Red blood cell-derived extracellular vesicles mediate intercellular communication in ischemic heart failure

Abstract

Extracellular vesicles (EV) mediate intercellular signaling by transferring their cargo to recipient cells. Red blood cell (RBC)-derived EVs constitute a significant proportion of circulating EVs and have been implicated in regulating immune responses. Here, we describe a transgenic mouse model for fluorescent-based mapping of RBC-EV target cells based on the functional transfer of EV-contained Cre-recombinase to target cells. In a murine model of ischemic heart failure, we detect an increase in RBC-EV-targeted cardiomyocytes in the hearts and microglial cells in the brains. Cells targeted by RBC-EVs present an enrichment of genes implicated in cell proliferation and metabolism pathways compared to non-recombined (non-targeted) cells. Cardiomyocytes targeted by RBC-EVs are more likely to demonstrate cellular markers of DNA synthesis and proliferation, suggesting functional significance of EV-mediated signaling. In conclusion,we leverage our mouse model for mapping of RBC-EV targets in murine ischemic heart failure to demonstrate quantitative and qualitative changes in RBC-EV recipients.

Unpacking extracellular vesicles: RNA cargo loading and function

“…However, when EV-producing cells were transfected with vesicular stomatitis virus G (VSVG) plasmids, which incorporate into the EV membrane and promote fusion with recipient cells, mRNA transfer could be confirmed, in two separate studies (Albanese et al, 2021;Somiya and Kuroda, 2022). In vivo, Cre recombination in the reporter system developed by Ridder et al was also dramatically higher in the context of chronic inflammation or myocardial infarction (Das et al, 2019;Ridder et al, 2014). Together these studies demonstrate that whilst functional transfer of mRNA is possible, much more work is needed to understand what specific circumstances it does occur in.…”

“…Stress treatments have also been suggested to increase delivery via EVs, with irradiation of recipient cells shown to increase EV uptake (Mutschelknaus et al., 2016 ), and mRNA delivery increased by multiple different stress treatments (heat shock, hydrogen peroxide, dithiothreitol or serum starvation) (Haimovich et al., 2017 ). Given that in vivo tools developed to report EV‐mediated communication displayed increased reporter signalling during chronic inflammation or myocardial infarction (Das et al., 2019 ; Ridder et al., 2014 ), the healthy functioning of the endolysosomal system of the recipient cell may be critical to cytoplasmic delivery of EV‐RNA. This perhaps provides an explanation to the variable conclusions on RNA transfer that have been seen in other studies, but also indicating that in pathological scenarios where endolysosomal function may be particularly impaired (such as in neurodegenerative disease), delivery could be significantly altered.…”

“…In vivo, Cre recombination in the reporter system developed by Ridder et al. was also dramatically higher in the context of chronic inflammation or myocardial infarction (Das et al., 2019 ; Ridder et al., 2014 ). Together these studies demonstrate that whilst functional transfer of mRNA is possible, much more work is needed to understand what specific circumstances it does occur in.…”

Analysis of Tumor-Derived Exosomes by Nanoscale Flow Cytometry

“…This optimized instrument has allowed for the analysis of EVs obtained from patients' plasma as well as the characterization of exosomes obtained from in lung macrophage supernatants [75,76]. Lately, this equipment has been replaced by the AstriosEQ (Beckman Coulter) which has two FSC detectors optimized for the visualization of EVs between 30 and 200 nm, which is an important leap in the sensitivity of the instruments for the measurement of plasma EVs [77], red blood [78], viruses [64], and cancer EV sorting with satisfactory results [58].…”

Phase 2 of extracellular RNA communication consortium charts next-generation approaches for extracellular RNA research

“…In the field of cardiovascular disease, Das and Van Keuren-Jensen have reported new methods and pipelines for performing long RNAseq (transcriptome) from plasma and EVs with a goal of identifying tissue-specific transcripts ( Rodosthenous et al., 2020 ), and further demonstrated a role for RBC-derived EVs in a mouse model of ischemic heart disease ( Das et al., 2019 ).…”