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Distinct non-coding RNA cargo of extracellular vesicles from M1 and M2 human primary macrophages
Paschalia Pantazi1, Toby Clements1, Morten Venø2
1Institute of Reproductive and Developmental Biology, Department of Metabolism, Digestion, and Reproduction, Imperial College London, London, UK.
Abstract:
Macrophages are important antigen presenting cells which can release extracellular vesicles (EVs) carrying functional cargo including non-coding RNAs. Macrophages can be broadly classified into M1 'classical' and M2 'alternatively-activated' macrophages. M1 macrophages have been linked with inflammation-associated pathologies, whereas a switch towards an M2 phenotype indicates resolution of inflammation and tissue regeneration. Here, we provide the first comprehensive analysis of the small RNA cargo of EVs from human M1 and M2 primary macrophages. Using small RNA sequencing, we identified several types of small non-coding RNAs in M1 and M2 macrophage EVs including miRNAs, isomiRs, tRNA fragments, piRNA, snRNA, snoRNA and Y-RNA fragments. Distinct differences were observed between M1 and M2 EVs, with higher relative abundance of miRNAs, and lower abundance of tRNA fragments in M1 compared to M2 EVs. MicroRNA-target enrichment analysis identified several gene targets involved in gene expression and inflammatory signalling pathways. EVs were also enriched in tRNA fragments, primarily originating from the 5' end or the internal region of the full length tRNAs, many of which were differentially abundant in M1 and M2 EVs. Similarly, several other small non-coding RNAs, namely snRNAs, snoRNAs and Y-RNA fragments, were differentially enriched in M1 and M2 EVs; we discuss their putative roles in macrophage EVs. In conclusion, we show that M1 and M2 macrophages release EVs with distinct RNA cargo, which has the potential to contribute to the unique effect of these cell subsets on their microenvironment.
Insights
Macrophages release extracellular vesicles (EVs) with distinct small non-coding RNA cargo. M1 macrophage EVs show more microRNAs, while M2 EVs contain more tRNA fragments, impacting their cellular functions.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Macrophages are key antigen-presenting cells involved in immune responses.
- Macrophages differentiate into M1 (pro-inflammatory) and M2 (anti-inflammatory/regenerative) phenotypes.
- Macrophages release extracellular vesicles (EVs) containing functional non-coding RNAs.
Purpose of the Study:
- To conduct the first comprehensive analysis of small non-coding RNA cargo in EVs from human M1 and M2 macrophages.
- To identify differences in RNA content between M1 and M2 macrophage-derived EVs.
- To explore the potential roles of these distinct RNA cargoes in modulating cellular microenvironments.
Main Methods:
- Isolation and characterization of extracellular vesicles (EVs) from primary human M1 and M2 macrophages.
- Small RNA sequencing to profile the non-coding RNA content of M1 and M2 EVs.
- Bioinformatic analysis, including microRNA-target enrichment and differential abundance analysis.
Main Results:
- Identified diverse small non-coding RNAs (miRNAs, isomiRs, tRNA fragments, piRNAs, snRNAs, snoRNAs, Y-RNA fragments) in macrophage EVs.
- Observed distinct RNA profiles: M1 EVs had higher relative miRNA abundance and lower tRNA fragments compared to M2 EVs.
- Found differential enrichment of various small non-coding RNAs, including tRNA fragments, snRNAs, snoRNAs, and Y-RNA fragments, between M1 and M2 EVs.
Conclusions:
- Human M1 and M2 macrophages release EVs with significantly different small non-coding RNA cargo.
- These distinct RNA profiles suggest a role for macrophage EVs in mediating the unique functions of M1 and M2 subsets.
- The findings provide novel insights into intercellular communication mediated by macrophage-derived EVs in health and disease.
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