The lack of technology for direct global-scale targeting of the adult mouse nervous system has hindered research on brain processing and dysfunctions. Currently, gene transfer is normally achieved by intraparenchymal viral injections, but these injections target a restricted brain area. Herein, we demonstrated that intravenous delivery of adeno-associated virus (AAV)-PHP.B viral particles permeated and diffused throughout the neural parenchyma, targeting both the central and the peripheral nervous system in a global pattern. We then established multiple procedures of viral transduction to control gene expression or inactivate gene function exclusively in the adult nervous system and assessed the underlying behavioral effects. Building on these results, we established an effective gene therapy strategy to counteract the widespread accumulation of α-synuclein deposits throughout the forebrain in a mouse model of synucleinopathy. Transduction of A53T-SCNA transgenic mice with AAV-PHP.B-GBA1 restored physiological levels of the enzyme, reduced α-synuclein pathology, and produced significant behavioral recovery. Finally, we provided evidence that AAV-PHP.B brain penetration does not lead to evident dysfunctions in blood-brain barrier integrity or permeability. Altogether, the AAV-PHP.B viral platform enables non-invasive, widespread, and long-lasting global neural expression of therapeutic genes, such as GBA1, providing an invaluable approach to treat neurodegenerative diseases with diffuse brain pathology such as synucleinopathies.
La mancanza di tecnologia per la manipolazione diretta su scala globale del sistema nervoso del topo adulto ha ostacolato la ricerca sui processi e disfunzioni cerebrali. Attualmente, il trasferimento genico è normalmente ottenuto mediante iniezioni virali intraparenchimali, ma queste iniezioni mirano a un'area del cervello ristretta. Qui, abbiamo dimostrato che la somministrazione endovenosa di un vettore virale adeno-associato (AAV) -PHP.B ha permeato e diffuso in tutto il parenchima neurale, trasducendo sia il sistema nervoso centrale e periferico in un modello globale. Abbiamo quindi stabilito molteplici procedure di trasduzione virale per controllare l'espressione genica o disattivare la funzione genica esclusivamente nel sistema nervoso adulto e valutato gli effetti comportamentali. Sulla base di questi risultati, abbiamo stabilito un'efficace strategia di terapia genica per contrastare l'accumulo diffuso di depositi di α-sinucleina in tutto il cervello in un modello murino di sinucleinopatia. Trasduzione di topi transgenici A53T-SCNA con AAV-PHP.B-GBA1 ha ripristinato i livelli fisiologici di Glucocerebrosidasi, ha ridotto l’accumulo di α-sinucleina e ha prodotto un significativo recupero comportamentale. Infine, abbiamo fornito prove che la penetrazione cerebrale di AAV-PHP.B non comporta evidenti disfunzioni nell'integrità o permeabilità della barriera ematoencefalica. Complessivamente, la strategia basata sul vettore virale AAV-PHP.B consente l'espressione neurale globale non invasiva, diffusa e duratura di geni terapeutici, come GBA1, fornendo un approccio inestimabile per il trattamento di malattie neurodegenerative con patologia cerebrale diffusa come le sinucleinopatie.
Gene therapy approaches for neurological disorders using newly generated AAV strains
MORABITO, GIUSEPPE
2018
Abstract
The lack of technology for direct global-scale targeting of the adult mouse nervous system has hindered research on brain processing and dysfunctions. Currently, gene transfer is normally achieved by intraparenchymal viral injections, but these injections target a restricted brain area. Herein, we demonstrated that intravenous delivery of adeno-associated virus (AAV)-PHP.B viral particles permeated and diffused throughout the neural parenchyma, targeting both the central and the peripheral nervous system in a global pattern. We then established multiple procedures of viral transduction to control gene expression or inactivate gene function exclusively in the adult nervous system and assessed the underlying behavioral effects. Building on these results, we established an effective gene therapy strategy to counteract the widespread accumulation of α-synuclein deposits throughout the forebrain in a mouse model of synucleinopathy. Transduction of A53T-SCNA transgenic mice with AAV-PHP.B-GBA1 restored physiological levels of the enzyme, reduced α-synuclein pathology, and produced significant behavioral recovery. Finally, we provided evidence that AAV-PHP.B brain penetration does not lead to evident dysfunctions in blood-brain barrier integrity or permeability. Altogether, the AAV-PHP.B viral platform enables non-invasive, widespread, and long-lasting global neural expression of therapeutic genes, such as GBA1, providing an invaluable approach to treat neurodegenerative diseases with diffuse brain pathology such as synucleinopathies.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.14242/106998
URN:NBN:IT:UNIMIB-106998