Engineering viral vectors for acoustically targeted gene delivery
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1.
California Institute of Technology
Abstract
Targeted gene delivery to the brain is a critical tool for neuroscience research and has significant potential to treat human disease. However, the site-specific delivery of common gene vectors such as adeno-associated viruses (AAVs) is typically performed via invasive injections, which limit its applicable scope of research and clinical applications. Alternatively, focused ultrasound blood-brain-barrier opening (FUS-BBBO), performed noninvasively, enables the site-specific entry of AAVs into the brain from systemic circulation. However, when used in conjunction with natural AAV serotypes, this approach has limited transduction efficiency and results in substantial undesirable transduction of peripheral organs. Here, we use high throughput in vivo selection to engineer new AAV vectors specifically designed for local neuronal transduction at the site of FUS-BBBO. The resulting vectors substantially enhance ultrasound-targeted gene delivery and neuronal tropism while reducing peripheral transduction, providing a more than ten-fold improvement in targeting specificity in two tested mouse strains. In addition to enhancing the only known approach to noninvasively target gene delivery to specific brain regions, these results establish the ability of AAV vectors to be evolved for specific physical delivery mechanisms.
Copyright and License
© The Author(s) 2024. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Acknowledgement
The authors thank Drs. Benjamin Deverman, Nicholas Flytzanis, Nicholas Goeden, and Viviana Gradinaru, and the CLOVER center at Caltech for helpful discussions and the Biological Imaging Facility of the Beckman Institute. This research was supported by the National Institutes of Health (grant UG3MH120102 to M.G.S.), the Jacobs Institute for Molecular Engineering in Medicine, the Sontag Foundation, the Merkin Institute for Translational Research, and 2019 NARSAD Young Investigator Grant from the Brain and Behavior Research Foundation (grant 27737 to J.O.S.). Related work in the Shapiro Lab is supported by the David and Lucille Packard Foundation and the Heritage Medical Research Institute and in Szablowski lab by The G. Harold and Leila Y. Mathers Charitable Foundation. M.G.S. is an Investigator of the Howard Hughes Medical Institute. JEH acknowledges support from Rose Hills foundation and Barry Goldwater Scholarship and from the NSF GRFP. MH acknowledges support from NSF GRFP.
Contributions
H.L., J.O.S., and M.G.S. conceived and planned the research. H.L., M.H., and J.O.S. performed the in vivo experiments with additional input from J.E.H. J.O.S., H.L., and J.E.H. performed the in vitro experiments. J.E.H. and J.O.S. processed the next-generation sequencing data. H.L., M.H., and J.S.T. processed and analyzed histological image data. J.O.S. and M.G.S. wrote the manuscript with input from all other authors. M.G.S. and J.O.S. supervised the research.
Data Availability
The authors declare that all data supporting the results in this study are available within the paper, its Supplementary Information and its Source Data file. Microscopy images and raw sequencing data are available from the corresponding author upon reasonable request owing to their large size and numbers. The NGS data generated through this study have been deposite in the Sequence Read Archive (SRA) database under accession code: PRJNA1112439. Source data are provided with this paper.
Conflict of Interest
J.O.S., M.G.S., J.E.H., and H.L. are inventors on the patent application US20230047753A1. Other authors declare no competing interests.
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Additional details
- National Institutes of Health
- UG3MH120102
- California Institute of Technology
- Jacobs Institute for Molecular Engineering in Medicine
- Sontag Foundation
- California Institute of Technology
- Richard N. Merkin Institute for Translational Medicine
- Brain & Behavior Research Foundation
- 22737
- David and Lucile Packard Foundation
- California Institute of Technology
- Heritage Medical Research Institute
- G. Harold & Leila Y. Mathers Foundation
- Howard Hughes Medical Institute
- Rose Hills Foundation
- National Science Foundation
- NSF Graduate Research Fellowship DGE-2146755
- Caltech groups
- Richard N. Merkin Institute for Translational Research, Jacobs Institute for Molecular Engineering for Medicine, Heritage Medical Research Institute