Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes

Abstract Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication in the heart under homeostatic and pathological conditions, such as myocardial infarction (MI). However, the basic mechanisms driving cardiomyocyte‐derived EV (CM‐EV) production following stress...

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Main Authors: Margarida Viola, Maarten P. Bebelman, Renee G. C. Maas, Willemijn S. deVoogt, Frederik J. Verweij, Cor S. Seinen, Saskia C. A. deJager, Pieter Vader, Dirk Michiel Pegtel, Joost Petrus Gerardus Sluijter
Format: Article
Language:English
Published: Wiley 2024-11-01
Series:Journal of Extracellular Vesicles
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Online Access:https://doi.org/10.1002/jev2.70000
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author Margarida Viola
Maarten P. Bebelman
Renee G. C. Maas
Willemijn S. deVoogt
Frederik J. Verweij
Cor S. Seinen
Saskia C. A. deJager
Pieter Vader
Dirk Michiel Pegtel
Joost Petrus Gerardus Sluijter
author_facet Margarida Viola
Maarten P. Bebelman
Renee G. C. Maas
Willemijn S. deVoogt
Frederik J. Verweij
Cor S. Seinen
Saskia C. A. deJager
Pieter Vader
Dirk Michiel Pegtel
Joost Petrus Gerardus Sluijter
author_sort Margarida Viola
collection DOAJ
description Abstract Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication in the heart under homeostatic and pathological conditions, such as myocardial infarction (MI). However, the basic mechanisms driving cardiomyocyte‐derived EV (CM‐EV) production following stress are poorly understood. In this study, we generated human induced pluripotent stem cell‐derived cardiomyocytes (hiPSC‐CMs) that express NanoLuc‐tetraspanin reporters. These modified hiPSC‐CMs allow for quantification of tetraspanin‐positive CM‐EV secretion from small numbers of cells without the need for time‐consuming EV isolation techniques. We subjected these cells to a panel of small molecules to study their effect on CM‐EV biogenesis and secretion under basal and stress‐associated conditions. We observed that EV biogenesis is context‐dependent in hiPSC‐CMs. Nutrient starvation decreases CM‐EV secretion while hypoxia increases the production of CM‐EVs in a nSmase2‐dependent manner. Moreover, the inflammatory cytokine TNF‐α increased CM‐EV secretion through a process involving NLRP3 inflammasome activation and mTOR signalling. Here, we detailed for the first time the regulatory mechanisms of EV biogenesis in hiPSC‐CMs upon MI‐associated stressors.
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spelling doaj-art-c9ab97ed038247e4810e4fb8a389601c2024-11-29T11:34:28ZengWileyJournal of Extracellular Vesicles2001-30782024-11-011311n/an/a10.1002/jev2.70000Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytesMargarida Viola0Maarten P. Bebelman1Renee G. C. Maas2Willemijn S. deVoogt3Frederik J. Verweij4Cor S. Seinen5Saskia C. A. deJager6Pieter Vader7Dirk Michiel Pegtel8Joost Petrus Gerardus Sluijter9Department of Experimental Cardiology, University Medical Center UtrechtUtrecht UniversityUtrechtThe NetherlandsDepartment of Pathology, Cancer Center AmsterdamVU University Medical CenterAmsterdamThe NetherlandsDepartment of Experimental Cardiology, University Medical Center UtrechtUtrecht UniversityUtrechtThe NetherlandsCDL ResearchUniversity Medical Center UtrechtUtrechtThe NetherlandsDivision of Cell Biology, Neurobiology and BiophysicsUtrecht UniversityUtrechtThe NetherlandsCDL ResearchUniversity Medical Center UtrechtUtrechtThe NetherlandsDepartment of Experimental Cardiology, University Medical Center UtrechtUtrecht UniversityUtrechtThe NetherlandsDepartment of Experimental Cardiology, University Medical Center UtrechtUtrecht UniversityUtrechtThe NetherlandsDepartment of Pathology, Cancer Center AmsterdamVU University Medical CenterAmsterdamThe NetherlandsDepartment of Experimental Cardiology, University Medical Center UtrechtUtrecht UniversityUtrechtThe NetherlandsAbstract Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication in the heart under homeostatic and pathological conditions, such as myocardial infarction (MI). However, the basic mechanisms driving cardiomyocyte‐derived EV (CM‐EV) production following stress are poorly understood. In this study, we generated human induced pluripotent stem cell‐derived cardiomyocytes (hiPSC‐CMs) that express NanoLuc‐tetraspanin reporters. These modified hiPSC‐CMs allow for quantification of tetraspanin‐positive CM‐EV secretion from small numbers of cells without the need for time‐consuming EV isolation techniques. We subjected these cells to a panel of small molecules to study their effect on CM‐EV biogenesis and secretion under basal and stress‐associated conditions. We observed that EV biogenesis is context‐dependent in hiPSC‐CMs. Nutrient starvation decreases CM‐EV secretion while hypoxia increases the production of CM‐EVs in a nSmase2‐dependent manner. Moreover, the inflammatory cytokine TNF‐α increased CM‐EV secretion through a process involving NLRP3 inflammasome activation and mTOR signalling. Here, we detailed for the first time the regulatory mechanisms of EV biogenesis in hiPSC‐CMs upon MI‐associated stressors.https://doi.org/10.1002/jev2.70000cardiomyocytesEV biogenesisextracellular vesiclesmyocardial infarctionNanoLuc
spellingShingle Margarida Viola
Maarten P. Bebelman
Renee G. C. Maas
Willemijn S. deVoogt
Frederik J. Verweij
Cor S. Seinen
Saskia C. A. deJager
Pieter Vader
Dirk Michiel Pegtel
Joost Petrus Gerardus Sluijter
Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes
Journal of Extracellular Vesicles
cardiomyocytes
EV biogenesis
extracellular vesicles
myocardial infarction
NanoLuc
title Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes
title_full Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes
title_fullStr Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes
title_full_unstemmed Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes
title_short Hypoxia and TNF‐alpha modulate extracellular vesicle release from human induced pluripotent stem cell‐derived cardiomyocytes
title_sort hypoxia and tnf alpha modulate extracellular vesicle release from human induced pluripotent stem cell derived cardiomyocytes
topic cardiomyocytes
EV biogenesis
extracellular vesicles
myocardial infarction
NanoLuc
url https://doi.org/10.1002/jev2.70000
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