Müller-Deile Janina, Schenk Heiko, Niggemann Philipp, Bolaños-Palmieri Patricia, Teng Beina, Higgs Alysha, Staggs Lynne, Haller Hermann, Schroder Patricia, Schiffer Mario
Department of Nephrology, Hannover Medical School, Hannover 30625, Germany.
Department of Nephrology and Hypertension, University of Erlangen-Nurnberg, Erlangen 91054, Germany.
Biol Open. 2019 Mar 4;8(3):bio040253. doi: 10.1242/bio.040253.
Different glomerular diseases that affect podocyte homeostasis can clinically present as nephrotic syndrome with massive proteinuria, hypoalbuminemia, hyperlipidemia and edema. Up to now, no drugs that specifically target the actin cytoskeleton of podocytes are on the market and model systems for library screenings to develop anti-proteinuric drugs are of high interest. We developed a standardized proteinuria model in zebrafish using puromycin aminonucleoside (PAN) via treatment in the fish water to allow for further drug testing to develop anti-proteinuric drugs for the treatment of glomerular diseases. We noticed that fish that carry the nacre-mutation show a significantly higher susceptibility for the disruption of the glomerular filtration barrier following PAN treatment, which results in a more pronounced proteinuria phenotype. Nacre zebrafish inherit a mutation yielding a truncated version of microphthalmia-associated transcription factor/melanogenesis associated transcription factor (mitf). We hypothesized that the nacre mutation may lead to reduced formin expression and defects in cytoskeletal rearrangement. Based on the observations in zebrafish, we carried out a PAN treatment on cultured human podocytes after knockdown with MITF siRNA causing a rearrangement of the actin cytoskeleton.
不同影响足细胞内环境稳定的肾小球疾病在临床上可表现为肾病综合征，伴有大量蛋白尿、低蛋白血症、高脂血症和水肿。到目前为止，市场上还没有专门针对足细胞肌动蛋白细胞骨架的药物，用于开发抗蛋白尿药物的文库筛选模型系统备受关注。我们通过在鱼水中使用嘌呤霉素氨基核苷（PAN）在斑马鱼中建立了标准化的蛋白尿模型，以便进行进一步的药物测试，开发用于治疗肾小球疾病的抗蛋白尿药物。我们注意到携带珍珠母突变的鱼在接受PAN治疗后，肾小球滤过屏障破坏的易感性显著更高，这导致更明显的蛋白尿表型。珍珠母斑马鱼继承了一个突变，产生了小眼相关转录因子/黑色素生成相关转录因子（mitf）的截短版本。我们假设珍珠母突变可能导致formin表达减少和细胞骨架重排缺陷。基于在斑马鱼中的观察结果，我们在用MITF siRNA敲低导致肌动蛋白细胞骨架重排的培养人足细胞上进行了PAN处理。
