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Acoustically targeted noninvasive gene therapy in large brain volumes

Abstract

Focused Ultrasound Blood-Brain Barrier Opening (FUS-BBBO) can deliver adeno-associated viral vectors (AAVs) to treat genetic disorders of the brain. However, such disorders often affect large brain regions. Moreover, the applicability of FUS-BBBO in the treatment of brain-wide genetic disorders has not yet been evaluated. Herein, we evaluated the transduction efficiency and safety of opening up to 105 sites simultaneously. Increasing the number of targeted sites increased gene delivery efficiency at each site. We achieved transduction of up to 60% of brain cells with comparable efficiency in the majority of the brain regions. Furthermore, gene delivery with FUS-BBBO was safe even when all 105 sites were targeted simultaneously without negative effects on animal weight or neuronal loss. To evaluate the application of multi-site FUS-BBBO for gene therapy, we used it for gene editing using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system and found effective gene editing, but also a loss of neurons at the targeted sites. Overall, this study provides a brain-wide map of transduction efficiency, shows the synergistic effect of multi-site targeting on transduction efficiency, and is the first example of large brain volume gene editing after noninvasive gene delivery with FUS-BBBO.

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Fig. 1: Achieving effective & safe FUS-BBBO over large brain areas for extensive AAV-mediated gene delivery.
Fig. 2: Gene delivery efficiency increases with the volume of BBB opened.
Fig. 3: Widespread gene delivery with FUS-BBBO.
Fig. 4: Long-term safety of FUS-BBBO in multiple brain sites.
Fig. 5: Gene editing after noninvasive gene delivery to multiple brain sites.

Data availability

Numerical data is available in the attached source data file. Images used in the generation of this data are available upon reasonable request.

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Acknowledgements

The authors thank Dr. Mingshan Xue (Baylor College of Medicine) for helpful discussions, and Gang Bao’s laboratory (Rice University) for providing the usage of the ultracentrifuge. This research was supported by the Dunn Foundation John S. Dunn Foundation award for collaborative research and by The Welch Foundation grant (C-2048-20200401). Related work was funded by Harold Y and G. Leila Mathers Foundation.

Funding

The work was funded by John S. Dunn Foundation award for collaborative research and The Welch Foundation. Related work was supported by and Harold Y and G. Leila Mathers Foundation.

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JOS and SN designed the experiments, SN and SL performed the in vivo studies, JOS and SN wrote the manuscript, SN, VAS and JY performed histological processing, imaging, and counting, HCD and SN performed preliminary in vivo experiments, JOS supervised the study.

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Correspondence to Jerzy O. Szablowski.

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Nouraein, S., Lee, S., Saenz, V.A. et al. Acoustically targeted noninvasive gene therapy in large brain volumes. Gene Ther (2023). https://doi.org/10.1038/s41434-023-00421-1

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