Dual targeting of astrocytic and endothelial GLUT1 enables functional rescue in GLUT1 deficiency syndrome

Abstract

Glucose transporter 1 deficiency syndrome (GLUT1-DS) is a severe neurometabolic disorder caused by impaired brain glucose transport. Because GLUT1 has been classically viewed as predominantly localized to blood-brain barrier endothelial cells, gene therapy strategies have largely focused on endothelial GLUT1 restoration. However, whether this endothelial-centered model fully explains disease pathogenesis and therapeutic rescue has remained unclear. Using enhanced detection in mouse brain and human postmortem tissue, we show that GLUT1 is broadly expressed in astrocytes as well as endothelial cells, extending beyond perivascular endfeet into astrocytic somata and processes. Conditional Glut1 haploinsufficiency in either astrocytes or endothelial cells reproduced GLUT1-DS-like reductions in cerebrospinal fluid glucose availability and brain parenchymal glucose responses, together with cognitive and motor impairments. To determine the therapeutic consequence of this revised cellular model, we used complementary endothelial- and astrocyte-directed AAV delivery to restore GLUT1 in systemic Glut1 haploinsufficient mice. Single-compartment restoration produced partial benefit, whereas dual-compartment restoration more robustly improved cerebrospinal fluid glucose availability and cognitive function. These findings indicate that endothelial GLUT1 restoration alone is insufficient for robust rescue and redefine GLUT1-DS as a multicellular disorder of the neurovascular unit.