PMID- 35810304 OWN - NLM STAT- MEDLINE DCOM- 20220712 LR - 20220716 IS - 1742-2094 (Electronic) IS - 1742-2094 (Linking) VI - 19 IP - 1 DP - 2022 Jul 9 TI - A cannabidiol aminoquinone derivative activates the PP2A/B55alpha/HIF pathway and shows protective effects in a murine model of traumatic brain injury. PG - 177 LID - 10.1186/s12974-022-02540-9 [doi] LID - 177 AB - BACKGROUND: Traumatic 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 PPARgamma/CB(2) 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. METHODS: 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/B55alpha in HIF activation was analyzed using siRNA for B55alpha. 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. RESULTS: Herein we provide evidence that VCE-004.8 inhibits PHD2 Ser125 phosphorylation and activates HIF through a PP2A/B55alpha pathway. VCE-004.8 induces angiogenesis in vivo increasing the formation of functional vessel (CD31/alpha-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. CONCLUSIONS: This study provides new insight about the mechanism of action of VCE-004.8 regulating the PP2A/B55alpha/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. CI - (c) 2022. The Author(s). FAU - Navarrete, Carmen AU - Navarrete C AD - Emerald Health Pharmaceuticals, San Diego, USA. FAU - Garcia-Martin, Adela AU - Garcia-Martin A AD - Emerald Health Pharmaceuticals, San Diego, USA. FAU - Correa-Saez, Alejandro AU - Correa-Saez A AD - Maimonides Biomedical Research Institute of Cordoba, University of Cordoba, Avda Menendez Pidal s/n, 14004, Cordoba, Spain. AD - Cellular Biology, Physiology and Immunology Department, University of Cordoba, Cordoba, Spain. AD - Hospital Universitario Reina Sofia, Cordoba, Spain. FAU - Prados, Maria E AU - Prados ME AD - VivaCell Biotechnology Espana, Cordoba, Spain. FAU - Fernandez, Francisco AU - Fernandez F AD - FEA Radiodiagnostico, Seccion de Neurorradiologia Diagnostica. Hospital Universitario Reina Sofia, Cordoba, Spain. FAU - Pineda, Rafael AU - Pineda R AD - Maimonides Biomedical Research Institute of Cordoba, University of Cordoba, Avda Menendez Pidal s/n, 14004, Cordoba, Spain. AD - Cellular Biology, Physiology and Immunology Department, University of Cordoba, Cordoba, Spain. AD - Hospital Universitario Reina Sofia, Cordoba, Spain. FAU - Mazzone, Massimiliano AU - Mazzone M AD - Laboratory of Tumor Inflammation and Angiogenesis, Center for Cancer Biology, VIB-KULeuven, 3000, Louvain, Belgium. FAU - Alvarez-Benito, Marina AU - Alvarez-Benito M AD - Unidad de Radiodiagnostico Y Cancer de Mama, Hospital Universitario Reina Sofia, Cordoba, Spain. FAU - Calzado, Marco A AU - Calzado MA AD - Maimonides Biomedical Research Institute of Cordoba, University of Cordoba, Avda Menendez Pidal s/n, 14004, Cordoba, Spain. AD - Cellular Biology, Physiology and Immunology Department, University of Cordoba, Cordoba, Spain. AD - Hospital Universitario Reina Sofia, Cordoba, Spain. FAU - Munoz, Eduardo AU - Munoz E AD - Emerald Health Pharmaceuticals, San Diego, USA. fi1muble@uco.es. AD - Maimonides Biomedical Research Institute of Cordoba, University of Cordoba, Avda Menendez Pidal s/n, 14004, Cordoba, Spain. fi1muble@uco.es. AD - Cellular Biology, Physiology and Immunology Department, University of Cordoba, Cordoba, Spain. fi1muble@uco.es. AD - Hospital Universitario Reina Sofia, Cordoba, Spain. fi1muble@uco.es. LA - eng GR - PID2020-114753RB-I00/Ministerio de Ciencia e Innovacion/ GR - PID2020-114753RB-I00/Ministerio de Ciencia e Innovacion/ GR - PID2020-114753RB-I00/Ministerio de Ciencia e Innovacion/ GR - P18-RTJ-4163/Junta de A/ PT - Journal Article DEP - 20220709 PL - England TA - J Neuroinflammation JT - Journal of neuroinflammation JID - 101222974 RN - 19GBJ60SN5 (Cannabidiol) SB - IM MH - Animals MH - Blood-Brain Barrier/metabolism MH - *Brain Injuries, Traumatic/complications/drug therapy/metabolism MH - *Cannabidiol MH - Disease Models, Animal MH - Endothelial Cells/metabolism MH - Mice MH - Neovascularization, Pathologic/metabolism PMC - PMC9270745 OTO - NOTNLM OT - Brain-blood barrier OT - Hypoxia-inducible factor OT - Neuroprotection OT - Prolyl-hydroxylases OT - Protein phosphatase 2A OT - Traumatic brain injury COIS- Carmen Navarrete and Adela Garcia-Martin are employees of Emerald Health Pharmaceuticals. Eduardo Munoz is Chief Scientific Officer of Emerald Health Pharmaceuticals. EDAT- 2022/07/10 06:00 MHDA- 2022/07/14 06:00 PMCR- 2022/07/09 CRDT- 2022/07/09 23:25 PHST- 2022/03/07 00:00 [received] PHST- 2022/06/30 00:00 [accepted] PHST- 2022/07/09 23:25 [entrez] PHST- 2022/07/10 06:00 [pubmed] PHST- 2022/07/14 06:00 [medline] PHST- 2022/07/09 00:00 [pmc-release] AID - 10.1186/s12974-022-02540-9 [pii] AID - 2540 [pii] AID - 10.1186/s12974-022-02540-9 [doi] PST - epublish SO - J Neuroinflammation. 2022 Jul 9;19(1):177. doi: 10.1186/s12974-022-02540-9.