1521-22542042018AprJ Gene MedGene therapy for Glut1-deficient mouse using an adeno-associated virus vector with the human intrinsic GLUT1 promoter.e3013e301310.1002/jgm.3013We generated an adeno-associated virus (AAV) vector in which the human SLC2A1 gene, encoding glucose transporter type 1 (GLUT1), was expressed under the human endogenous GLUT1 promoter (AAV-GLUT1). We examined whether AAV-GLUT1 administration could lead to functional improvement in GLUT1-deficient mice.We extrapolated human endogenous GLUT1 promoter sequences from rat minimal Glut1 promoter sequences. We generated a tyrosine-mutant AAV9/3 vector in which human SLC2A1-myc-DDK was expressed under the human GLUT1 promoter (AAV-GLUT1). AAV-GLUT1 was administered to GLUT1-deficient mice (GLUT1^+/- mice) via intracerebroventricular injection (1.85 × 10¹⁰ vg/mouse or 6.5 × 10¹⁰ vg/mouse). We analyzed exogenous GLUT1 mRNA and protein expression in the brain and other major organs. We also examined improvements of cerebral microvasculature, motor function using rota-rod and footprint tests, as well as blood and cerebrospinal fluid (CSF) glucose levels. Additionally, we confirmed exogenous GLUT1 protein distribution in the brain and other organs after intracardiac injection (7.8 × 10¹¹ vg/mouse).Exogenous GLUT1 protein was strongly expressed in the cerebral cortex, hippocampus and thalamus. It was mainly expressed in endothelial cells, and partially expressed in neural cells and oligodendrocytes. Motor function and CSF glucose levels were significantly improved following intracerebroventricular injection. Exogenous GLUT1 expression was not detected in other organs after intracerebroventricular injection of AAV-GLUT1, whereas it was detected in the liver and muscle tissue after intracardiac injection.Exogenous GLUT1 expression after AAV-GLUT1 injection approximated that of physiological human GLUT1 expression. Local central nervous system administration of AAV-GLUT1 improved CSF glucose levels and motor function of GLUT1-deficient mice and minimized off-target effects.Copyright © 2018 John Wiley & Sons, Ltd.NakamuraSachieSDepartment of Pediatrics, Jichi Medical University, Tochigi, Japan.MuramatsuShin-IchiSIDivision of Neurology, Jichi Medical University, Tochigi, Japan.Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Japan.TakinoNaomiNDivision of Neurology, Jichi Medical University, Tochigi, Japan.ItoMikaMDivision of Neurology, Jichi Medical University, Tochigi, Japan.JimboEriko FEFDepartment of Pediatrics, Jichi Medical University, Tochigi, Japan.ShimazakiKunikoKDepartment of Neurosurgery, Jichi Medical University, Tochigi, Japan.OnakaTatsushiTDivision of Brain and Neurophysiology, Department of Physiology, Jichi Medical University, Tochigi, Japan.OhtsukiSumioSDepartment of Pharmaceutical Microbiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.TerasakiTetsuyaTDivision of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.YamagataTakanoriTDepartment of Pediatrics, Jichi Medical University, Tochigi, Japan.OsakaHitoshiHDepartment of Pediatrics, Jichi Medical University, Tochigi, Japan.engJournal ArticleResearch Support, Non-U.S. Gov't20180406