A scalable organoid model of human autosomal dominant polycystic kidney disease for disease mechanism and drug discovery.
A scalable organoid model of human autosomal dominant polycystic kidney disease for disease mechanism and drug discovery.
复制标题
人类常染色体显性多囊肾病的可扩展类器官模型，用于疾病机制和药物发现。
DOI：
10.1016/j.stem.2022.06.005
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发表时间：
2022
期刊：
影响因子：
23.9
通讯作者：
McMahon,AndrewP
中科院分区：
文献类型：
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作者：
Tran,Tracy;Song,ChengJack;Nguyen,Trang;Cheng,Shun-Yang;McMahon,JillA;Yang,Rui;Guo,Qiuyu;Der,Balint;Lindström,NilsO;Lin,DanielC-H;McMahon,AndrewP
人类多能干细胞衍生的类器官是人类发育和疾病的模型。我们报告了一种改良的人类肾脏类器官系统，该系统产生数千个类似的类器官，每个类器官由1-2个肾单位样结构组成。单细胞转录组学分析和免疫荧光验证强调了模式化的肾单位样结构，其利用与人类肾发生相似的途径，具有不同的形态发生。为了验证该平台用于治疗筛选的可行性，我们通过基因编辑的方法对多囊肾病基因PKD 1和PKD 2进行了失活，并建立了高效、可重复的突变模型。囊性增生可以繁殖几个月厘米大小的囊肿。为了揭示囊肿发生的新情况，在活体成像试验中筛选了247种蛋白激酶抑制剂（PKIs），鉴定了阻断囊肿形成但不阻断整体类器官生长的化合物。类器官平台的扩展和进一步开发将使肾脏疾病建模和高通量药物筛选的能力更广泛。
Human pluripotent stem-cell-derived organoids are models for human development and disease. We report a modified human kidney organoid system that generates thousands of similar organoids, each consisting of 1–2 nephron-like structures. Single-cell transcriptomic profiling and immunofluorescence validation highlighted patterned nephron-like structures utilizing similar pathways, with distinct morphogenesis, to human nephrogenesis. To examine this platform for therapeutic screening, the polycystic kidney disease genesPKD1andPKD2were inactivated by gene editing.PKD1andPKD2mutant models exhibited efficient and reproducible cyst formation. Cystic outgrowths could be propagated for months to centimeter-sized cysts. To shed new light on cystogenesis, 247 protein kinase inhibitors (PKIs) were screened in a live imaging assay identifying compounds blocking cyst formation but not overall organoid growth. Scaling and further development of the organoid platform will enable a broader capability for kidney disease modeling and high-throughput drug screens.