title: AAV9-mediated central nervous system-targeted gene delivery via cisterna magna route in mice.
abstract: Current barriers to the use of adeno-associated virus serotype 9 (AAV9) in clinical trials for treating neurological disorders are its high expression in many off-target tissues such as liver and heart, and lack of cell specificity within the central nervous system (CNS) when using ubiquitous promoters such as human cytomegalovirus (CMV) or chicken-β-actin hybrid (CAG). To enhance targeting the transgene expression in CNS cells, self-complementary (sc) AAV9 vectors, scAAV9-GFP vectors carrying neuronal Hb9 and synapsin 1, and nonspecific CMV and CAG promoters were constructed. We demonstrate that synapsin 1 and Hb9 promoters exclusively targeted neurons in vitro, although their strengths were up to 10-fold lower than that of CMV. In vivo analyses of mouse tissue after scAAV9-GFP vector delivery via the cisterna magna revealed a significant advantage of synapsin 1 promoter over both Hb9 variants in targeting neurons throughout the brain, since Hb9 promoters were driving gene expression mainly within the motor-related areas of the brain stem. In summary, this study demonstrates that cisterna magna administration is a safe alternative to intracranial or intracerebroventricular vector delivery route using scAAV9, and introduces a novel utility of the Hb9 promoter for the targeted gene expression for both in vivo and in vitro applications.
authors.affiliation: Sheffield Institute for Translational Neuroscience, Department of Neuroscience, University of Sheffield , Sheffield, UK.
authors.forename: Vera
authors.initials: V
authors.lastname: Lukashchuk
authors.affiliation: Sheffield Institute for Translational Neuroscience, Department of Neuroscience, University of Sheffield , Sheffield, UK.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Lewis
authors.affiliation: Sheffield Institute for Translational Neuroscience, Department of Neuroscience, University of Sheffield , Sheffield, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Coldicott
authors.affiliation: Sheffield Institute for Translational Neuroscience, Department of Neuroscience, University of Sheffield , Sheffield, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Grierson
authors.affiliation: Sheffield Institute for Translational Neuroscience, Department of Neuroscience, University of Sheffield, Sheffield, UK; Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Mimoun
authors.initials: M
authors.lastname: Azzouz