Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis
Abstract Apoptosis of endothelial cells prompts the release of apoptotic exosome-like vesicles (ApoExos), subtype extracellular vesicles secreted by apoptotic cells after caspase-3 activation. ApoExos are different from both apoptotic bodies and classical exosomes in their protein and nucleic acid c...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2023-07-01
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| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-023-05991-x |
| _version_ | 1827894847004999680 |
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| author | Alexandre Brodeur Francis Migneault Maude Lanoie Déborah Beillevaire Julie Turgeon Annie Karakeussian-Rimbaud Nicolas Thibodeau Éric Boilard Mélanie Dieudé Marie-Josée Hébert |
| author_facet | Alexandre Brodeur Francis Migneault Maude Lanoie Déborah Beillevaire Julie Turgeon Annie Karakeussian-Rimbaud Nicolas Thibodeau Éric Boilard Mélanie Dieudé Marie-Josée Hébert |
| author_sort | Alexandre Brodeur |
| collection | DOAJ |
| description | Abstract Apoptosis of endothelial cells prompts the release of apoptotic exosome-like vesicles (ApoExos), subtype extracellular vesicles secreted by apoptotic cells after caspase-3 activation. ApoExos are different from both apoptotic bodies and classical exosomes in their protein and nucleic acid contents and functions. In contrast to classical apoptotic bodies, ApoExos induce immunogenic responses that can be maladaptive when not tightly regulated. In the present study, we elucidated the mechanisms by which ApoExos are internalized by endothelial cells, which leads to shared specific and functional mRNAs of importance to endothelial function. Using flow cytometry and confocal microscopy, we revealed that ApoExos were actively internalized by endothelial cells. SiRNA-induced inhibition of classical endocytosis pathways with pharmacological inhibitors showed that ApoExos were internalized via phosphatidylserine-dependent macropinocytosis independently of classical endocytosis pathways. An electron microscopy analysis revealed that ApoExos increased the macropinocytosis rate in endothelial cells, setting in motion a positive feedback loop that increased the amount of internalized ApoExos. Deep sequencing of total RNA revealed that ApoExos possessed a unique protein-coding RNA profile, with PCSK5 being the most abundant mRNA. Internalization of ApoExos by cells led to the transfer of this RNA content from the ApoExos to cells. Specifically, PCSK5 mRNA was transferred to cells that had taken up ApoExos, and these cells subsequently expressed PCSK5. Collectively, our findings suggest that macropinocytosis is an effective entry pathway for the delivery of RNAs carried by ApoExos and that these RNAs are functionally expressed by the endothelial cells that internalize them. As ApoExos express a specific mRNA signature, these results suggest new avenues to understand how ApoExos produced at sites of vascular injury impact vascular function. |
| first_indexed | 2024-03-12T22:13:20Z |
| format | Article |
| id | doaj.art-d82037779e7540eb82097fb4963629ad |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-03-12T22:13:20Z |
| publishDate | 2023-07-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-d82037779e7540eb82097fb4963629ad2023-07-23T11:27:55ZengNature Publishing GroupCell Death and Disease2041-48892023-07-0114711510.1038/s41419-023-05991-xApoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosisAlexandre Brodeur0Francis Migneault1Maude Lanoie2Déborah Beillevaire3Julie Turgeon4Annie Karakeussian-Rimbaud5Nicolas Thibodeau6Éric Boilard7Mélanie Dieudé8Marie-Josée Hébert9Centre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCanadian Donation and Transplantation Research Program (CDTRP)Centre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalCentre de Recherche, Centre Hospitalier de l’Université de Montréal (CRCHUM) and Université de MontréalAbstract Apoptosis of endothelial cells prompts the release of apoptotic exosome-like vesicles (ApoExos), subtype extracellular vesicles secreted by apoptotic cells after caspase-3 activation. ApoExos are different from both apoptotic bodies and classical exosomes in their protein and nucleic acid contents and functions. In contrast to classical apoptotic bodies, ApoExos induce immunogenic responses that can be maladaptive when not tightly regulated. In the present study, we elucidated the mechanisms by which ApoExos are internalized by endothelial cells, which leads to shared specific and functional mRNAs of importance to endothelial function. Using flow cytometry and confocal microscopy, we revealed that ApoExos were actively internalized by endothelial cells. SiRNA-induced inhibition of classical endocytosis pathways with pharmacological inhibitors showed that ApoExos were internalized via phosphatidylserine-dependent macropinocytosis independently of classical endocytosis pathways. An electron microscopy analysis revealed that ApoExos increased the macropinocytosis rate in endothelial cells, setting in motion a positive feedback loop that increased the amount of internalized ApoExos. Deep sequencing of total RNA revealed that ApoExos possessed a unique protein-coding RNA profile, with PCSK5 being the most abundant mRNA. Internalization of ApoExos by cells led to the transfer of this RNA content from the ApoExos to cells. Specifically, PCSK5 mRNA was transferred to cells that had taken up ApoExos, and these cells subsequently expressed PCSK5. Collectively, our findings suggest that macropinocytosis is an effective entry pathway for the delivery of RNAs carried by ApoExos and that these RNAs are functionally expressed by the endothelial cells that internalize them. As ApoExos express a specific mRNA signature, these results suggest new avenues to understand how ApoExos produced at sites of vascular injury impact vascular function.https://doi.org/10.1038/s41419-023-05991-x |
| spellingShingle | Alexandre Brodeur Francis Migneault Maude Lanoie Déborah Beillevaire Julie Turgeon Annie Karakeussian-Rimbaud Nicolas Thibodeau Éric Boilard Mélanie Dieudé Marie-Josée Hébert Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis Cell Death and Disease |
| title | Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis |
| title_full | Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis |
| title_fullStr | Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis |
| title_full_unstemmed | Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis |
| title_short | Apoptotic exosome-like vesicles transfer specific and functional mRNAs to endothelial cells by phosphatidylserine-dependent macropinocytosis |
| title_sort | apoptotic exosome like vesicles transfer specific and functional mrnas to endothelial cells by phosphatidylserine dependent macropinocytosis |
| url | https://doi.org/10.1038/s41419-023-05991-x |
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