The identification of key molecules and pathways in the crosstalk of calcium oxalate-treated TCMK-1 cells and macrophage via exosomes

• Published in: Scientific reports 2024-09, Vol.14 (1), p.20949-14, Article 20949
• Main Authors: Sun, Yushi, Li, Bojun, Zhou, Xiangjun, Rao, Ting, Cheng, Fan
• Format: Article
• Language: English
• Subjects: 631/1647/514; 631/250; 692/4017; 692/4022; 692/4025; 692/53; Animals; Biomarkers in kidney stones; Calcium; Calcium oxalate; Calcium Oxalate - metabolism; Calculi; Cell interactions; Cell Line; Crystals; Epithelial cells; Exosomes; Exosomes - metabolism; Gene expression; Gene Expression Regulation - drug effects; Humanities and Social Sciences; Humans; Kidney Calculi - genetics; Kidney Calculi - metabolism; Kidney stones; Kidneys; Macrophage; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Mice; MicroRNAs - genetics; MicroRNAs - metabolism; MiRNA; multidisciplinary; Nephrolithiasis; Next-generation sequencing; Oxalic acid; RAW 264.7 Cells; Science; Science (multidisciplinary); Signal Transduction - drug effects; Transcriptomics of kidney stones
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-71755-y

The interplay between crystals and epithelial cells forms the cornerstone of kidney stone development, communication between epithelial cells and macrophages emerging as a pivotal role in this process. We conducted next-generation sequencing on the secreted exosomes of TCMK-1 cells treated with calcium oxalate monohydrate (OX_EXO) or controls (NC_EXO), and on the macrophage cell line RAW264.7 stimulated with OX_EXO or NC_EXO, followed by validation of differentially expressed target proteins and miRNAs through Western blot and PCR. UPSET plots were employed to identify genes co-targeted by exosomal miRNAs. Various bioinformatic analyses were employed to predict potential mechanisms of the dysregulated genes. We integrated sequencing data from the GEO database, and validated findings using clinical patient urine and kidney tissues. We identified 665 differentially expressed exosomal miRNAs between OX_EXO and NC_EXO. Among the top 10 down-regulated miRNAs, the most targeted genes were AAK1 and NUFIP2, whereas PLCB1 was significantly targeted among the top 10 up-regulated miRNAs. In clinical specimens, we confirmed the differential expressions of five homologous miRNAs, as well as CNOT3, CNCNA1C, APEX1, and TMEM199. In conclusion, treatment of TCMK-1 cells with calcium oxalate significantly alerted the expression profile of exosomal miRNAs, subsequently influencing gene expression in macrophages, thereby modulating the processes of kidney stone formation.