A scalable organoid model of human autosomal dominant polycystic kidney disease for disease mechanism and drug discovery

• Published in: Cell stem cell Vol. 29; no. 7; pp. 1083 - 1101.e7
• Main Authors: Tran, Tracy, Song, Cheng Jack, Nguyen, Trang, Cheng, Shun-Yang, McMahon, Jill A., Yang, Rui, Guo, Qiuyu, Der, Balint, Lindström, Nils O., Lin, Daniel C.-H., McMahon, Andrew P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 07-07-2022
• Subjects: Cysts - metabolism; developmental trajectory; disease modeling; Drug Discovery; drug screen; high throughput; Humans; Kidney - metabolism; kidney development; kidney organoid; nephron development; Organoids - metabolism; phenotypic screen; polycystic kidney disease; Polycystic Kidney, Autosomal Dominant - drug therapy; Polycystic Kidney, Autosomal Dominant - genetics; Polycystic Kidney, Autosomal Dominant - metabolism; scRNA-seq; TRPP Cation Channels - genetics; TRPP Cation Channels - metabolism; scRNA-seq; kidney development; developmental trajectory; polycystic kidney disease; disease modeling; phenotypic screen; kidney organoid; drug screen; high throughput; nephron development
• ISSN: 1934-5909; 1875-9777
• DOI: 10.1016/j.stem.2022.06.005

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 genes PKD1 and PKD2 were inactivated by gene editing. PKD1 and PKD2 mutant 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.
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•Scalable human kidney organoid platform with 1–2 nephron-like structures/organoid•Comparative scRNA-seq profiling of in vivo and in vitro kidney programs•PKD1 and PKD2 mutant kidney organoids display efficient cyst formation•Identification of cyst-inhibitory compounds from small-molecule screens
We describe a scalable platform to efficiently generate thousands of comparable human kidney organoids. In this, PKD1 and PKD2 mutant organoids displayed robust cystogenesis. We generated a small-molecule screening workflow that identified compounds inhibiting cyst formation and later growth. The platform will facilitate kidney disease modeling and high-throughput drug screens.