Blood glutamate EAAT2-cell grabbing therapy in cerebral ischemia

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. This study was partially supported by grants from Instituto de Salud Carlos III (PI13/00292 and PI17/0054), Spanish Research Network on Cerebrovascular Diseases RETICS-INVICTUS (RD12/0014), Fundación Mutua Madrileña; the Ministry of Economy and Competitiveness of Spain (SAF2014-56336-R), Xunta de Galicia (Programa de Desarrollo Precomercial de los resultados de investigación del Sistema Público de Salud de Galicia_ PRIS); and the European Union program FEDER. Furthermore, F. Campos (CP14/00154) and T. Sobrino (CP12/03121 and CPII17/00027) are recipients of a research contract from Miguel Servet Program of Instituto de Salud Carlos III SI