Endothelial cells (EC) are targets in gene therapy and regenerative medicine, but they are inefficiently transduced with adeno-associated virus (AAV) vectors of various serotypes. To identify barriers hampering efficient transduction and to develop an optimized AAV variant for EC transduction, we screened an AAV serotype 2-based peptide display library on primary human macrovascular EC. Using a new high-throughput selection and monitoring protocol, we identified a capsid variant, AAV-V EC, which outperformed the parental serotype as well as first-generation targeting vectors in EC transduction. AAV vector uptake was improved, resulting in significantly higher transgene expression levels from single-stranded vector genomes detectable within a few hours post-transduction. Notably, AAV-V EC transduced not only proliferating EC but also quiescent EC, although higher particle-per-cell ratios had to be applied. Also, induced pluripotent stem cell-derived endothelial progenitor cells, a novel tool in regenerative medicine and gene therapy, were highly susceptible toward AAV-V EC transduction. Thus, overcoming barriers by capsid engineering significantly expands the AAV tool kit for a wide range of applications targeting EC.
|Number of pages||13|
|Journal||Human Gene Therapy|
|Early online date||13 Aug 2019|
|Publication status||Published (in print/issue) - 23 Sept 2019|
Bibliographical noteFunding Information:
This work was supported by grants from University Hospital Cologne (Kln Fortune) to H.B. and M.O., the Center for Molecular Medicine Cologne (CMMC) to H.B., BMBF and MWK Lower Saxony-funded Professorinnenprogramm Niedersachsen to H.B., the DFG-funded KFO286 to H.B., the DFG-funded cluster of excellence REBIRTH EXC62/2 to A.S. and H.B., and the DFG-funded SFB738 to A.S. L.Z. held a scholarship from the German Academic Exchange Service (DAAD).
© 2019, Mary Ann Liebert, Inc.
Copyright 2019 Elsevier B.V., All rights reserved.
- AAV vectors
- endothelial cells
- induced pluripotent stem cells