Circulating small extracellular vesicles mediate vascular hyperpermeability in diabetes

Springer Science and Business Media LLC - 2024
Dakota Gustafson1,2, Peter V. DiStefano1, Xue Fan Wang3, Ruilin Wu1,2, Siavash Ghaffari4, Crizza Ching1,5, Kumaragurubaran Rathnakumar1, Faisal Alibhai1, Michal Syonov4, Jessica Fitzpatrick6, Emilie Boudreau1, Cori Lau1,2, Natalie Galant1, Mansoor Husain1,2, Ren-Ke Li1, Warren L. Lee4,7,2, Rulan S. Parekh6, Philippe P. Monnier3,8,9, Jason E. Fish1,10,2,5
1Toronto General Hospital Research Institute, University Health Network, Toronto, Canada
2Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada
3Division of Fundamental Neurobiology, Toronto Western Research Institute, University Health Network, Toronto, Canada
4Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Toronto, Canada
5Institute of Medical Science, University of Toronto, Toronto, Canada
6Department of Medicine and Pediatrics, Women’s College Hospital, Hospital for Sick Children and University of Toronto, Toronto, Canada
7Department of Biochemistry, University of Toronto, Toronto, Canada
8Department of Physiology, Faculty of Medicine, University of Toronto, Toronto, Canada
9Donald K. Johnson Eye Institute, Krembil Research Institute, University Health Network, Toronto, Canada
10Peter Munk Cardiac Centre, University Health Network, Toronto, Canada

Tóm tắt

A hallmark chronic complication of type 2 diabetes mellitus is vascular hyperpermeability, which encompasses dysfunction of the cerebrovascular endothelium and the subsequent development of associated cognitive impairment. The present study tested the hypothesis that during type 2 diabetes circulating small extracellular vesicles (sEVs) exhibit phenotypic changes that facilitate pathogenic disruption of the vascular barrier. sEVs isolated from the plasma of a mouse model of type 2 diabetes and from diabetic human individuals were characterised for their ability to disrupt the endothelial cell (EC) barrier. The contents of sEVs and their effect on recipient ECs were assessed by proteomics and identified pathways were functionally interrogated with small molecule inhibitors. Using intravital imaging, we found that diabetic mice (Leprdb/db) displayed hyperpermeability of the cerebrovasculature. Enhanced vascular leakiness was recapitulated following i.v. injection of sEVs from diabetic mice into non-diabetic recipient mice. Characterisation of circulating sEV populations from the plasma of diabetic mice and humans demonstrated increased quantity and size of sEVs compared with those isolated from non-diabetic counterparts. Functional experiments revealed that sEVs from diabetic mice or humans induced the rapid and sustained disruption of the EC barrier through enhanced paracellular and transcellular leak but did not induce inflammation. Subsequent sEV proteome and recipient EC phospho-proteome analysis suggested that extracellular vesicles (sEVs) from diabetic mice and humans modulate the MAPK/MAPK kinase (MEK) and Rho-associated protein kinase (ROCK) pathways, cell–cell junctions and actin dynamics. This was confirmed experimentally. Treatment of sEVs with proteinase K or pre-treatment of recipient cells with MEK or ROCK inhibitors reduced the hyperpermeability-inducing effects of circulating sEVs in the diabetic state. Diabetes is associated with marked increases in the concentration and size of circulating sEVs. The modulation of sEV-associated proteins under diabetic conditions can induce vascular leak through activation of the MEK/ROCK pathway. These data identify a new paradigm by which diabetes can induce hyperpermeability and dysfunction of the cerebrovasculature and may implicate sEVs in the pathogenesis of cognitive decline during type 2 diabetes.

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