Combined gene therapy with vascular endothelial growth factor plus apelin in a chronic cerebral hypoperfusion model in rats

Abstract

OBJECTIVEThe aim of this study was to evaluate whether combined gene therapy with vascular endothelial growth factor (VEGF) plus apelin during indirect vasoreconstructive surgery enhances brain angiogenesis in a chronic cerebral hypoperfusion model in rats.METHODSA chronic cerebral hypoperfusion model induced by the permanent ligation of bilateral common carotid arteries (CCAs; a procedure herein referred to as “CCA occlusion” [CCAO]) in rats was employed in this study. Seven days after the CCAO procedure, the authors performed encephalo-myo-synangiosis (EMS) and injected plasmid(s) into each rat's temporal muscle. Rats were divided into 4 groups based on which plasmid was received (i.e., LacZ group, VEGF group, apelin group, and VEGF+apelin group). Protein levels in the cortex and attached muscle were assessed with enzyme-linked immunosorbent assay (ELISA) on Day 7 after EMS, while immunofluorescent analysis of cortical vessels was performed on Day 14 after EMS.RESULTSThe total number of blood vessels in the cortex on Day 14 after EMS was significantly larger in the VEGF group and the VEGF+apelin group than in the LacZ group (p < 0.05, respectively). Larger vessels appeared in the VEGF+apelin group than in the other groups (p < 0.05, respectively). Apelin protein on Day 7 after EMS was not detected in the cortex for any of the groups. In the attached muscle, apelin protein was detected only in the apelin group and the VEGF+apelin group. Immunofluorescent analysis revealed that apelin and its receptor, APJ, were expressed on endothelial cells (ECs) 7 days after the CCAO.CONCLUSIONSCombined gene therapy (VEGF plus apelin) during EMS in a chronic cerebral hypoperfusion model can enhance angiogenesis in rats. This treatment has the potential to be a feasible option in a clinical setting for patients with moyamoya disease.

High-Mobility Group Box-1-Induced Angiogenesis After Indirect Bypass Surgery in a Chronic Cerebral Hypoperfusion Model

“…Previous experimental reports showed that administration of the plasmid human VEGF with EMS significantly increased the number of vessels compared with EMS surgery alone (Kusaka et al 2005; Katsumata et al 2010). Moreover, in the combination of gene therapy with VEGF plus apelin, the number of vessels was significantly increased compared with EMS alone, and mature vessels were significantly developed compared with EMS alone, or compared with administration of plasmid human VEGF with EMS (Hiramatsu et al 2017). These data suggest that administration of other angiogenic factors with EMS surgery may also enhance angiogenesis, leading to an improvement in cerebral circulation.…”

“…We previously examined the effect of indirect revascularization surgery with vascular endothelial growth factor (VEGF) gene administration into the temporal muscle in a chronic cerebral hypoperfusion state in a rat model (Kusaka et al 2005; Katsumata et al 2010). Moreover, we investigated the effect of combined gene therapy with VEGF plus apelin during indirect revascularization surgery (Hiramatsu et al 2017). EMS for the hypoperfusion state after bilateral common carotid artery (CCA) ligation simulated indirect bypass surgery for patients with MMD.…”

A refined model of chronic cerebral hypoperfusion resulting in cognitive impairment and a low mortality rate in rats

“…14,34 Reportedly, the most prevalent treatment for moyamoya disease, indirect bypass (e.g., encephalomyosynangiosis), could be improved through combination with gene or stem cell therapy. 8,11 As our mCCAO model can be easily combined with indirect bypass or cell therapies, it can facilitate studying the pathophysiology of vascular dementia and moyamoya disease.…”

Impaired functional recovery of endothelial colony-forming cells from moyamoya disease in a chronic cerebral hypoperfusion rat model

“…The previous MMD studies using the CCH model in rats evaluated the effect of gene therapy under the ischemic status induced by the model, but they did not evaluate MMD ECFC function itself. 12 In this study, it is meaningful that the function of MMD ECFCs was evaluated in comparison with the in vivo animal model.…”