RESCUING BLOOD–BRAIN BARRIER GLUCOSE TRANSPORT IN GLUT1-DS USING ENGINEERED EPCS UNDER DYNAMIC FLOW CONDITIONS: A STEP TOWARD CELL-BASED THERAPY

GLUT1 Deficiency Syndrome (GLUT1-DS) is a neurological disorder caused by mutations in the SLC2A1 gene, which impair glucose transport across the blood–brain barrier (BBB), leading to chronic cerebral energy deficiency, developmental delay, epilepsy, movement disorders, and progressive cognitive impairment. While gene replacement is therapeutic, systemic viral delivery carries risks of ectopic targeting. To minimize these risks, we propose a cell-based therapy approach, in which autologous endothelial progenitor cells (EPCs) are genetically corrected ex vivo via lentiviral vectors to overexpress functional GLUT1. This study focuses on validating this "living drug" strategy using a biomimetic, dynamic in vitro BBB model. To replicate physiological shear stress and barrier maturation, we utilize the IVTech Technologies fluidic platform. Currently, we have successfully optimized the experimental protocol using human brain microvascular endothelial cells (hBMEC) to establish a stable baseline of barrier integrity, monitored through the expression of key junctional markers and permeability assays. The core of our investigation follows a sequential methodology: first, we will integrate patient-derived EPCs to reconstruct a pathological BBB model; subsequently, lentivirally-corrected EPCs will be introduced into the circuit to evaluate their capacity to integrate into the endothelial monolayer and restore glucose flux. Preliminary results indicate that the IVTech Technologies dynamic environment significantly enhances barrier tightness compared to static cultures. We conclude that using corrected EPCs for targeted GLUT1 delivery represents a promising strategy for GLUT1-DS, offering a proof-of-concept for a personalized medicine approach that bypasses the limitations of traditional gene therapy.