Journal article
Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses
Molecular therapy. Methods & clinical development, Vol.12, pp.202-222
01/18/2019
DOI: 10.1016/j.omtm.2019.01.003
PMCID: PMC6360332
PMID: 30766894
Abstract
Parvoviruses are highly attractive templates for the engineering of safe, efficient, and specific gene therapy vectors, as best exemplified by adeno-associated virus (AAV). Another candidate that currently garners increasing attention is human bocavirus 1 (HBoV1). Notably, HBoV1 capsids can cross-package recombinant (r)AAV2 genomes, yielding rAAV2/HBoV1 chimeras that specifically transduce polarized human airway epithelia (pHAEs). Here, we largely expanded the repertoire of rAAV/BoV chimeras, by assembling packaging plasmids encoding the capsid genes of four additional primate bocaviruses, HBoV2–4 and GBoV (Gorilla BoV). Capsid protein expression and efficient rAAV cross-packaging were validated by immunoblotting and qPCR, respectively. Interestingly, not only HBoV1 but also HBoV4 and GBoV transduced pHAEs as well as primary human lung organoids. Flow cytometry analysis of pHAEs revealed distinct cellular specificities between the BoV isolates, with HBoV1 targeting ciliated, club, and KRT5+ basal cells, whereas HBoV4 showed a preference for KRT5+ basal cells. Surprisingly, primary human hepatocytes, skeletal muscle cells, and T cells were also highly amenable to rAAV/BoV transduction. Finally, we adapted our pipeline for AAV capsid gene shuffling to all five BoV isolates. Collectively, our chimeric rAAV/BoV vectors and bocaviral capsid library represent valuable new resources to dissect BoV biology and to breed unique gene therapy vectors.
Details
- Title: Subtitle
- Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses
- Creators
- Julia Fakhiri - Heidelberg UniversityMarc A. Schneider - University Hospital HeidelbergJens Puschhof - Royal Netherlands Academy of Arts and SciencesMegan Stanifer - University Hospital HeidelbergVerena Schildgen - Kliniken der Stadt KölnStefan Holderbach - Heidelberg UniversityYannik Voss - Heidelberg UniversityJihad El Andari - Heidelberg UniversityOliver Schildgen - Kliniken der Stadt KölnSteeve Boulant - University Hospital HeidelbergMichael Meister - University Hospital HeidelbergHans Clevers - Royal Netherlands Academy of Arts and SciencesZiying Yan - University of IowaJianming Qiu - University of Kansas Medical CenterDirk Grimm - Heidelberg University
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Methods & clinical development, Vol.12, pp.202-222
- Publisher
- American Society of Gene & Cell Therapy
- DOI
- 10.1016/j.omtm.2019.01.003
- PMID
- 30766894
- PMCID
- PMC6360332
- ISSN
- 2329-0501
- eISSN
- 2329-0501
- Language
- English
- Date published
- 01/18/2019
- Academic Unit
- Anatomy and Cell Biology
- Record Identifier
- 9984284350602771
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