Canonical Wnt Pathway Maintains Blood-Brain Barrier Integrity upon Ischemic Stroke and Its Activation Ameliorates Tissue Plasminogen Activator Therapy
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Stroke induces blood-brain barrier (BBB) breakdown, which promotes complications like oedema and hemorrhagic transformation. Administration of recombinant tissue plasminogen activator (rtPA) within a therapeutic time window of 4.5 h after stroke onset constitutes the only existing treatment. Beyond this time window, rtPA worsens BBB breakdown. Canonical Wnt pathway induces BBB formation and maturation during ontogeny. We hypothesized that the pathway is required to maintain BBB functions after stroke; thus, its activation might improve rtPA therapy. Therefore, we first assessed pathway activity in the brain of mice subjected to transient middle cerebral artery occlusion (MCAo). Next, we evaluated the effect of pathway deactivation early after stroke onset on BBB functions. Finally, we assessed the impact of pathway activation on BBB breakdown associated to delayed administration of rtPA. Our results show that pathway activity is induced predominately in endothelial cells early after ischemic stroke. Early deactivation of the pathway using a potent inhibitor, XAV939, aggravates BBB breakdown and increases hemorrhagic transformation incidence. On the other hand, pathway activation using a potent activator, 6-bromoindirubin-3′-oxime (6-BIO), reduces the incidence of hemorrhagic transformation associated to delayed rtPA administration by attenuating BBB breakdown via promotion of tight junction formation and repressing endothelial basal permeability independently of rtPA proteolytic activity. BBB preservation upon pathway activation limited the deleterious effects of delayed rtPA administration. Our study demonstrates that activation of the canonical Wnt pathway constitutes a clinically relevant strategy to extend the therapeutic time window of rtPA by attenuating BBB breakdown via regulation of BBB-specific mechanisms.
KeywordsIschemic stroke Hemorrhagic transformation Blood-brain barrier Brain endothelial cells Tissue plasminogen activator Canonical Wnt pathway Tight junctions
We would like to thank Mrs. Revathy Guruswamy for performing some of the MCAo surgeries, Dr. Nathalie Vernoux for preparing the brain samples for TEM, and Mrs. Julie-Christine Lévesque at the Bio-Imaging Platform of CRCHU de Québec for technical assistance.
This study was financially supported by grants from the Natural Sciences and Engineering Research Council of Canada (NSERC), Heart and Stroke Foundation of Canada (HSFC), and Fonds de recherche du Québec - Santé (FRQS) (all to AEA). NJL was supported by a scholarship from the NSERC, and a scholarship from the Fondation du CHU de Québec. KP was supported by a scholarship from the Fondation du CHU de Québec. MET is a Canada Research Chair (Tier 2) in Neuroimmune Plasticity in Health and Therapy.
Compliance with Ethical Standards
Animal experiments were performed according to the Canadian Council on Animal Care guidelines, as administered by the Université Laval Animal Welfare Committee.
Conflict of Interest
The authors declare that they have no conflict of interest.
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