Abstract
Background: Exosomes, as a mediator of cell-to-cell communication has regulatory effects on the genetic and epigenetic processes in the recipient cells. Although the ability of human induced pluripotent stem cells (hiPSCs) derived exosomes to participate in tissue repair is being increasingly recognized, the information inside the hiPSC-exosomes especially protein content is not completely identified yet. Method: Exosomes derived from human fibroblast cell line IMR90 and IMR90 derived hiPSC cell line iPS (IMR90)-4 were collected using differential ultracentrifugation, then the exosomal proteins were identified by mass spectrometry analysis. The proteomics results were confirmed by western blot. To validate the role hiPSC-exosome in epigenetic processes, the H9C2 cells were treated with both types of exosomes and the histone epigenetic state were analyzed. Results: Gene ontology (GO) analysis of the exosome proteome showed that the hiPSC-exosome protein content had statistically significant enrichment in epigenetic modification such as acetylation, phosphorylation and methylation. The proteomic analysis indentified 6 epigenetic enzymes (UBE2N, DNMT3B, AURKB, CARM1, HDAC2 and HAT1) which only expressed in hiPSC-exosomes whereas 2 enzymes (PRMT5 and PRMT1) showed increased level in hiPSC-exosomes. The differential expressions of HDAC2 and PRMT5 were validated by western blot. Furthermore, H9C2 cell treated with hiPSC-exosomes showed a significant decrease in pan-acetylation level of H4 compared with IMR90-exosomes. Conclusion: In a global analysis presented here, hiPSC-exosomal proteins have been found to be associated with mammalian epigenetic inheritance, which provide important information for the exosome-based therapy.
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