목적: Permanent hearing loss primarily results from the inability of the mammalian cochlea to replace lost inner ear hair cells. However, neonatal mice exhibit a unique capacity: isolated cochlear floor cells can efficiently proliferate in vitro and form organoids that harbor new hair cells and supporting cell populations. In this study, we isolated extracellular vesicles (EVs) from organoids, analyzed the miRNAs derived from them, and compared those with publicly available dataset to identify gene regulatory elements that coordinate inner ear proliferation and regeneration. 방법:We utilized cochlear floor cells from postnatal day two mice and optimized the culture conditions to efficiently grow organoids that exhibit progenitor properties. Next, we isolated EVs from the culture media of organoids in their proliferative and differentiation stage. We analyzed miRNAs contained in these EVs to identify potential regulators that drive or modulate organoid generation. miRNA sequencing data from organoid EVs were then compared with mRNA-sequencing data and single cell sequencing data from publicly available dataset. 결과:We identified 138 mature miRNAs in organoids EVs. A total of 91 miRNAs differed more than 2-fold between these groups, with 35 miRNAs (6 upregulated and 29 downregulated in organoid EVs) exhibiting statistically significant differences. Target gene prediction and pathway analysis resulted in 7 miRNAs and 195 targets genes related with the organoid differentiation. Further target gene prediction, protein-protein network, and trajectory analysis revealed 18 genes (9 upregulated and 9 downregulated) related with organoid differentiation. In GO analysis (biological process), these target genes were associated with pathways related to epithelial cell differentiation, regulation of gliogenesis, glial cell proliferation, regulation of neurogenesis, small GTPase mediated signal transduction, Ras protein signal transduction, and cellular response to organonitrogen compound. This indicates that the miRNAs in organoid- derived EVs may impact processes associated with cell differentiation and the generation of inner ear cell types. 결론:Our study comprehensively inventoried miRNAs contained in EVs released by growing inner ear organoids. Our differential miRNA expression analysis provides insight into regulatory mechanisms that promote cochlear floor cell proliferation and organoid formation, which could be leveraged in miRNA-based therapeutic approaches.