Journal article
GPR108 Is a Highly Conserved AAV Entry Factor
Molecular therapy, Vol.28(2), pp.367-381
02/05/2020
DOI: 10.1016/j.ymthe.2019.11.005
PMCID: PMC7000996
PMID: 31784416
Abstract
Adeno-associated virus (AAV) is a highly promising gene transfer vector, yet major cellular requirements for AAV entry are poorly understood. Using a genome-wide CRISPR screen for entry of evolutionarily divergent serotype AAVrh32.33, we identified GPR108, a member of the G protein-coupled receptor superfamily, as an AAV entry factor. Of greater than 20 divergent AAVs across all AAV clades tested in human cell lines, only AAV5 transduction was unaffected in the GPR108 knockout (KO). GPR108 dependency was further shown in murine and primary cells in vitro. These findings are further validated in vivo, as the Gpr108 KO mouse demonstrates 10- to 100-fold reduced expression for AAV8 and rh32.33 but not AAV5. Mechanistically, both GPR108 N- and C-terminal domains are required for transduction, and on the capsid, a VP1 unique domain that is not conserved on AAV5 can be transferred to confer GPR108 independence onto AAV2 chimeras. In vitro binding and fractionation studies indicate reduced nuclear import and cytosolic accumulation in the absence of GPR108. We thus have identified the second of two AAV entry factors that is conserved between mice and humans relevant both in vitro and in vivo, further providing a mechanistic understanding to the tropism of AAV gene therapy vectors.
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Adeno-associated virus (AAV) has been repurposed as a highly effective gene therapy vector, yet conserved cellular factors required for cellular entry are poorly understood. In this issue of Molecular Therapy, Vandenberghe and colleagues describe identification of a novel conserved AAV entry factor that is required in vivo for AAV transduction.
Details
- Title: Subtitle
- GPR108 Is a Highly Conserved AAV Entry Factor
- Creators
- Amanda M. Dudek - Massachusetts Eye and Ear InfirmaryNerea Zabaleta - Massachusetts Eye and Ear InfirmaryEric Zinn - Massachusetts Eye and Ear InfirmarySirika Pillay - Stanford University School of MedicineJames Zengel - Stanford University School of MedicineCaryn Porter - Massachusetts General HospitalJennifer Santos Franceschini - Massachusetts Eye and Ear InfirmaryReynette Estelien - Massachusetts Eye and Ear InfirmaryJan E Carette - Stanford University School of MedicineGuo Ling Zhou - Massachusetts General HospitalLuk H. Vandenberghe - Massachusetts Eye and Ear Infirmary
- Resource Type
- Journal article
- Publication Details
- Molecular therapy, Vol.28(2), pp.367-381
- DOI
- 10.1016/j.ymthe.2019.11.005
- PMID
- 31784416
- PMCID
- PMC7000996
- NLM abbreviation
- Mol Ther
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Publisher
- Elsevier Inc
- Number of pages
- 15
- Language
- English
- Date published
- 02/05/2020
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984822990102771
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