Application of a haematopoetic progenitor cell-targeted adeno-associated viral AAV vector established by selection of an AAV random peptide library on a leukaemia cell lineReport as inadecuate




Application of a haematopoetic progenitor cell-targeted adeno-associated viral AAV vector established by selection of an AAV random peptide library on a leukaemia cell line - Download this document for free, or read online. Document in PDF available to download.

Genetic Vaccines and Therapy

, 6:12

First Online: 12 September 2008Received: 28 March 2008Accepted: 12 September 2008

Abstract

BackgroundFor many promising target cells e.g.: haematopoeitic progenitors, the susceptibility to standard adeno-associated viral AAV vectors is low. Advancements in vector development now allows the generation of target cell-selected AAV capsid mutants.

MethodsTo determine its suitability, the method was applied on a chronic myelogenous leukaemia CML cell line K562 to obtain a CML-targeted vector and the resulting vectors tested on leukaemia, non-leukaemia, primary human CML and CD34 peripheral blood progenitor cells PBPC; standard AAV2 and a random capsid mutant vector served as controls.

ResultsTransduction of CML BV173, EM3, K562 and Lama84 and AML HL60 and KG1a cell lines with the capsid mutants resulted in an up to 36-fold increase in CML transduction efficiency K562: 2-fold, 60% ± 2% green fluorescent protein GFP cells; BV173: 9-fold, 37% ± 2% GFP cells; Lama84: 36-fold, 29% ± 2% GFP cells compared to controls. For AML KG1a, HL60 and one CML cell line EM3, no significant transduction <1% GFP cells was observed for any vector. Although the capsid mutant clone was established on a cell line, proof-of-principle experiments using primary human cells were performed. For CML 3.2-fold, mutant: 1.75% ± 0.45% GFP cells, p = 0.03 and PBPC 3.5-fold, mutant: 4.21% ± 3.40% GFP cells a moderate increase in gene transfer of the capsid mutant compared to control vectors was observed.

ConclusionUsing an AAV random peptide library on a CML cell line, we were able to generate a capsid mutant, which transduced CML cell lines and primary human haematopoietic progenitor cells with higher efficiency than standard recombinant AAV vectors.

Electronic supplementary materialThe online version of this article doi:10.1186-1479-0556-6-12 contains supplementary material, which is available to authorized users.

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Author: Marius Stiefelhagen - Leopold Sellner - Jürgen A Kleinschmidt - Anna Jauch - Stephanie Laufs - Frederik Wenz - W Jens Ze

Source: https://link.springer.com/







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