Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death
Therapy-induced presentation of cell surface calreticulin (CRT) is a pro-phagocytic immunogen beneficial for invoking anti-tumor immunity. Here, we characterized the roles of ERp57 and α-integrins as CRT-interacting proteins that coordinately regulate CRT translocation from the ER to the surface dur...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-05-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fonc.2019.00411/full |
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| author | Chi-Chao Liu Pascal Leclair Foujan Pedari Heidi Vieira Mahdis Monajemi Laura M. Sly Gregor S. Reid Gregor S. Reid Chinten James Lim Chinten James Lim |
| author_facet | Chi-Chao Liu Pascal Leclair Foujan Pedari Heidi Vieira Mahdis Monajemi Laura M. Sly Gregor S. Reid Gregor S. Reid Chinten James Lim Chinten James Lim |
| author_sort | Chi-Chao Liu |
| collection | DOAJ |
| description | Therapy-induced presentation of cell surface calreticulin (CRT) is a pro-phagocytic immunogen beneficial for invoking anti-tumor immunity. Here, we characterized the roles of ERp57 and α-integrins as CRT-interacting proteins that coordinately regulate CRT translocation from the ER to the surface during immunogenic cell death. Using T-lymphoblasts as a genetic cell model, we found that drug-induced surface CRT is dependent on ERp57, while drug-induced surface ERp57 is independent of CRT. Differential subcellular immunostaining assays revealed that ERp57−/− cells have minimal cytosolic CRT, indicating that ERp57 is indispensable for extra-ER accumulation of CRT. Stimulation of integrin activity, with either cell adhesion or molecular agonists, resulted in decreased drug-induced surface CRT and ERp57 levels. Similarly, surface CRT and ERp57 was reduced in cells expressing GFFKR, a conserved α-integrin cytosolic motif that binds CRT. Drug-induced surface ERp57 levels were consistently higher in CRT−/− cells, suggesting integrin inhibition of surface ERp57 is an indirect consequence of α-integrin binding to CRT within the CRT-ERp57 complex. Furthermore, β1−/− cells with reduced expression of multiple α-integrins, exhibit enhanced levels of drug-induced surface CRT and ERp57. Our findings highlight the coordinate involvement of plasma membrane integrins as inhibitors, and ERp57 originating from the ER as promoters, of CRT translocation from the ER to the cell surface. |
| first_indexed | 2024-12-16T15:35:37Z |
| format | Article |
| id | doaj.art-aec8cc951b8744268b6cb660dd7df2f2 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-12-16T15:35:37Z |
| publishDate | 2019-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-aec8cc951b8744268b6cb660dd7df2f22022-12-21T22:26:11ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2019-05-01910.3389/fonc.2019.00411448971Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell DeathChi-Chao Liu0Pascal Leclair1Foujan Pedari2Heidi Vieira3Mahdis Monajemi4Laura M. Sly5Gregor S. Reid6Gregor S. Reid7Chinten James Lim8Chinten James Lim9Department of Pediatrics, University of British Columbia, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaMichael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, Vancouver, BC, CanadaDepartment of Pediatrics, University of British Columbia, Vancouver, BC, CanadaMichael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, Vancouver, BC, CanadaTherapy-induced presentation of cell surface calreticulin (CRT) is a pro-phagocytic immunogen beneficial for invoking anti-tumor immunity. Here, we characterized the roles of ERp57 and α-integrins as CRT-interacting proteins that coordinately regulate CRT translocation from the ER to the surface during immunogenic cell death. Using T-lymphoblasts as a genetic cell model, we found that drug-induced surface CRT is dependent on ERp57, while drug-induced surface ERp57 is independent of CRT. Differential subcellular immunostaining assays revealed that ERp57−/− cells have minimal cytosolic CRT, indicating that ERp57 is indispensable for extra-ER accumulation of CRT. Stimulation of integrin activity, with either cell adhesion or molecular agonists, resulted in decreased drug-induced surface CRT and ERp57 levels. Similarly, surface CRT and ERp57 was reduced in cells expressing GFFKR, a conserved α-integrin cytosolic motif that binds CRT. Drug-induced surface ERp57 levels were consistently higher in CRT−/− cells, suggesting integrin inhibition of surface ERp57 is an indirect consequence of α-integrin binding to CRT within the CRT-ERp57 complex. Furthermore, β1−/− cells with reduced expression of multiple α-integrins, exhibit enhanced levels of drug-induced surface CRT and ERp57. Our findings highlight the coordinate involvement of plasma membrane integrins as inhibitors, and ERp57 originating from the ER as promoters, of CRT translocation from the ER to the cell surface.https://www.frontiersin.org/article/10.3389/fonc.2019.00411/fullintegrinscalreticulinERp57immunogenic cell deathleukemiacell adhesion |
| spellingShingle | Chi-Chao Liu Pascal Leclair Foujan Pedari Heidi Vieira Mahdis Monajemi Laura M. Sly Gregor S. Reid Gregor S. Reid Chinten James Lim Chinten James Lim Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death Frontiers in Oncology integrins calreticulin ERp57 immunogenic cell death leukemia cell adhesion |
| title | Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death |
| title_full | Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death |
| title_fullStr | Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death |
| title_full_unstemmed | Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death |
| title_short | Integrins and ERp57 Coordinate to Regulate Cell Surface Calreticulin in Immunogenic Cell Death |
| title_sort | integrins and erp57 coordinate to regulate cell surface calreticulin in immunogenic cell death |
| topic | integrins calreticulin ERp57 immunogenic cell death leukemia cell adhesion |
| url | https://www.frontiersin.org/article/10.3389/fonc.2019.00411/full |
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