein Complexes Oxidative Stress Parietal Play Population Prevention Primates Proteinuria Regulation Role Structure Therapeutic Tubular formation Up-Regulation Vitamin D3 Receptor WT1 gene Western Blotting base claudin-1 protein diabetic diabetic patient genetic epidemiology glomerular filtration glomerulosclerosis in vivo insight kidney cell molecular marker molecular phenotype nephrin overexpression podocalyxin podocyte preservation prevent risk variant slit diaphragm tool transcription factor

项目摘要

Diabetic patients develop podocytopathy due to the loss of podocyte function caused by dedifferentiation and cell death (apoptosis), or both. In injury mode, podocytes dedifferentiate- loss of their molecular markers (WT1 and nephrin) and gain of parietal epithelial cell markers such as PAX2 and claudin 1. High glucose milieu is known to enhance dedifferentiation and apoptosis of podocytes both in vitro and in vivo studies. Kidney cells including podocytes and tubular cells express apolipoprotein (APO) L1, however, the role of APOL1 expression in kidney cell function is not clear. The objective of the current proposal is to determine the role of APOL1 in the maintenance of podocyte integrity in high glucose milieu. The long-term goal is to utilize the optimized expression of APOL1 by podocytes as a therapeutic strategy to prevent and/or slow down the progression of diabetic podocytopathy. In preliminary studies, high glucose enhanced microRNA (miR)193a but down regulated APOL1 expression in podocytes; interestingly, miR193a and APOL1 displayed inverse feedback relationship. Since miR193a negatively regulates WT1, a transcription factor for podocyte differentiating markers, miR193a-APOL1 axis would also participate in the regulation of differentiation status in podocytes. In preliminary studies, both down regulation of miR193a and upregulation of APOL1 provided protection against high glucose mediated dedifferentiation/apoptosis in podocytes; furthermore, vitamin D receptor agonist (VDA), which down regulated miR193a and upregulated APOL1 expression, preserved podocytes integrity in high glucose milieu. Based on these findings, we hypothesize that APOL1 acts as an important component of miR193a-APOL1 axis to preserve podocyte integrity. High glucose compromises podocyte integrity through upregulation of miR193a and down regulation of APOL1, whereas, optimization of APOL1 expression would prevent and/or slow down the progression of diabetic podocytopathy. To validate our hypotheses, we will carry out the following specific aims:  Evaluation of the role of the miR193a-APOL1 axis on PDs molecular phenotype and structural integrity  Determination whether miR193a expression would accelerate but APOL1 expression would inhibit high glucose-induced PD injury both in vitro and in vivo  Examination of the role of VDA in prevention and/or slow down the progression of diabetic podocytopathy with or without APOL1 expression This proposal provides mechanistic insight into the development of diabetic podocytopathy. We feel strongly that optimization of APOL1 expression in PDs could be used as a therapeutic strategy to preserve PDs molecular phenotype and structural integrity in the diabetic milieu.

糖尿病患者由于去分化和细胞死亡（凋亡）或两者兼而有之。在损伤模式下，足细胞去分化的分子标记损失（WT1）和肾素）和顶上皮细胞标记（例如Pax2和claudin 1）的获得。高葡萄糖环境是已知可以增强体外和体内研究的足细胞的去分化和凋亡。肾细胞但是，包括足细胞和管状细胞表达载脂蛋白（APO）L1，但是，Apol1的作用肾细胞功能中的表达不清楚。当前建议的目的是确定在高葡萄糖环境中维持足细胞完整性的apol1。长期目标是利用通过足细胞优化apol1的表达，作为预防和/或放慢速度的治疗策略糖尿病足细胞病的进展。在初步研究中，高葡萄糖增强的microRNA（MIR）193a 但在足细胞中调节的apol1表达；有趣的是，mir193a和apol1表现为逆反馈关系。由于miR193a负调节WT1，因此是足细胞的转录因子区分标记，miR193a-apol1轴也将参与分化状态的调节在足细胞中。在初步研究中，MiR193a的下调和APOL1的上调提供了保护足细胞中高葡萄糖介导的去分化/凋亡；此外，维生素d 受体激动剂（VDA）下调了miR193a并更新Apol1表达式，保留了高葡萄糖环境中的足细胞完整性。基于这些发现，我们假设Apol1充当 miR193a-apol1轴的重要组成部分，以保留足细胞完整性。高葡萄糖妥协通过上调mir193a和Apol1的下调，足细胞完整性，而优化 APOL1表达将预防和/或减慢糖尿病足病毒的进展。验证我们的假设，我们将执行以下具体目标： 评估miR193a-apol1轴对PDS分子表型和结构完整性的作用 确定miR193a表达是否会加速，但APOL1表达会抑制高葡萄糖诱导的PD损伤在体外和体内都 检查VDA在预防和/或减慢糖尿病进展中的作用有或没有APOL1表达的足细胞病该提案为糖尿病足细胞病的发展提供了机械洞察力。我们感到坚强 PDS中APOL1表达的优化可以用作保存PDS的治疗策略糖尿病环境中的分子表型和结构完整性。