Cargando…
A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation
AIMS: Endothelial hyperpermeability exacerbates multiple organ damage during inflammation or infection. The endothelial glycocalyx, a protective matrix covering the luminal surface of endothelial cells (ECs), undergoes enzymatic shedding during inflammation, contributing to barrier hyperpermeability...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6198742/ https://www.ncbi.nlm.nih.gov/pubmed/29939250 http://dx.doi.org/10.1093/cvr/cvy167 |
_version_ | 1783365017259737088 |
---|---|
author | Yang, Xiaoyuan Meegan, Jamie E Jannaway, Melanie Coleman, Danielle C Yuan, Sarah Y |
author_facet | Yang, Xiaoyuan Meegan, Jamie E Jannaway, Melanie Coleman, Danielle C Yuan, Sarah Y |
author_sort | Yang, Xiaoyuan |
collection | PubMed |
description | AIMS: Endothelial hyperpermeability exacerbates multiple organ damage during inflammation or infection. The endothelial glycocalyx, a protective matrix covering the luminal surface of endothelial cells (ECs), undergoes enzymatic shedding during inflammation, contributing to barrier hyperpermeability. A disintegrin and metalloproteinase 15 (ADAM15) is a sheddase capable of cleaving the ectodomains of membrane-bound molecules. Herein, we tested whether and how ADAM15 is involved in glycocalyx shedding and vascular leakage during sepsis. METHODS AND RESULTS: Dextran-150kD exclusion assay revealed lipopolysaccharide (LPS) significantly reduced glycocalyx thickness in mouse cremaster microvessels. Consistently, shedding products of glycocalyx constituents, including CD44 ectodomain, were detected with an increased plasma level after cecal ligation and puncture (CLP)-induced sepsis. The direct effects of CD44 ectodomain on endothelial barrier function were evaluated, which revealed CD44 ectodomain dose-dependently reduced transendothelial electrical resistance (TER) and caused cell–cell adherens junction disorganization. Furthermore, we examined the role of ADAM15 in CD44 cleavage and glycocalyx shedding. An in vitro cleavage assay coupled with liquid chromatography-tandem mass spectrometry confirmed ADAM15 cleaved CD44 at His(235)-Thr(236) bond. In ECs with ADAM15 knockdown, LPS-induced CD44 cleavage and TER reduction were greatly attenuated, whereas, ADAM15 overexpression exacerbated CD44 cleavage and TER response to LPS. Consistently, ADAM15 knockout in mice attenuated CLP-induced increase in plasma CD44. Intravital and electron microscopic images revealed ADAM15 deficiency prevented LPS-induced glycocalyx injury in cremaster and pulmonary microvasculatures. Functionally, ADAM15(−/−) mice with better-preserved glycocalyx exhibited resistance to LPS-induced vascular leakage, as evidenced by reduced albumin extravasation in pulmonary and mesenteric vessels. Importantly, in intact, functionally vital human lungs, perfusion of LPS induced a significant up-regulation of ADAM15, accompanied by elevated CD44 in the effluent and increased vascular permeability to albumin. CONCLUSION: Together, our data support the critical role of ADAM15 in mediating vascular barrier dysfunction during inflammation. Its mechanisms of action involve CD44 shedding and endothelial glycocalyx injury. |
format | Online Article Text |
id | pubmed-6198742 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-61987422018-10-26 A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation Yang, Xiaoyuan Meegan, Jamie E Jannaway, Melanie Coleman, Danielle C Yuan, Sarah Y Cardiovasc Res Original Articles AIMS: Endothelial hyperpermeability exacerbates multiple organ damage during inflammation or infection. The endothelial glycocalyx, a protective matrix covering the luminal surface of endothelial cells (ECs), undergoes enzymatic shedding during inflammation, contributing to barrier hyperpermeability. A disintegrin and metalloproteinase 15 (ADAM15) is a sheddase capable of cleaving the ectodomains of membrane-bound molecules. Herein, we tested whether and how ADAM15 is involved in glycocalyx shedding and vascular leakage during sepsis. METHODS AND RESULTS: Dextran-150kD exclusion assay revealed lipopolysaccharide (LPS) significantly reduced glycocalyx thickness in mouse cremaster microvessels. Consistently, shedding products of glycocalyx constituents, including CD44 ectodomain, were detected with an increased plasma level after cecal ligation and puncture (CLP)-induced sepsis. The direct effects of CD44 ectodomain on endothelial barrier function were evaluated, which revealed CD44 ectodomain dose-dependently reduced transendothelial electrical resistance (TER) and caused cell–cell adherens junction disorganization. Furthermore, we examined the role of ADAM15 in CD44 cleavage and glycocalyx shedding. An in vitro cleavage assay coupled with liquid chromatography-tandem mass spectrometry confirmed ADAM15 cleaved CD44 at His(235)-Thr(236) bond. In ECs with ADAM15 knockdown, LPS-induced CD44 cleavage and TER reduction were greatly attenuated, whereas, ADAM15 overexpression exacerbated CD44 cleavage and TER response to LPS. Consistently, ADAM15 knockout in mice attenuated CLP-induced increase in plasma CD44. Intravital and electron microscopic images revealed ADAM15 deficiency prevented LPS-induced glycocalyx injury in cremaster and pulmonary microvasculatures. Functionally, ADAM15(−/−) mice with better-preserved glycocalyx exhibited resistance to LPS-induced vascular leakage, as evidenced by reduced albumin extravasation in pulmonary and mesenteric vessels. Importantly, in intact, functionally vital human lungs, perfusion of LPS induced a significant up-regulation of ADAM15, accompanied by elevated CD44 in the effluent and increased vascular permeability to albumin. CONCLUSION: Together, our data support the critical role of ADAM15 in mediating vascular barrier dysfunction during inflammation. Its mechanisms of action involve CD44 shedding and endothelial glycocalyx injury. Oxford University Press 2018-11-01 2018-06-23 /pmc/articles/PMC6198742/ /pubmed/29939250 http://dx.doi.org/10.1093/cvr/cvy167 Text en © The Author(s) 2018. Published by Oxford University Press on behalf of the European Society of Cardiology http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Original Articles Yang, Xiaoyuan Meegan, Jamie E Jannaway, Melanie Coleman, Danielle C Yuan, Sarah Y A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
title | A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
title_full | A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
title_fullStr | A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
title_full_unstemmed | A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
title_short | A disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
title_sort | disintegrin and metalloproteinase 15-mediated glycocalyx shedding contributes to vascular leakage during inflammation |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6198742/ https://www.ncbi.nlm.nih.gov/pubmed/29939250 http://dx.doi.org/10.1093/cvr/cvy167 |
work_keys_str_mv | AT yangxiaoyuan adisintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT meeganjamiee adisintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT jannawaymelanie adisintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT colemandaniellec adisintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT yuansarahy adisintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT yangxiaoyuan disintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT meeganjamiee disintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT jannawaymelanie disintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT colemandaniellec disintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation AT yuansarahy disintegrinandmetalloproteinase15mediatedglycocalyxsheddingcontributestovascularleakageduringinflammation |