AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset
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Abstract
Genetic intervention is increasingly being explored as a therapeutic option for debilitating disorders of the central nervous system. The safety and efficacy of gene therapies rely upon expressing a transgene in affected cells while minimizing off-target expression. Here we show organ-specific targeting of adeno-associated virus (AAV) capsids after intravenous delivery, which we achieved by employing a Cre-transgenic-based screening platform and sequential engineering of AAV-PHP.eB between the surface-exposed AA452 and AA460 of VP3. From this selection, we identified capsid variants that were enriched in the brain and targeted away from the liver in C57BL/6J mice. This tropism extends to marmoset (Callithrix jacchus), enabling robust, non-invasive gene delivery to the marmoset brain after intravenous administration. Notably, the capsids identified result in distinct transgene expression profiles within the brain, with one exhibiting high specificity to neurons. The ability to cross the blood-brain barrier with neuronal specificity in rodents and non-human primates enables new avenues for basic research and therapeutic possibilities unattainable with naturally occurring serotypes.
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Bioengineering of novel AAV variants for BBB-crossing and CNS targeting has achieved some successes, yielding AAV.CPP.16, AAV.CAP-B10, and AAV.cc47 with variable enhancements over AAV9 from mice to NHPs.
“The bioengineering of novel AAV variants in terms of BBB-crossing and CNS targeting has achieved some successes in recent years, yielding AAV.CPP.16 [57], AAV.CAP-B10 [68] and AAV.cc47 [111] with variable enhancements over AAV9 from mice to NHPs.”
From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [68]
Later, they optimized the CREATE platform by introducing a peptide library into the surface-exposed AA455 loop of PhP.eB capsid and obtained AAV.CAP-B10 with enhanced CNS tropism and simultaneously de-targeted from liver expression in both mice and marmosets.
“Later, they optimized the CREATE platform by introducing peptide library into the surface-exposed AA455 loop of PhP.eB capsid and obtained a variant AAV.CAP-B10 with enhanced CNS tropism and simultaneously de-targeted from liver expression in both mice and marmosets [68].”
From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [68]
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- Authors
- Goertsen, D., Flytzanis, N. C., Goeden, N., Chuapoco, M. R., Cummins, A., Chen, Y., Fan, Y., Zhang, Q., Sharma, J., Duan, Y., Wang, L., Feng, G., Chen, Y., Ip, N. Y., Pickel, J., & Gradinaru, V.
- Affiliations
- California Institute of Technology (US)
- National Institutes of Health (US)
- Chinese Academy of Sciences (CN)
- Broad Institute (US)
- Harvard University (US)
- Hong Kong University of Science and Technology (HK)
- Corresponding author
- Gradinaru, Viviana
- ORCID
- Venue
- Nature Neuroscience
- Published
- 2021-12-09
- Volume
- 25
- Issue
- 1
- Pages
- 106-115
- ISSN
- 1097-6256 (ISSN-L)
- 1546-1726
- OpenAlex
- W4200543263
- Cited by
- 420
- References
- 13
- Citations by year
- 2026: 70 · 2025: 115 · 2024: 90 · 2023: 82 · 2022: 55 · 2021: 6 · 2020: 2
- FWCI
- 32.94
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- Nature Neuroscience (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- CaltechAUTHORS (California Institute of Technology) (submittedVersion)
- Rare & Special e-Zone (The Hong Kong University of Science and Technology) (submittedVersion)
- Funders
- Gordon and Betty Moore Foundation
- California Institute of Technology
- Vallee Foundation
- Heritage Medical Research Institute
- University Grants Committee (AoE/M-604/16)
- Shenzhen Knowledge Innovation Program (JCYJ20170413165053031)
- National Institutes of Health (DP1OD025535, 1uf1mh128336-01, U24OD026638)
- Advanced Research Projects Agency
- Beckman Institute, California Institute of Technology
- National Institute of Mental Health (RF1MH117069)
- National Institute of Neurological Disorders and Stroke (DP2NS087949)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
- Keywords
- AAV-PHP.eB, brain targeting, liver targeting, intravenous gene delivery, blood-brain barrier penetration, marmoset, central nervous system gene therapy, off-target expression
- MeSH terms
- Dependovirus, Animals, Brain, Callithrix, Capsid, Genetic Vectors, Liver, Mice, Inbred C57BL, Transduction, Genetic, Transgenes, Mice, Administration, Intravenous
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