Pilot Study Assessing the Impact of Intrathecal Administration of Variants AAV-PHP.B and AAV-PHP.eB on Brain Transduction in Adult Rhesus Macaques.

Marie-Laure Arotcarena,Erwan Bezard,Nathalie Dutheil,Sandra Dovero,Vincent Planche,Nathalie Biendon,Benjamin Dehay

doi:10.3389/fbioe.2021.762209

Abstract

Adeno-associated virus (AAV) vectors are increasingly used as an effective and safe approach to deliver genetic material to the central nervous system (CNS). The AAV9-derived variants, AAV-PHP. B and AAV-PHP.eB, reportedly broadly transduce cells throughout the CNS compared to the original serotype 9, AAV9. As non-human primate data are scarce, we here evaluated the CNS transduction efficiencies after lumbar intrathecal bolus delivery of identical doses of either AAV-PHP. B:CAG-EGFP or AAV-PHP. eB:CAG-EGFP in rhesus macaque monkeys. AAV-PHP.eB achieved a more efficient and widespread CNS transduction compared to AAV-PHP.B. We report a strong neuronal and oligodendroglial tropism for both variants in the putamen and in the hippocampus. This proof-of-concept experiment highlights the potential value of intrathecal infusions of AAV-PHP.eB to distribute genetic material in the CNS with cell-type specificity and introduces a new opportunity to model brain diseases in rhesus macaque monkeys and further develop gene therapies targeting the CNS in humans.

Highlights

The use of non-pathogenic adeno-associated virus (AAV) vectors has emerged as an effective and safe approach for both preclinical modeling and therapeutic methods for neurological disorders (Bourdenx et al, 2014; Deverman et al, 2018; Hudry and Vandenberghe, 2019; Nectow and Nestler, 2020; Fajardo-Serrano et al, 2021)
B (Figure 4A) and AAV-PHP. eB (Figure 4B) conditions, double immunolabeling with the pan-neuronal marker NeuN or the well-established oligodendrocyte marker CNPase showed that EGFP was expressed predominantly in neurons and oligodendrocytes, suggesting a neuronal tropism
To assess whether the neuronal and oligodendroglial tropism observed in the putamen was identical in other brain regions with a different distribution of cell types, we investigated the cellular transgene expression of EGFP in the hippocampus of both the AAVPHP

Summary

Introduction

The use of non-pathogenic adeno-associated virus (AAV) vectors has emerged as an effective and safe approach for both preclinical modeling and therapeutic methods for neurological disorders (Bourdenx et al, 2014; Deverman et al, 2018; Hudry and Vandenberghe, 2019; Nectow and Nestler, 2020; Fajardo-Serrano et al, 2021). Specific serotypes of AAV, such as recombinant adeno-associated virus serotype 9 (AAV9) and pseudotype rhesus-10 (AAVrh.10), have shown unique properties to target the central nervous system (CNS) in comparison to most AAV serotypes (Foust et al, 2009). Since both serotypes share the ability to cross the blood-brain barrier (BBB) in newborns, they may be delivered through various administration routes (Bourdenx et al, 2014; Bedbrook et al, 2018). B did not demonstrate the same capabilities in both marmosets (6 weeks survival) (Matsuzaki et al, 2018) and rhesus macaques (3 weeks survival) (Hordeaux et al, 2018)

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Results

Conclusion