title: 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.
abstract: BACKGROUND
  For 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.
  
  
  METHODS
  To 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.
  
  
  RESULTS
  Transduction 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.
  
  
  CONCLUSIONS
  Using 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.
authors.affiliation: Department G402, Pharmacology of Cancer Treatment, German Cancer Research Center, INF 280, D-69120, Heidelberg, Germany. mstiefelhagen@gmail.com
authors.forename: Marius
authors.initials: M
authors.lastname: Stiefelhagen
authors.forename: Leopold
authors.initials: L
authors.lastname: Sellner
authors.forename: Jürgen A
authors.initials: JA
authors.lastname: Kleinschmidt
authors.forename: Anna
authors.initials: A
authors.lastname: Jauch
authors.forename: Stephanie
authors.initials: S
authors.lastname: Laufs
authors.forename: Frederik
authors.initials: F
authors.lastname: Wenz
authors.forename: W Jens
authors.initials: WJ
authors.lastname: Zeller
authors.forename: Stefan
authors.initials: S
authors.lastname: Fruehauf
authors.forename: Marlon R
authors.initials: MR
authors.lastname: Veldwijk
references.citation: Science. 2001 Aug 3;293(5531):876-80
references.pmid: 11423618
references.citation: Nat Biotechnol. 2003 Sep;21(9):1040-6
references.pmid: 12897791
references.citation: Nat Med. 1999 Jan;5(1):78-82
references.pmid: 9883843
references.citation: J Virol. 1997 Nov;71(11):8262-7
references.pmid: 9343178
references.citation: Blood. 1997 Mar 15;89(6):2167-75
references.pmid: 9058741
references.citation: Blood. 1999 Mar 15;93(6):1882-94
references.pmid: 10068661
references.citation: Nat Med. 1999 Jan;5(1):71-7
references.pmid: 9883842
references.citation: Mol Ther. 2003 Jul;8(1):151-7
references.pmid: 12842438
references.citation: Mol Ther. 2001 Sep;4(3):174-81
references.pmid: 11545607
references.citation: Hum Gene Ther. 2005 Jul;16(7):792-8
references.pmid: 16000061
references.citation: Pediatr Res. 2005 Dec;58(6):1143-7
references.pmid: 16306183
references.citation: Hum Gene Ther. 1997 Nov 1;8(16):1911-20
references.pmid: 9382957
references.citation: Hum Gene Ther. 2004 Dec;15(12):1207-18
references.pmid: 15684697
references.citation: Curr Gene Ther. 2005 Jun;5(3):311-21
references.pmid: 15975008
references.citation: Hum Gene Ther. 2003 Jul 20;14(11):1079-88
references.pmid: 12885347
references.citation: Hum Gene Ther. 1998 Mar 20;9(5):695-706
references.pmid: 9551617
references.citation: Blood. 2002 May 1;99(9):3472-5
references.pmid: 11964322
references.citation: Cancer Cell. 2002 Aug;2(2):117-25
references.pmid: 12204532
references.citation: Nat Genet. 1996 Apr;12(4):368-75
references.pmid: 8630489
references.citation: N Engl J Med. 1999 Jul 15;341(3):164-72
references.pmid: 10403855
references.citation: Natl Med J India. 2006 Sep-Oct;19(5):255-63
references.pmid: 17203680
references.citation: Gene Ther. 2000 Feb;7(3):183-95
references.pmid: 10694794
references.citation: J Virol. 1998 Feb;72(2):1438-45
references.pmid: 9445046
references.citation: J Virol. 2005 Jan;79(1):609-14
references.pmid: 15596854
references.citation: Chromosome Res. 2002;10(3):223-32
references.pmid: 12067211
references.citation: J Virol. 1996 May;70(5):3227-34
references.pmid: 8627803
references.citation: Gene Ther. 2003 Jun;10(11):955-63
references.pmid: 12756416
references.citation: Radiother Oncol. 2004 Sep;72(3):341-50
references.pmid: 15450734
references.citation: Eur J Cancer. 1999 Jul;35(7):1136-42
references.pmid: 10533460
references.citation: Mol Ther. 2002 Aug;6(2):272-8
references.pmid: 12349826
references.citation: Blood. 2003 Apr 15;101(8):2963-72
references.pmid: 12515715
references.citation: Lung Cancer. 2004 Nov;46(2):179-86
references.pmid: 15474666
references.citation: Nature. 2000 Jul 6;406(6791):82-6
references.pmid: 10894546
references.citation: Blood. 2004 Feb 15;103(4):1253-60
references.pmid: 14551134
references.citation: Mol Ther. 2001 Jun;3(6):964-75
references.pmid: 11407911
references.citation: Gene Ther. 2003 Jun;10(11):964-76
references.pmid: 12756417
references.citation: Cancer Gene Ther. 2000 Apr;7(4):597-604
references.pmid: 10811478
references.citation: J Virol. 1997 Mar;71(3):1776-83
references.pmid: 9032306
references.citation: Cancer Gene Ther. 2004 Aug;11(8):577-84
references.pmid: 15280909
references.citation: Nat Med. 1996 May;2(5):561-6
references.pmid: 8616716
references.citation: Nat Genet. 2000 Mar;24(3):257-61
references.pmid: 10700178
references.citation: Hum Gene Ther. 2005 Jun;16(6):699-710
references.pmid: 15960601
references.citation: Gene Ther. 1999 Jun;6(6):973-85
references.pmid: 10455399
references.citation: Science. 2003 Oct 17;302(5644):415-9
references.pmid: 14564000
references.citation: Blood. 2004 Dec 1;104(12):3445-53
references.pmid: 15292064