Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium
Abstract The intracellular scaffold KRIT1/CCM1 is an established regulator of vascular barrier function. Loss of KRIT1 leads to decreased microvessel barrier function and to the development of the vascular disorder Cerebral Cavernous Malformation (CCM). However, how loss of KRIT1 causes the subseque...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
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Nature Portfolio
2017-08-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-08373-4 |
| _version_ | 1818845706051387392 |
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| author | Luca Goitre Peter V. DiStefano Andrea Moglia Nicholas Nobiletti Eva Baldini Lorenza Trabalzini Julie Keubel Eliana Trapani Vladimir V. Shuvaev Vladimir R. Muzykantov Ingrid H. Sarelius Saverio Francesco Retta Angela J. Glading |
| author_facet | Luca Goitre Peter V. DiStefano Andrea Moglia Nicholas Nobiletti Eva Baldini Lorenza Trabalzini Julie Keubel Eliana Trapani Vladimir V. Shuvaev Vladimir R. Muzykantov Ingrid H. Sarelius Saverio Francesco Retta Angela J. Glading |
| author_sort | Luca Goitre |
| collection | DOAJ |
| description | Abstract The intracellular scaffold KRIT1/CCM1 is an established regulator of vascular barrier function. Loss of KRIT1 leads to decreased microvessel barrier function and to the development of the vascular disorder Cerebral Cavernous Malformation (CCM). However, how loss of KRIT1 causes the subsequent deficit in barrier function remains undefined. Previous studies have shown that loss of KRIT1 increases the production of reactive oxygen species (ROS) and exacerbates vascular permeability triggered by several inflammatory stimuli, but not TNF−α. We now show that endothelial ROS production directly contributes to the loss of barrier function in KRIT1 deficient animals and cells, as targeted antioxidant enzymes reversed the increase in permeability in KRIT1 heterozygous mice as shown by intravital microscopy. Rescue of the redox state restored responsiveness to TNF-α in KRIT1 deficient arterioles, but not venules. In vitro, KRIT1 depletion increased endothelial ROS production via NADPH oxidase signaling, up-regulated Nox4 expression, and promoted NF-κB dependent promoter activity. Recombinant yeast avenanthramide I, an antioxidant and inhibitor of NF-κB signaling, rescued barrier function in KRIT1 deficient cells. However, KRIT1 depletion blunted ROS production in response to TNF-α. Together, our data indicate that ROS signaling is critical for the loss of barrier function following genetic deletion of KRIT1. |
| first_indexed | 2024-12-19T05:33:55Z |
| format | Article |
| id | doaj.art-6fc3adc7ae70457492848b492ff196e1 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-19T05:33:55Z |
| publishDate | 2017-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-6fc3adc7ae70457492848b492ff196e12022-12-21T20:34:10ZengNature PortfolioScientific Reports2045-23222017-08-017111210.1038/s41598-017-08373-4Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endotheliumLuca Goitre0Peter V. DiStefano1Andrea Moglia2Nicholas Nobiletti3Eva Baldini4Lorenza Trabalzini5Julie Keubel6Eliana Trapani7Vladimir V. Shuvaev8Vladimir R. Muzykantov9Ingrid H. Sarelius10Saverio Francesco Retta11Angela J. Glading12Department of Clinical and Biological Sciences, University of TorinoDepartment of Pharmacology and Physiology, University of RochesterDepartment of Agriculture, Forest and Food Sciences, Plant Genetics and Breeding, University of TorinoDepartment of Pharmacology and Physiology, University of RochesterDepartment of Pharmacology and Physiology, University of RochesterDepartment of Biotechnology, Chemistry and Pharmacy, University of SienaDepartment of Pharmacology and Physiology, University of RochesterDepartment of Clinical and Biological Sciences, University of TorinoDepartment of Pharmacology, University of PennsylvaniaDepartment of Pharmacology, University of PennsylvaniaDepartment of Pharmacology and Physiology, University of RochesterDepartment of Clinical and Biological Sciences, University of TorinoDepartment of Pharmacology and Physiology, University of RochesterAbstract The intracellular scaffold KRIT1/CCM1 is an established regulator of vascular barrier function. Loss of KRIT1 leads to decreased microvessel barrier function and to the development of the vascular disorder Cerebral Cavernous Malformation (CCM). However, how loss of KRIT1 causes the subsequent deficit in barrier function remains undefined. Previous studies have shown that loss of KRIT1 increases the production of reactive oxygen species (ROS) and exacerbates vascular permeability triggered by several inflammatory stimuli, but not TNF−α. We now show that endothelial ROS production directly contributes to the loss of barrier function in KRIT1 deficient animals and cells, as targeted antioxidant enzymes reversed the increase in permeability in KRIT1 heterozygous mice as shown by intravital microscopy. Rescue of the redox state restored responsiveness to TNF-α in KRIT1 deficient arterioles, but not venules. In vitro, KRIT1 depletion increased endothelial ROS production via NADPH oxidase signaling, up-regulated Nox4 expression, and promoted NF-κB dependent promoter activity. Recombinant yeast avenanthramide I, an antioxidant and inhibitor of NF-κB signaling, rescued barrier function in KRIT1 deficient cells. However, KRIT1 depletion blunted ROS production in response to TNF-α. Together, our data indicate that ROS signaling is critical for the loss of barrier function following genetic deletion of KRIT1.https://doi.org/10.1038/s41598-017-08373-4 |
| spellingShingle | Luca Goitre Peter V. DiStefano Andrea Moglia Nicholas Nobiletti Eva Baldini Lorenza Trabalzini Julie Keubel Eliana Trapani Vladimir V. Shuvaev Vladimir R. Muzykantov Ingrid H. Sarelius Saverio Francesco Retta Angela J. Glading Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium Scientific Reports |
| title | Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium |
| title_full | Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium |
| title_fullStr | Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium |
| title_full_unstemmed | Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium |
| title_short | Up-regulation of NADPH oxidase-mediated redox signaling contributes to the loss of barrier function in KRIT1 deficient endothelium |
| title_sort | up regulation of nadph oxidase mediated redox signaling contributes to the loss of barrier function in krit1 deficient endothelium |
| url | https://doi.org/10.1038/s41598-017-08373-4 |
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