AAVs targeting human carbonic anhydrase IV enhance gene delivery to the brain
Gene therapies using natural adeno-associated virus (AAV) serotypes have restricted applications, particularly in the brain, due to their poor targeting and resulting safety concerns. Directed evolution has identified brain-enhanced engineered AAV capsids for various model organisms, but inter-species differences challenge their translation. Here, we engineer AAVs to target human carbonic anhydrase IV (CA-IV), a recently identified blood-brain barrier transcytosis receptor with several favorable properties. We perform in vitro on-column selection of an AAV library to exclude capsids that do not target human CA-IV. We subsequently screen the top 0.01% of variants in vivo in mice expressing human CA-IV in brain endothelial cells. Notably, two top-performing capsids in "humanized" mice are relatively lower ranked in vitro and outperform several top-ranked in vitro candidates. One of these, AAV-hCA4-IV77, achieves 100-fold-greater brain transduction than AAV9, with robust neuronal and astrocytic coverage across brain regions. These results advance our understanding of receptor-targeted capsid design and support the therapeutic potential of human CA-IV-engaging AAVs.
Authors
- Timothy F. Shay (ORCID: https://orcid.org/0000-0001-6591-3271)
- Erin E. Sullivan (ORCID: https://orcid.org/0000-0002-1724-6520)
- Changfan Lin (ORCID: https://orcid.org/0000-0002-8119-9845)
- Bill Kavvathas
- Jonathan D. Hoang (ORCID: https://orcid.org/0000-0002-3762-9596)
- Viviana Gradinaru (ORCID: https://orcid.org/0000-0001-5868-348X)
- Jin Hyung Alex Chung (ORCID: https://orcid.org/0000-0003-2965-9530)
- Seongmin Jang (ORCID: https://orcid.org/0000-0002-9822-6790)
- Tomasz Gawda (ORCID: https://orcid.org/0000-0001-7906-0097)
- Yitong Li (ORCID: https://orcid.org/0000-0003-3402-4484)
- Yujie Fan (ORCID: https://orcid.org/0000-0001-8642-0695)
- Xinhong Chen (ORCID: https://orcid.org/0000-0003-0408-0813)
- Andrew D. Steele
- Irene Tran
- Fiona Ristic
Institutions
- California Institute of Technology (US)
- California State Polytechnic University (US)
Publication Details
- Journal
- Cell Reports
- Published
- 2025-10-31
- DOI
- https://doi.org/10.1016/j.celrep.2025.116419
- Citations
- 5
- Primary Topic
- Virus-based gene therapy research
- Type
- article
- Field-Weighted Citation Impact
- 9.09
Funders
- Friedreich's Ataxia Research Alliance
- National Institutes of Health
- National Heart, Lung, and Blood Institute
- National Institute of Mental Health
- National Institute on Drug Abuse
- National Human Genome Research Institute
- National Cancer Institute
- National Institute of Neurological Disorders and Stroke
- Common Fund