Abstract: The development of kidney organoids holds significant implications for both fundamental research and clinical applications in renal diseases. Despite remarkable advances in kidney organoid research in recent years, several challenges still exist. Through systematic screening of small molecules and recombinant proteins, we identified critical roles of retinoic acid (RA), glial cell line-derived neurotrophic factor (GDNF), and fibroblast growth factor 20 (FGF20) in enabling long-term in vitro expansion of kidney organoids. Immunofluorescence staining and single-cell RNA sequencing analyses confirmed the concurrent differentiation of proximal tubule and collecting duct lineages within these organoids. Leveraging this culture system, we generated polycystic kidney disease (PKD) organoids carrying PKD1 or PKD2 mutations using renal tissues from autosomal dominant polycystic kidney disease (ADPKD) patients. Single-cell transcriptomic profiling validated that PKD organoids recapitulated key molecular features of native PKD tissues. Furthermore, comparative analysis revealed cellular heterogeneity between PKD1- and PKD2-mutant organoids, underscoring their capacity to mirror the phenotypic diversity of patient-derived tissues. Based on these findings, we established a high-throughput drug screening platform using cystic structure diameter as a quantifiable endpoint. Our study presents a groundbreaking adult kidney-derived organoid culture system capable of sustaining in vitro expansion. By integrating patient-specific tissues, we further developed a robust platform for drug discovery in polycystic kidney disease and identified a promising therapeutic agent. These advancements enrich the foundational understanding of adult renal cell differentiation into organoids, provide a reliable model for studying adult kidney diseases, and demonstrate substantial scientific and clinical translational potential.
Funding Source: the National Natural Science Foundation of China (32122031 to Huili Hu and T2321004 to Huili Hu)
