Hypoxic Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Release Large Extracellular Vesicles That Promote a Pro-Inflammatory Phenotype in Macrophages.
- 1Department of Experimental Cardiology University Medical Center Utrecht Utrecht University Utrecht the Netherlands.
- 2CDL Research University Medical Center Utrecht Utrecht the Netherlands.
- 3Regenerative Medicine Centre Utrecht Circulatory Health Research Center University Utrecht University Medical Centre Utrecht Utrecht the Netherlands.
Abstract
Directly post-infarction, myocardial tissue release signals that trigger an acute inflammatory response to clear debris and initiate cardiac repair. However, prolonged inflammation may drive adverse remodelling and heart failure in the long-term. Therefore, understanding the mechanisms of acute and chronic inflammation after myocardial infarction (MI) is crucial for developing targeted treatments. Extracellular vesicles (EVs) mediate intercellular communication by carrying diverse biomolecules, such as proteins, that influence recipient cell behaviour. Cardiomyocyte-derived EVs (CM-EVs) are known to play a role in both cardiac repair and the progression of heart failure following MI. However, little is known about CM-EV subpopulations and how they modulate post-infarction inflammation. Here, we demonstrate that human CM-EV subpopulations can modulate macrophage phenotype. We identified three EV subpopulations, defined by their sedimentation coefficients, with distinct density and proteome profiles released by human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The 2K, 10K and 100K pellets, collected by differential ultracentrifugation of the conditioned medium, are enriched, respectively, in sarcomeric components, metabolic proteins and proteins involved in transport to the plasma membrane. Proteomic analysis revealed that hypoxia changes the proteome of EV-containing pellets, enriching for proteins involved in cellular stress responses, such as chaperones, ribosomal, proteasomal and metabolic proteins. Next, we observed that both normoxic and hypoxic CM-EVs are internalized by macrophages, and the hypoxic 10K pellet induces a pro-inflammatory phenotype in these cells. Future research should elucidate whether CM-EVs contribute to excessive inflammation and adverse cardiac remodelling post-MI in vivo and if this can be modulated by blocking the release of specific CM-EV subpopulations.
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BibTeX
@article{viola2026hypoxic,
title = {{Hypoxic Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Release Large Extracellular Vesicles That Promote a Pro-Inflammatory Phenotype in Macrophages.}},
author = {Margarida Viola and Renee G C Maas and Judy M Lowik and Tom A P Driedonks and Ernest Diez Benavente and Luuk van de Schepop and Cor S Seinen and Christian J B Snijders Blok and Saskia C A de Jager and Pieter Vader and Joost P G Sluijter},
journal = {Journal of extracellular biology},
year = {2026},
month = nov,
doi = {10.1002/jex2.70202},
url = {https://europepmc.org/article/MED/42996290},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - Hypoxic Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Release Large Extracellular Vesicles That Promote a Pro-Inflammatory Phenotype in Macrophages.
AU - Margarida Viola
AU - Renee G C Maas
AU - Judy M Lowik
AU - Tom A P Driedonks
AU - Ernest Diez Benavente
AU - Luuk van de Schepop
AU - Cor S Seinen
AU - Christian J B Snijders Blok
AU - Saskia C A de Jager
AU - Pieter Vader
AU - Joost P G Sluijter
PY - 2026
DA - 2026/11/05
JO - Journal of extracellular biology
DO - 10.1002/jex2.70202
UR - https://europepmc.org/article/MED/42996290
AB - Directly post-infarction, myocardial tissue release signals that trigger an acute inflammatory response to clear debris and initiate cardiac repair. However, prolonged inflammation may drive adverse remodelling and heart failure in the long-term. Therefore, understanding the mechanisms of acute and chronic inflammation after myocardial infarction (MI) is crucial for developing targeted treatments. Extracellular vesicles (EVs) mediate intercellular communication by carrying diverse biomolecules, such as proteins, that influence recipient cell behaviour. Cardiomyocyte-derived EVs (CM-EVs) are known to play a role in both cardiac repair and the progression of heart failure following MI. However, little is known about CM-EV subpopulations and how they modulate post-infarction inflammation. Here, we demonstrate that human CM-EV subpopulations can modulate macrophage phenotype. We identified three EV subpopulations, defined by their sedimentation coefficients, with distinct density and proteome profiles released by human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The 2K, 10K and 100K pellets, collected by differential ultracentrifugation of the conditioned medium, are enriched, respectively, in sarcomeric components, metabolic proteins and proteins involved in transport to the plasma membrane. Proteomic analysis revealed that hypoxia changes the proteome of EV-containing pellets, enriching for proteins involved in cellular stress responses, such as chaperones, ribosomal, proteasomal and metabolic proteins. Next, we observed that both normoxic and hypoxic CM-EVs are internalized by macrophages, and the hypoxic 10K pellet induces a pro-inflammatory phenotype in these cells. Future research should elucidate whether CM-EVs contribute to excessive inflammation and adverse cardiac remodelling post-MI in vivo and if this can be modulated by blocking the release of specific CM-EV subpopulations.
SN - 2768-2811
ER -