Unveiling the functional heterogeneity of endogenous tissue extracellular vesicles in skeletal muscle through multi-omics

Abstract

Extracellular vesicles (EVs) have emerged as promising tools for the development of disease biomarkers and therapeutics because they can transfer various bioactive cargos between cells in vivo. A better understanding of the heterogeneous properties of EVs in vivo may provide insights into their biological roles and clinical translation potential. As a proof-of-concept, we report that different EV subpopulations from skeletal muscle tissues have distinct composition signatures and diverse biological effects on recipient cells. Multiple cell types (e.g., myoblasts and endothelial cells (ECs)) can contribute to the pool of muscle tissue-derived EVs, and large EVs (L-EVs) are enriched with proteins related to metabolic regulation, whereas small EVs (S-EVs) are enriched with original muscle cell-specific proteins related to muscle function regulation. Compared with L-EVs, S-EVs exhibited abundant surface proteins and higher cell uptake rates. Moreover, L-EVs and S-EVs can induce diverse changes in global gene expression, metabolic patterns and some cellular behaviors (e.g., proliferation and differentiation) in recipient cells. These results suggest that different EV subpopulations might control tissue hemostasis in a coordinated manner and suggest the importance of reconsidering their favorable role in future applications (e.g., S-EVs for biomarker discovery and L-EVs for metabolic intervention). This study highlights the functional heterogeneity of tissue-derived EVs in vivo, and the selection of an ideal EV subset on the basis of its specific biological properties may be a promising strategy for developing more precise biomarkers or tailored EV therapies for regenerative medicine. Graphic abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=60 SRC="FIGDIR/small/624461v1_ufig1.gif" ALT="Figure 1"> View larger version (23K): org.highwire.dtl.DTLVardef@6a9a0forg.highwire.dtl.DTLVardef@a86cbforg.highwire.dtl.DTLVardef@17b8680org.highwire.dtl.DTLVardef@bb718d_HPS_FORMAT_FIGEXP M_FIG C_FIG SignificanceExtracellular vesicles (EVs) are emerging as important tools for diagnostics, therapeutics, and prognostics in various diseases. Understanding the inherent heterogeneity of EVs is crucial, as distinct subpopulations function differently. While extensive research focuses on EVs derived from cell supernatants, endogenous tissue EVs can more accurately reflect the pathophysiological characteristics of their originating cells. Here, we propose different tissue-EV subpopulations coordinately regulate tissue homeostasis. Small EVs with tissue-specific signatures show promise for biomarker development, while large EVs with metabolic signatures are suitable for metabolic interventions. Additionally, small EVs with enhanced surface proteins are ideal for targeted delivery. This work highlights the importance of selecting tissue-EV subpopulations based on their unique properties for developing precise biomarkers and tailored therapies in regenerative medicine.