Research Paper Advance Articles
Exosomes released from senescent cells and circulatory exosomes isolated from human plasma reveal aging-associated proteomic and lipid signatures
Figure 1. Multi-omics workflow for aging/senescence exosome biomarker discovery. (A) Two different sample types, primary human lung fibroblasts and human plasma, were investigated. Three senescence stimuli, Irradiation; IR, Doxorubicin; doxo, and Antimycin A-induced mitochondrial dysfunction-associated senescence; MiDAS, were used to generate senescent primary human lung fibroblasts (n=4, each condition). Quiescent cells (Qui) or DMSO-treated fibroblasts were used as control. Plasma from young (20-26 years old) and older (65-74 years old) individuals was used (n=5 each). Exosomes were enriched by sequential size-exclusion chromatography and ultrafiltration (SEC/UF). Immunoblotting and particle size analysis was conducted to confirm enriched, high-quality, and intact exosomes. Three different protein spectral libraries; DDA, directDIA, and hybrid, were generated. Multicomponent Senescence/Aging Biomarkers were identified by high-throughput quantitative exosome multi-omics (proteomics, lipidomics, and miRNA) analysis.