Blood glutamate EAAT₂-cell grabbing therapy in cerebral ischemia.

Abstract

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. 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. 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. 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.