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Assessment of AAV9 distribution and transduction in rats after administration through Intrastriatal, Intracisterna magna and Lumbar Intrathecal routes

Abstract

Challenges in obtaining efficient transduction of brain and spinal cord following systemic AAV delivery have led to alternative administration routes being used in clinical trials that directly infuse the virus into the CNS. However, data comparing different direct AAV injections into the brain remain limited making it difficult to choose optimal routes. Here we tested both AAV9-egfp and AAV9-fLuc delivery via intrastriatal (IST), intracisterna magna (ICM) and lumbar intrathecal (LIT) routes in adult rats and assessed vector distribution and transduction in brain, spinal cord and peripheral tissues. We find that IST infusion leads to robust transgene expression in the striatum, thalamus and cortex with lower peripheral tissue transduction and anti-AAV9 capsid titers compared to ICM or LIT. ICM delivery provided strong GFP and luciferase expression across more brain regions than the other routes and similar expression in the spinal cord to LIT injections, which itself largely failed to transduce the rat brain. Our data highlight the strengths and weaknesses of each direct CNS delivery route which will help with future clinical targeting.

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Fig. 1: Overview of injection routes, viral dose, and transgenes expressed for this study.
Fig. 2: AAV9-CAG-fLuc transduction in rat in brain and peripheral tissues.
Fig. 3: Biodistribution of AAV9 vector in rat tissue following intrastriatal (IST), intracisterna magna (ICM) and intrathecal lumbar (LIT) delivery.
Fig. 4: GFP expression in rat brain following intrastriatal (IST), intracisterna magna (ICM) or intrathecal (LIT) administration of AAV9-GFP.
Fig. 5: AAV9-GFP transduces mostly neurons in the rat brain.
Fig. 6: Comparing antibody response to AAV9 capsid in blood and CSF in rats.

Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Funding

This work has been funded by Astrazeneca, UK.

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Contributions

Experiment design: JC, TJ, GMT, MP, CD. Data analysis and interpretation: JC, EAC, DS, SW, GMT, MP. Manuscript preparation: EAC, JC, GMT. Supervision: GMT, MP, DKS, and IC.

Corresponding author

Correspondence to Guy M. L. Meno-Tetang.

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Competing interests

TJ, GMT, CD, DS, IC, and MP are employees of Astrazeneca, UK. JC is an employee of Asklepios Biopharmaceutical, Inc. EAC is an Astrazeneca sponsored postdoctoral fellow & DKS and SW are collaborators working at the University at Buffalo.

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Chandran, J., Chowdhury, E.A., Perkinton, M. et al. Assessment of AAV9 distribution and transduction in rats after administration through Intrastriatal, Intracisterna magna and Lumbar Intrathecal routes. Gene Ther (2022). https://doi.org/10.1038/s41434-022-00346-1

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