Navarrete, CarmenGarcia-Martin, AdelaCorrea-Saez, AlejandroPrados, Maria E.Fernandez, FranciscoPineda, RafaelMazzone, MassimilianoAlvarez-Benito, MarinaCalzado, Marco AMuñoz, Eduardo2023-05-032023-05-032022-06-30Navarrete C, García-Martín A, Correa-Sáez A, Prados ME, Fernández F, Pineda R, et al. A cannabidiol aminoquinone derivative activates the PP2A/B55α/HIF pathway and shows protective effects in a murine model of traumatic brain injury. J Neuroinflammation. 2022 Jul 9;19(1):177http://hdl.handle.net/10668/20333Traumatic brain injury (TBI) is characterized by a primary mechanical injury and a secondary injury associated with neuroinflammation, blood-brain barrier (BBB) disruption and neurodegeneration. We have developed a novel cannabidiol aminoquinone derivative, VCE-004.8, which is a dual PPARγ/CB2 agonist that also activates the hypoxia inducible factor (HIF) pathway. VCE-004.8 shows potent antifibrotic, anti-inflammatory and neuroprotective activities and it is now in Phase II clinical trials for systemic sclerosis and multiple sclerosis. Herein, we investigated the mechanism of action of VCE-004.8 in the HIF pathway and explored its efficacy in a preclinical model of TBI. Using a phosphoproteomic approach, we investigated the effects of VCE-004.8 on prolyl hydroxylase domain-containing protein 2 (PHD2) posttranslational modifications. The potential role of PP2A/B55α in HIF activation was analyzed using siRNA for B55α. To evaluate the angiogenic response to the treatment with VCE-004.8 we performed a Matrigel plug in vivo assay. Transendothelial electrical resistance (TEER) as well as vascular cell adhesion molecule 1 (VCAM), and zonula occludens 1 (ZO-1) tight junction protein expression were studied in brain microvascular endothelial cells. The efficacy of VCE-004.8 in vivo was evaluated in a controlled cortical impact (CCI) murine model of TBI. Herein we provide evidence that VCE-004.8 inhibits PHD2 Ser125 phosphorylation and activates HIF through a PP2A/B55α pathway. VCE-004.8 induces angiogenesis in vivo increasing the formation of functional vessel (CD31/α-SMA) and prevents in vitro blood-brain barrier (BBB) disruption ameliorating the loss of ZO-1 expression under proinflammatory conditions. In CCI model VCE-004.8 treatment ameliorates early motor deficits after TBI and attenuates cerebral edema preserving BBB integrity. Histopathological analysis revealed that VCE-004.8 treatment induces neovascularization in pericontusional area and prevented immune cell infiltration to the brain parenchyma. In addition, VCE-004.8 attenuates neuroinflammation and reduces neuronal death and apoptosis in the damaged area. This study provides new insight about the mechanism of action of VCE-004.8 regulating the PP2A/B55α/PHD2/HIF pathway. Furthermore, we show the potential efficacy for TBI treatment by preventing BBB disruption, enhancing angiogenesis, and ameliorating neuroinflammation and neurodegeneration after brain injury.enAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/Brain–blood barrierHypoxia-inducible factorNeuroprotectionProlyl-hydroxylasesProtein phosphatase 2ATraumatic brain injuryAnimalsBlood-Brain BarrierBrain Injuries, TraumaticCannabidiolDisease Models, AnimalEndothelial CellsMiceNeovascularization, Pathologicresearch article35810304open access10.1186/s12974-022-02540-91742-2094PMC9270745https://jneuroinflammation.biomedcentral.com/counter/pdf/10.1186/s12974-022-02540-9https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9270745/pdf