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Merck

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

Cell stem cell (2022-07-09)
Tracy Tran, Cheng Jack Song, Trang Nguyen, Shun-Yang Cheng, Jill A McMahon, Rui Yang, Qiuyu Guo, Balint Der, Nils O Lindström, Daniel C-H Lin, Andrew P McMahon
RÉSUMÉ

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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Description du produit

Roche
Cocktail d'inhibiteurs de protéases sans EDTA Mini cOmplete, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial
Sigma-Aldrich
CHIR99021, ≥98% (HPLC)
Sigma-Aldrich
Anti-SLC12A1 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-SLC3A1 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution