Analysis of high glucose injury using human induced pluripotent stem cell-derived kidney organoids

BACKGROUND: Diabetic kidney disease (DKD) is one of the most pervasive complications of diabetes worldwide. However, the pathogenesis of DKD remains poorly understood, due to limitations of the models. The hPSC-derived kidney organoids may offer a new possibility to solve the problem. METHODS: We generated human pluripotent stem cells (hPSCs) derived kidney organoids to model DKD injury by glucose intervention for 24 and 72 h, respectively. RT-qPCR was used to assess gene expression, while immunofluorescence was performed to evaluate protein expression. PAS staining was applied to examine organoid morphology, and Sirius Red staining was used to assess fibrosis. RESULTS: Firstly, qPCR results showed that glucose and lipid metabolism-related genes such as LDH, HK2, SGLT2, PLIN2, PPARA, PGC1A, and HSL mRNA expression were upregulated after glucose intervention. Secondly, qPCR and immunofluorescence staining results revealed that the expression of pro-inflammatory cytokines IL6, IL1B, TNFA, VCAM1 and IL-10 were increased, which suggested kidney organoids possess inflammatory responses in high glucose environments. Thirdly, KIM1, a kidney injury maker was upregulated after glucose intervention, and increased apoptosis cells in kidney organoids were confirmed by TUNEL assay. Finally, qPCR and immunofluorescence staining results revealed that the expression of fibrosis-related molecules TGF-β1 and COL4 were increased. CONCLUSION: In general, diabetic kidney disease organoid models provide a valuable model for studying the onset, progression, and injury of DKD. CLINICAL TRIAL NUMBER: Not applicable.