Regulation of glomerular matrix proteins via podocytic and endothelial-cell derived microRNAs

通过足细胞和内皮细胞衍生的 microRNA 调节肾小球基质蛋白

• 批准号: 407053501
• 负责人: Professorin Dr. Janina Müller-Deile
• 金额: --
• 依托单位: Medizinische Klinik 4 - Nephrologie und Hypertensiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/407053501?language=en
• 关键词: Regulation; glomerular; matrix; proteins; via

Proteinuria is a hallmark of glomerular disease. We identified nephronectin (NPNT) as an extracellular matrix protein important for the glomerular filtration barrier. Podocyte microRNA-378a-3p (miR-378a-3p) and endothelial miR-192 regulate podocyte NPNT expression and are upregulated in membranous glomerulonephritis. Overexpression of miR-378a-3p or miR-192 as well as knockdown of npnt caused a phenotype with proteinuria, edema, podocyte effacement and widening of the glomerular basement membrane. Furthermore, we want to analyze the regulation of glomerular matrix proteins via podocyte and endothelial cell derived miRs.We will investigate the glomerular function of npnt and generate podocyte specific npnt knockout mice that will be analyzed regarding spontaneous or stress related development of glomerular damage. Using different Npnt constructs with and without the specific miR binding site we will solve the question to what degree the glomerular changes after miR-378a-3p overexpression are due to npnt knockdown or other miR targets.We will use fluorescent dextrans with different molecular weights to characterize the increase of glomerular permeability after npnt knockdown. To identify other matrix associated targets that are post-transcriptionally controlled by miR-378a-3p and miR-192, we will perform comparative microarrays after miR transfection of human podocytes and glomerular endothelial cells. Finally, we will investigate the therapeutic potential of antagomirs of miR-378a-3p and miR-192 to analyze if inhibition of these miRs has a protective effect.

蛋白尿是肾小球疾病的一个标志。我们确定肾连接素（NPNT）是一种对肾小球滤过屏障很重要的细胞外基质蛋白。足细胞 microRNA-378a-3p (miR-378a-3p) 和内皮 miR-192 调节足细胞 NPNT 表达，并在膜性肾小球肾炎中上调。 miR-378a-3p 或 miR-192 的过度表达以及 npnt 的敲低导致蛋白尿、水肿、足细胞消失和肾小球基底膜增宽的表型。此外，我们希望通过足细胞和内皮细胞衍生的 miR 来分析肾小球基质蛋白的调节。我们将研究 npnt 的肾小球功能，并生成足细胞特异性 npnt 敲除小鼠，对小鼠肾小球损伤的自发或应激相关发展进行分析。使用具有和不具有特定miR结合位点的不同Npnt构建体，我们将解决miR-378a-3p过表达后肾小球变化在多大程度上是由于npnt敲低或其他miR靶标引起的问题。我们将使用不同分子量的荧光葡聚糖来表征npnt敲低后肾小球通透性的增加。为了鉴定由 miR-378a-3p 和 miR-192 转录后控制的其他基质相关靶标，我们将在人足细胞和肾小球内皮细胞转染 miR 后进行比较微阵列。最后，我们将研究 miR-378a-3p 和 miR-192 的 antagomir 的治疗潜力，以分析抑制这些 miR 是否具有保护作用。

项目成果

Circulating factors cause proteinuria in parabiotic zebrafish.

• DOI: 10.1016/j.kint.2019.02.013
• 发表时间: 2019-08
• 期刊: Kidney international
• 影响因子: 19.6
• 作者: J. Müller-Deile;H. Schenk;P. Schroder;K. Schulze;P. Bolaños-Palmieri;F. Siegerist;N. Endlich;H. Haller;M. Schiffer

Identification of cell and disease specific microRNAs in glomerular pathologies

• DOI: 10.1111/jcmm.14270
• 发表时间: 2019-06-01
• 期刊: JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
• 影响因子: 5.3
• 作者: Mueller-Deile, Janina;Dannenberg, Jan;Schiffer, Mario
• 通讯作者: Schiffer, Mario