Year: 2025
Type: article
Abstract: 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.
Source: Cell Reports
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $5,620
Funders: Friedreich's Ataxia Research Alliance, National Institutes of Health, Beckman Institute, California Institute of Technology, 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, Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Common Fund, Merkin Institute for Translational Research, California Institute of Technology
Awards: Broadening access with an Armamentarium Vector Core Powered by Inclusive Research Experiences, Circuit-Specific Delivery of Large Cargo Across the Nervous Systems of Adult Mammals and Embryos via Novel Engineered Systemic Vectors, G5228164454, Engineered AAV Identification, Validation, and Dissemination Pipeline for Brain Cell Type-Specific Manipulation Across Species, G6129623716
OpenAlex ID: W4415729906