The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress
Abstract The role of GRP94, an endoplasmic reticulum (ER) stress protein with both pro- and anti-inflammatory functions, has not been investigated in macrophages during ER stress, whereas ER stress has been reported in many diseases involving macrophages. In this work, we studied GRP94 in M1/LPS + I...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Publishing Group
2021-01-01
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| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-020-03288-x |
| _version_ | 1818392854288924672 |
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| author | Killian Chaumonnot Sophie Masson Hugo Sikner Alexanne Bouchard Valentin Baverel Pierre-Simon Bellaye Bertrand Collin Carmen Garrido Evelyne Kohli |
| author_facet | Killian Chaumonnot Sophie Masson Hugo Sikner Alexanne Bouchard Valentin Baverel Pierre-Simon Bellaye Bertrand Collin Carmen Garrido Evelyne Kohli |
| author_sort | Killian Chaumonnot |
| collection | DOAJ |
| description | Abstract The role of GRP94, an endoplasmic reticulum (ER) stress protein with both pro- and anti-inflammatory functions, has not been investigated in macrophages during ER stress, whereas ER stress has been reported in many diseases involving macrophages. In this work, we studied GRP94 in M1/LPS + IFNγ and M2/IL-4 primary macrophages derived from human monocytes (isolated from buffy coats), in basal and ER stress conditions induced by thapsigargin (Tg), an inducer of ER calcium depletion and tunicamycin (Tm), an inhibitor of N-glycosylation. We found that GRP94 was expressed on the membrane of M2 but not M1 macrophages. In M2, Tg, but not Tm, while decreased GRP94 content in the membrane, it induced its secretion. This correlated with the induction of a pro-inflammatory profile, which was dependent on the UPR IRE1α arm activation and on a functional GRP94. As we previously reported that GRP94 associated with complement C3 at the extracellular level, we analyzed C3 and confirmed GRP94-C3 interaction in our experimental model. Further, Tg increased this interaction and, in these conditions, C3b and cathepsin L were detected in the extracellular medium where GRP94 co-immunoprecipitated with C3 and C3b. Finally, we showed that the C3b inactivated fragment, iC3b, only present on non-stressed M2, depended on functional GRP94, making both GRP94 and iC3b potential markers of M2 cells. In conclusion, our results show that GRP94 is co-secreted with C3 under ER stress conditions which may facilitate its cleavage by cathepsin L, thus contributing to the pro-inflammatory profile observed in stressed M2 macrophages. |
| first_indexed | 2024-12-14T05:36:02Z |
| format | Article |
| id | doaj.art-c293ab5a1df347eea241df8e1302f7d4 |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-12-14T05:36:02Z |
| publishDate | 2021-01-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-c293ab5a1df347eea241df8e1302f7d42022-12-21T23:15:11ZengNature Publishing GroupCell Death and Disease2041-48892021-01-0112111510.1038/s41419-020-03288-xThe HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stressKillian Chaumonnot0Sophie Masson1Hugo Sikner2Alexanne Bouchard3Valentin Baverel4Pierre-Simon Bellaye5Bertrand Collin6Carmen Garrido7Evelyne Kohli8UMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUFR des Sciences de Santé, Université de BourgogneUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICUMR INSERM/uB/AGROSUP 1231, Team 3 HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTICAbstract The role of GRP94, an endoplasmic reticulum (ER) stress protein with both pro- and anti-inflammatory functions, has not been investigated in macrophages during ER stress, whereas ER stress has been reported in many diseases involving macrophages. In this work, we studied GRP94 in M1/LPS + IFNγ and M2/IL-4 primary macrophages derived from human monocytes (isolated from buffy coats), in basal and ER stress conditions induced by thapsigargin (Tg), an inducer of ER calcium depletion and tunicamycin (Tm), an inhibitor of N-glycosylation. We found that GRP94 was expressed on the membrane of M2 but not M1 macrophages. In M2, Tg, but not Tm, while decreased GRP94 content in the membrane, it induced its secretion. This correlated with the induction of a pro-inflammatory profile, which was dependent on the UPR IRE1α arm activation and on a functional GRP94. As we previously reported that GRP94 associated with complement C3 at the extracellular level, we analyzed C3 and confirmed GRP94-C3 interaction in our experimental model. Further, Tg increased this interaction and, in these conditions, C3b and cathepsin L were detected in the extracellular medium where GRP94 co-immunoprecipitated with C3 and C3b. Finally, we showed that the C3b inactivated fragment, iC3b, only present on non-stressed M2, depended on functional GRP94, making both GRP94 and iC3b potential markers of M2 cells. In conclusion, our results show that GRP94 is co-secreted with C3 under ER stress conditions which may facilitate its cleavage by cathepsin L, thus contributing to the pro-inflammatory profile observed in stressed M2 macrophages.https://doi.org/10.1038/s41419-020-03288-x |
| spellingShingle | Killian Chaumonnot Sophie Masson Hugo Sikner Alexanne Bouchard Valentin Baverel Pierre-Simon Bellaye Bertrand Collin Carmen Garrido Evelyne Kohli The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress Cell Death and Disease |
| title | The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress |
| title_full | The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress |
| title_fullStr | The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress |
| title_full_unstemmed | The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress |
| title_short | The HSP GRP94 interacts with macrophage intracellular complement C3 and impacts M2 profile during ER stress |
| title_sort | hsp grp94 interacts with macrophage intracellular complement c3 and impacts m2 profile during er stress |
| url | https://doi.org/10.1038/s41419-020-03288-x |
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