Endothelial-Derived Extracellular Vesicles Induce Cerebrovascular Dysfunction in Inflammation
David Roig-Carles,
Eduard Willms,
Ruud D. Fontijn,
Sarai Martinez-Pacheco,
Imre Mäger,
Helga E. de Vries,
Mark Hirst,
Basil Sharrack,
David K. Male,
Cheryl A. Hawkes,
Ignacio A. Romero
Affiliations
David Roig-Carles
School of Life, Health and Chemical Sciences, Biomedical Research Network, Open University, Milton Keynes MK7 6AA, UK
Eduard Willms
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC 3086, Australia
Ruud D. Fontijn
Amsterdam UMC, Department of Molecular Cell Biology and Immunology, MS Centre Amsterdam, de Boelelaan 1117 VU University, 1081 HZ Amsterdam, The Netherlands
Sarai Martinez-Pacheco
School of Life, Health and Chemical Sciences, Biomedical Research Network, Open University, Milton Keynes MK7 6AA, UK
Imre Mäger
Department of Paediatrics, University of Oxford, Oxford OX3 9DU, UK
Helga E. de Vries
Amsterdam UMC, Department of Molecular Cell Biology and Immunology, MS Centre Amsterdam, de Boelelaan 1117 VU University, 1081 HZ Amsterdam, The Netherlands
Mark Hirst
School of Life, Health and Chemical Sciences, Biomedical Research Network, Open University, Milton Keynes MK7 6AA, UK
Basil Sharrack
Academic Department of Neuroscience and Sheffield NIHR Translational Neuroscience BRC, Sheffield Teaching Hospitals, NHS Foundation Trust, University of Sheffield, Sheffield S10 2JF, UK
David K. Male
School of Life, Health and Chemical Sciences, Biomedical Research Network, Open University, Milton Keynes MK7 6AA, UK
Cheryl A. Hawkes
Department of Biomedical and Life Sciences, Lancaster University, Lancaster LA1 4YW, UK
Ignacio A. Romero
School of Life, Health and Chemical Sciences, Biomedical Research Network, Open University, Milton Keynes MK7 6AA, UK
Blood–brain barrier (BBB) dysfunction is a key hallmark in the pathology of many neuroinflammatory disorders. Extracellular vesicles (EVs) are lipid membrane-enclosed carriers of molecular cargo that are involved in cell-to-cell communication. Circulating endothelial EVs are increased in the plasma of patients with neurological disorders, and immune cell-derived EVs are known to modulate cerebrovascular functions. However, little is known about whether brain endothelial cell (BEC)-derived EVs themselves contribute to BBB dysfunction. Human cerebral microvascular cells (hCMEC/D3) were treated with TNFα and IFNy, and the EVs were isolated and characterised. The effect of EVs on BBB transendothelial resistance (TEER) and leukocyte adhesion in hCMEC/D3 cells was measured by electric substrate cell-substrate impedance sensing and the flow-based T-cell adhesion assay. EV-induced molecular changes in recipient hCMEC/D3 cells were analysed by RT-qPCR and Western blotting. A stimulation of naïve hCMEC/D3 cells with small EVs (sEVs) reduced the TEER and increased the shear-resistant T-cell adhesion. The levels of microRNA-155, VCAM1 and ICAM1 were increased in sEV-treated hCMEC/D3 cells. Blocking the expression of VCAM1, but not of ICAM1, prevented sEV-mediated T-cell adhesion to brain endothelia. These results suggest that sEVs derived from inflamed BECs promote cerebrovascular dysfunction. These findings may provide new insights into the mechanisms involving neuroinflammatory disorders.
