Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses
Abstract Endothelial cells (ECs) are constantly submitted in vivo to hemodynamical forces derived from the blood circulation, including shear stress (SS). ECs are able to detect SS and consequently adapt their phenotype, thus affecting many endothelial functions. If a plethora of shear stress-regula...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-06-01
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| Series: | Fluids and Barriers of the CNS |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12987-022-00344-w |
| _version_ | 1811241933073285120 |
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| author | Nina Choublier Meryam Taghi Marie-Claude Menet Morgane Le Gall Johanna Bruce Philippe Chafey François Guillonneau Amélie Moreau Claire Denizot Yannick Parmentier Samir Nakib Didier Borderie Haniaa Bouzinba-Segard Pierre-Olivier Couraud Sandrine Bourdoulous Xavier Declèves |
| author_facet | Nina Choublier Meryam Taghi Marie-Claude Menet Morgane Le Gall Johanna Bruce Philippe Chafey François Guillonneau Amélie Moreau Claire Denizot Yannick Parmentier Samir Nakib Didier Borderie Haniaa Bouzinba-Segard Pierre-Olivier Couraud Sandrine Bourdoulous Xavier Declèves |
| author_sort | Nina Choublier |
| collection | DOAJ |
| description | Abstract Endothelial cells (ECs) are constantly submitted in vivo to hemodynamical forces derived from the blood circulation, including shear stress (SS). ECs are able to detect SS and consequently adapt their phenotype, thus affecting many endothelial functions. If a plethora of shear stress-regulated molecular networks have been described in peripheral ECs, less is known about the molecular responses of microvascular brain ECs which constitute the blood–brain barrier (BBB). In this work, we investigated the response of human cerebral microvascular ECs to laminar physiological shear stress using the well characterized hCMEC/D3 cell line. Interestingly, we showed that hCMEC/D3 cells responded to shear stress by aligning perpendicularly to the flow direction, contrary to peripheral endothelial cells which aligned in the flow direction. Whole proteomic profiles were compared between hCMEC/D3 cells cultured either in static condition or under 5 or 10 dyn.cm−2 SS for 3 days. 3592 proteins were identified and expression levels were significantly affected for 3% of them upon both SS conditions. Pathway analyses were performed which revealed that most proteins overexpressed by SS refer to the antioxidant defense, probably mediated by activation of the NRF2 transcriptional factor. Regarding down-regulated proteins, most of them participate to the pro-inflammatory response, cell motility and proliferation. These findings confirm the induction of EC quiescence by laminar physiological SS and reveal a strong protective effect of SS on hCMEC/D3 cells, suggesting a similar effect on the BBB. Our results also showed that SS did not significantly increase expression levels nor did it affect the localization of junctional proteins and did not afect either the functional activity of several ABC transporters (P-glycoprotein and MRPs). This work provides new insights on the response of microvascular brain ECs to SS and on the importance of SS for optimizing in vitro BBB models. |
| first_indexed | 2024-04-12T13:43:33Z |
| format | Article |
| id | doaj.art-19fc31ee25ba4de8bf040d26425522eb |
| institution | Directory Open Access Journal |
| issn | 2045-8118 |
| language | English |
| last_indexed | 2024-04-12T13:43:33Z |
| publishDate | 2022-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Fluids and Barriers of the CNS |
| spelling | doaj.art-19fc31ee25ba4de8bf040d26425522eb2022-12-22T03:30:45ZengBMCFluids and Barriers of the CNS2045-81182022-06-0119112410.1186/s12987-022-00344-wExposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responsesNina Choublier0Meryam Taghi1Marie-Claude Menet2Morgane Le Gall3Johanna Bruce4Philippe Chafey5François Guillonneau6Amélie Moreau7Claire Denizot8Yannick Parmentier9Samir Nakib10Didier Borderie11Haniaa Bouzinba-Segard12Pierre-Olivier Couraud13Sandrine Bourdoulous14Xavier Declèves15INSERM, Optimisation Thérapeutique en Neuropsychopharmacologie, Université Paris CitéINSERM, Optimisation Thérapeutique en Neuropsychopharmacologie, Université Paris CitéInstitut de Chimie Physique, CNRS, Université Paris-Saclay3P5 Proteom’IC Facility, Institut Cochin, INSERM, CNRS, Université de Paris3P5 Proteom’IC Facility, Institut Cochin, INSERM, CNRS, Université de Paris3P5 Proteom’IC Facility, Institut Cochin, INSERM, CNRS, Université de Paris3P5 Proteom’IC Facility, Institut Cochin, INSERM, CNRS, Université de ParisTechnologie ServierTechnologie ServierTechnologie ServierService de Biochimie, Hôpital Cochin, Assistance Publique-Hôpitaux de ParisService de Biochimie, Hôpital Cochin, Assistance Publique-Hôpitaux de ParisCNRS, INSERM, Institut Cochin, Inserm, CNRS, Université Paris CitéCNRS, INSERM, Institut Cochin, Inserm, CNRS, Université Paris CitéCNRS, INSERM, Institut Cochin, Inserm, CNRS, Université Paris CitéINSERM, Optimisation Thérapeutique en Neuropsychopharmacologie, Université Paris CitéAbstract Endothelial cells (ECs) are constantly submitted in vivo to hemodynamical forces derived from the blood circulation, including shear stress (SS). ECs are able to detect SS and consequently adapt their phenotype, thus affecting many endothelial functions. If a plethora of shear stress-regulated molecular networks have been described in peripheral ECs, less is known about the molecular responses of microvascular brain ECs which constitute the blood–brain barrier (BBB). In this work, we investigated the response of human cerebral microvascular ECs to laminar physiological shear stress using the well characterized hCMEC/D3 cell line. Interestingly, we showed that hCMEC/D3 cells responded to shear stress by aligning perpendicularly to the flow direction, contrary to peripheral endothelial cells which aligned in the flow direction. Whole proteomic profiles were compared between hCMEC/D3 cells cultured either in static condition or under 5 or 10 dyn.cm−2 SS for 3 days. 3592 proteins were identified and expression levels were significantly affected for 3% of them upon both SS conditions. Pathway analyses were performed which revealed that most proteins overexpressed by SS refer to the antioxidant defense, probably mediated by activation of the NRF2 transcriptional factor. Regarding down-regulated proteins, most of them participate to the pro-inflammatory response, cell motility and proliferation. These findings confirm the induction of EC quiescence by laminar physiological SS and reveal a strong protective effect of SS on hCMEC/D3 cells, suggesting a similar effect on the BBB. Our results also showed that SS did not significantly increase expression levels nor did it affect the localization of junctional proteins and did not afect either the functional activity of several ABC transporters (P-glycoprotein and MRPs). This work provides new insights on the response of microvascular brain ECs to SS and on the importance of SS for optimizing in vitro BBB models.https://doi.org/10.1186/s12987-022-00344-wHuman endothelial cellsProteomicsShear stressBlood–brain barrierBrain microvessels |
| spellingShingle | Nina Choublier Meryam Taghi Marie-Claude Menet Morgane Le Gall Johanna Bruce Philippe Chafey François Guillonneau Amélie Moreau Claire Denizot Yannick Parmentier Samir Nakib Didier Borderie Haniaa Bouzinba-Segard Pierre-Olivier Couraud Sandrine Bourdoulous Xavier Declèves Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses Fluids and Barriers of the CNS Human endothelial cells Proteomics Shear stress Blood–brain barrier Brain microvessels |
| title | Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses |
| title_full | Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses |
| title_fullStr | Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses |
| title_full_unstemmed | Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses |
| title_short | Exposure of human cerebral microvascular endothelial cells hCMEC/D3 to laminar shear stress induces vascular protective responses |
| title_sort | exposure of human cerebral microvascular endothelial cells hcmec d3 to laminar shear stress induces vascular protective responses |
| topic | Human endothelial cells Proteomics Shear stress Blood–brain barrier Brain microvessels |
| url | https://doi.org/10.1186/s12987-022-00344-w |
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