Blood glutamate EAAT2-cell grabbing therapy in cerebral ischemiaResearch in context

• Published in: EBioMedicine 2019-01, Vol.39, p.118-131
• Main Authors: María Pérez-Mato, Ramón Iglesias-Rey, Alba Vieites-Prado, Antonio Dopico-López, Bárbara Argibay, Héctor Fernández-Susavila, Andrés da Silva-Candal, Amparo Pérez-Díaz, Clara Correa-Paz, Anne Günther, Paulo Ávila-Gómez, M. Isabel Loza, Arnd Baumann, José Castillo, Tomás Sobrino, Francisco Campos
• Format: Article
• Language: English
• ISSN: 2352-3964; 2352-3964

Background: Excitatory amino acid transporter 2 (EAAT2) plays a pivotal role in glutamate clearance in the adult brain, thereby preventing excitotoxic effects. Considering the high efficacy of EAAT2 for glutamate uptake, we hypothesized that the expression of this transporter in mesenchymal stem cells (MSCs) for systemic administration could yield a cell-based glutamate-grabbing therapy, combining the intrinsic properties of these cells with excitotoxic protection. Methods: To address this hypothesis, EAAT2-encoding cDNA was introduced into MSCs and human embryonic kidney 293 cells (HEK cells) as the control cell line. EAAT2 expression and functionality were evaluated by in vitro assays. Blood glutamate-grabbing activity was tested in healthy and ischemic rat models treated with 3 × 106 and 9 × 106 cells/animal. Findings: The expression of EAAT2 in both cell types conferred the expected glutamate-grabbing activity in in vitro and in vivo studies. The functional improvement observed in ischemic rats treated with EAAT2–HEK at low dose, confirmed that this effect was indeed mediated by the glutamate-grabbing activity associated with EAAT2 functionality. Unexpectedly, both cell doses of non-transfected MSCs induced higher protection than transfected EAAT2–MSCs by another mechanism independent of the glutamate-grabbing capacity. Interpretation: Although the transfection procedure most likely interferes with some of the intrinsic protective mechanisms of mesenchymal cells, the results show that the induced expression of EAAT2 in cells represents a novel alternative to mitigate the excitotoxic effects of glutamate and paves the way to combine this strategy with current cell therapies for cerebral ischemia. Keywords: Cell therapy, Cerebral ischemia, Excitatory amino acid transporter 2, Excitotoxic injury, Excitotoxic protection, Glutamate, Mesenchymal stem cells