青少年肾结核是一种遗传性肾纤毛病，伴有囊性肾病、肾纤维化和儿童和年轻人的终末期肾衰竭。编码 nephrocystin-1 蛋白的 NPHP1 基因突变已被确定为最常见的致病基因，并导致肾髓质囊肿的形成。青少年肾结核的分子发病机制仍然难以捉摸，甚至到今天也没有有效的药物来预防终末期肾衰竭。目前还没有可用的人类细胞模型。在这里，我们报告了第一个青少年肾结核疾病模型，使用患者来源和基因编辑的人类诱导多能干细胞（hiPSC）和源自这些 hiPSC 的肾脏类器官。我们从患者来源的 hiPSC 中建立了 NPHP1 过表达的 hiPSC，并从健康供体 hiPSC 中建立了 NPHP1 缺陷的 hiPSC。比较这一系列 hiPSC，我们发现与 hiPSC 中 NPHP1 缺陷相关的初级纤毛异常。由缺乏 NPHP1 的 hiPSC 产生的肾脏类器官在持续旋转的悬浮培养中经常形成肾囊肿。 NPHP1 的过度表达可以挽救患者来源的肾脏类器官中形成的囊肿。对这些肾脏类器官的转录组分析表明，NPHP1 的缺失导致上皮细胞中与初级纤毛相关的基因表达降低，而与细胞周期相关的基因表达升高。这些发现表明，NPHP1 缺失引起的初级纤毛异常与肾囊肿形成过程中的异常增殖特征之间存在关系。这些发现表明，基于 hiPSC 的青少年肾结核系统疾病模型有助于阐明分子发病机制和开发新疗法。版权所有 © 2024 Arai、Ito、Shimizu、Shimoda、Song、Matsuo-Takasaki、Hayata 和 Hayashi。

Juvenile nephronophthisis is an inherited renal ciliopathy with cystic kidney disease, renal fibrosis, and end-stage renal failure in children and young adults. Mutations in the NPHP1 gene encoding nephrocystin-1 protein have been identified as the most frequently responsible gene and cause the formation of cysts in the renal medulla. The molecular pathogenesis of juvenile nephronophthisis remains elusive, and no effective medicines to prevent end-stage renal failure exist even today. No human cellular models have been available yet. Here, we report a first disease model of juvenile nephronophthisis using patient-derived and gene-edited human induced pluripotent stem cells (hiPSCs) and kidney organoids derived from these hiPSCs. We established NPHP1-overexpressing hiPSCs from patient-derived hiPSCs and NPHP1-deficient hiPSCs from healthy donor hiPSCs. Comparing these series of hiPSCs, we found abnormalities in primary cilia associated with NPHP1 deficiency in hiPSCs. Kidney organoids generated from the hiPSCs lacking NPHP1 formed renal cysts frequently in suspension culture with constant rotation. This cyst formation in patient-derived kidney organoids was rescued by overexpression of NPHP1. Transcriptome analysis on these kidney organoids revealed that loss of NPHP1 caused lower expression of genes related to primary cilia in epithelial cells and higher expression of genes related to the cell cycle. These findings suggested the relationship between abnormality in primary cilia induced by NPHP1 loss and abnormal proliferative characteristics in the formation of renal cysts. These findings demonstrated that hiPSC-based systematic disease modeling of juvenile nephronophthisis contributed to elucidating the molecular pathogenesis and developing new therapies.Copyright © 2024 Arai, Ito, Shimizu, Shimoda, Song, Matsuo-Takasaki, Hayata and Hayashi.