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Mol Ther. 2016 Jun;24(6):1050-1061. doi: 10.1038/mt.2016.62. Epub 2016 Mar 28.

Pulmonary Targeting of Adeno-associated Viral Vectors by Next-generation Sequencing-guided Screening of Random Capsid Displayed Peptide Libraries.

Author information

1
Department of Oncology and Hematology, University Medical Center Hamburg-Eppendorf, Hubertus Wald Cancer Center, Hamburg, Germany.
2
Section of Experimental Pneumology, Research Center Borstel, Airway Research Center North, Borstel, Germany.
3
Heinrich-Pette-Institute, Leibniz Institute for Experimental Virology, AG96 Technology-Platform, Hamburg, Germany.
4
University Hospital Heidelberg, Heidelberg, and DZHK (German Center for Cardiovascular Research), partner site Heidelberg/Mannheim, Heidelberg, Germany.
5
Department of Internal Medicine, Division of Molecular Medicine, University of New Mexico Cancer Center, University of New Mexico School of Medicine, Albuquerque, New Mexico, USA.
6
Department of Internal Medicine, Division of Hematology/Oncology, University of New Mexico Cancer Center, University of New Mexico School of Medicine, Albuquerque, New Mexico, USA.
7
Deutsches Krebsforschungszentrum, Heidelberg, Germany.
8
Department of Oncology and Hematology, University Medical Center Hamburg-Eppendorf, Hubertus Wald Cancer Center, Hamburg, Germany; Current address: Department of Hematology and Oncology, Augsburg Medical Center, Augsburg, Germany. Electronic address: m.trepel@uke.de.

Abstract

Vectors mediating strong, durable, and tissue-specific transgene expression are mandatory for safe and effective gene therapy. In settings requiring systemic vector administration, the availability of suited vectors is extremely limited. Here, we present a strategy to select vectors with true specificity for a target tissue from random peptide libraries displayed on adeno-associated virus (AAV) by screening the library under circulation conditions in a murine model. Guiding the in vivo screening by next-generation sequencing, we were able to monitor the selection kinetics and to determine the right time point to discontinue the screening process. The establishment of different rating scores enabled us to identify the most specifically enriched AAV capsid candidates. As proof of concept, a capsid variant was selected that specifically and very efficiently delivers genes to the endothelium of the pulmonary vasculature after intravenous administration. This technical approach of selecting target-specific vectors in vivo is applicable to any given tissue of interest and therefore has broad implications in translational research and medicine.

PMID:
27018516
PMCID:
PMC4923327
DOI:
10.1038/mt.2016.62
[Indexed for MEDLINE]
Free PMC Article

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