An alternate receptor for adeno-associated viruses

Bijay P Dhungel; Hua Xu; Rajini Nagarajah; Joseph Vitale; Alex C H Wong; Divya Gokal; Yue Feng; Mehdi Sharifi Tabar; Cynthia Metierre; Chirag Parsania; Xiaohui Song; Guopeng Wang; Xiao-Dong Su; Charles G Bailey; John E J Rasko

doi:10.1016/j.cell.2025.06.026

An alternate receptor for adeno-associated viruses

Cell. 2025 Sep 4;188(18):4924-4935.e23. doi: 10.1016/j.cell.2025.06.026. Epub 2025 Jul 14.

Authors

Affiliations

¹ School of Medical Sciences, Faculty of Medicine & Health, The University of Sydney, Camperdown, NSW 2006, Australia; Cancer & Gene Regulation Laboratory Centenary Institute, The University of Sydney, Camperdown, NSW 2050, Australia; Centre for Rare Diseases & Gene Therapy Centenary Institute, The University of Sydney, Camperdown, NSW 2050, Australia.
² School of Life Sciences, Peking University, Beijing, China; Biomedical Pioneering Innovation Center (BIOPIC), Peking University, Beijing, China; State Key Laboratory of Protein and Plant Gene Research, Peking University, Beijing, China.
³ Cancer & Gene Regulation Laboratory Centenary Institute, The University of Sydney, Camperdown, NSW 2050, Australia; Centre for Rare Diseases & Gene Therapy Centenary Institute, The University of Sydney, Camperdown, NSW 2050, Australia.
⁴ School of Life Sciences, Peking University, Beijing, China.
⁵ School of Life Sciences, Peking University, Beijing, China; Biomedical Pioneering Innovation Center (BIOPIC), Peking University, Beijing, China; State Key Laboratory of Protein and Plant Gene Research, Peking University, Beijing, China. Electronic address: xdsu@pku.edu.cn.
⁶ School of Medical Sciences, Faculty of Medicine & Health, The University of Sydney, Camperdown, NSW 2006, Australia; Cancer & Gene Regulation Laboratory Centenary Institute, The University of Sydney, Camperdown, NSW 2050, Australia; Centre for Rare Diseases & Gene Therapy Centenary Institute, The University of Sydney, Camperdown, NSW 2050, Australia. Electronic address: charles.bailey@sydney.edu.au.
⁷ School of Medical Sciences, Faculty of Medicine & Health, The University of Sydney, Camperdown, NSW 2006, Australia; Cell and Molecular Therapies, Royal Prince Alfred Hospital, Camperdown, NSW 2050, Australia. Electronic address: jejrasko@gmail.com.

PMID: 40664211
DOI: 10.1016/j.cell.2025.06.026

Abstract

Systemic gene therapy using adeno-associated virus (AAV) vectors is approved for the treatment of several genetic disorders, but challenges and toxicities associated with high vector doses remain. We report an alternate receptor for AAV (AAVR2, carboxypeptidase D [CPD]), which is distinct from the multi-serotype AAV receptor (AAVR). AAVR2 enables the transduction of clade E AAVs, including AAV8, and determines an exclusive AAVR-independent transduction pathway for AAV11 and AAV12. We characterized direct binding between the AAV8 capsid and AAVR2 by cryo-electron microscopy (cryo-EM) and identified contact residues. We observed that AAV8 directly binds to the carboxypeptidase-like domain 1 of AAVR2 via its variable region VIII and demonstrated that AAV capsids that lack AAVR2 binding can be bioengineered to engage with AAVR2. Finally, we overexpressed a minimal functional AAVR2 to enhance AAV transduction in vivo. Our study provides insights into AAV biology and clinically deployable solutions to reduce dose-related toxicities associated with AAV vectors.

Keywords: AAVR2; CRISPRa; KIAA0319L; capsid; carboxypeptidase D; gene transfer; genetic therapy; genome-wide CRISPR screen; molecular therapy; serotypes.

MeSH terms

Animals
Capsid / metabolism
Capsid Proteins / metabolism
Cryoelectron Microscopy
Dependovirus* / genetics
Dependovirus* / metabolism
Genetic Therapy / methods
Genetic Vectors / genetics
Genetic Vectors / metabolism
HEK293 Cells
Humans
Mice
Protein Binding
Receptors, Virus* / chemistry
Receptors, Virus* / metabolism
Transduction, Genetic

Substances

Receptors, Virus
Capsid Proteins