Tabatabaeifar Mansoureh, Wlodkowski Tanja, Simic Ivana, Denc Helga, Mollet Geraldine, Weber Stefanie, Moyers John Julius, Brühl Barbara, Randles Michael Joseph, Lennon Rachel, Antignac Corinne, Schaefer Franz
Division of Pediatric Nephrology, Center for Pediatrics and Adolescent Medicine, University of Heidelberg, Heidelberg, Germany.
INSERM, U1163, Imagine Institute, Laboratory of Hereditary Kidney Diseases, Paris, France.
PLoS One. 2017 Oct 19;12(10):e0186574. doi: 10.1371/journal.pone.0186574. eCollection 2017.
Mutations in the NPHS2 gene, encoding podocin, cause hereditary nephrotic syndrome. The most common podocin mutation, R138Q, is associated with early disease onset and rapid progression to end-stage renal disease. Knock-in mice carrying a R140Q mutation, the mouse analogue of human R138Q, show developmental arrest of podocytes and lethal renal failure at neonatal age. Here we created a conditional podocin knock-in model named NPHS2 R140Q/-, using a tamoxifen-inducible Cre recombinase, which permits to study the effects of the mutation in postnatal life. Within the first week of R140Q hemizygosity induction the animals developed proteinuria, which peaked after 4-5 weeks. Subsequently the animals developed progressive renal failure, with a median survival time of 12 (95% CI: 11-13) weeks. Foot process fusion was observed within one week, progressing to severe and global effacement in the course of the disease. The number of podocytes per glomerulus gradually diminished to 18% compared to healthy controls 12-16 weeks after induction. The fraction of segmentally sclerosed glomeruli was 25%, 85% and 97% at 2, 4 and 8 weeks, respectively. Severe tubulointerstitial fibrosis was present at later disease stage and was correlated quantitatively with the level of proteinuria at early disease stages. While R140Q podocin mRNA expression was elevated, protein abundance was reduced by more than 50% within one week following induction. Whereas miRNA21 expression persistently increased during the first 4 weeks, miRNA-193a expression peaked 2 weeks after induction. In conclusion, the inducible R140Q-podocin mouse model is an auspicious model of the most common genetic cause of human nephrotic syndrome, with a spontaneous disease course strongly reminiscent of the human disorder. This model constitutes a valuable tool to test the efficacy of novel pharmacological interventions aimed to improve podocyte function and viability and attenuate proteinuria, glomerulosclerosis and progressive renal failure.
编码足突蛋白的NPHS2基因突变会导致遗传性肾病综合征。最常见的足突蛋白突变R138Q与疾病早期发作及快速进展至终末期肾病有关。携带R140Q突变(人类R138Q的小鼠类似物)的基因敲入小鼠在新生儿期显示足细胞发育停滞和致命性肾衰竭。在此,我们利用他莫昔芬诱导型Cre重组酶创建了一个名为NPHS2 R140Q/-的条件性足突蛋白基因敲入模型,该模型允许在出生后研究该突变的影响。在诱导R140Q半合子后的第一周内,动物出现蛋白尿,在4-5周后达到峰值。随后,动物出现进行性肾衰竭,中位生存时间为12周(95%置信区间:11-13周)。在一周内观察到足突融合,在疾病过程中进展为严重且全面的足突消失。与诱导后12-16周的健康对照相比,每个肾小球的足细胞数量逐渐减少至18%。节段性硬化肾小球的比例在2周、4周和8周时分别为25%、85%和97%。在疾病后期出现严重的肾小管间质纤维化,并且在疾病早期阶段与蛋白尿水平呈定量相关。虽然R140Q足突蛋白mRNA表达升高,但在诱导后一周内蛋白质丰度降低了50%以上。在最初4周内,miRNA21表达持续增加,而miRNA-193a表达在诱导后2周达到峰值。总之,诱导型R140Q-足突蛋白小鼠模型是人类肾病综合征最常见遗传病因的一个良好模型,其自发病程强烈让人联想到人类疾病。该模型是一个有价值的工具,可用于测试旨在改善足细胞功能和活力以及减轻蛋白尿、肾小球硬化和进行性肾衰竭的新型药物干预的疗效。