title: Genotoxic potential of lineage-specific lentivirus vectors carrying the beta-globin locus control region.
abstract: Insertional mutagenesis by long terminal repeat (LTR) enhancers in gamma-retrovirus-based vectors (GVs) in clinical trials has prompted deeper investigations into vector genotoxicity. Experimentally, self-inactivating (SIN) lentivirus vectors (LVs) and GV containing internal promoters/enhancers show reduced genotoxicity, although strong ubiquitously-active enhancers dysregulate genes independent of vector type/design. Herein, we explored the genotoxicity of beta-globin (BG) locus control region (LCR), a strong long-range lineage-specific-enhancer, with/without insulator (Ins) elements in LV using primary hematopoietic progenitors to generate in vitro immortalization (IVIM) assay mutants. LCR-containing LV had approximately 200-fold lower transforming potential, compared to the conventional GV. The LCR perturbed expression of few genes in a 300 kilobase (kb) proviral vicinity but no upregulation of genes associated with cancer, including an erythroid-specific transcription factor occurred. A further twofold reduction in transforming activity was observed with insulated LCR-containing LV. Our data indicate that toxicology studies of LCR-containing LV in mice will likely not yield any insertional oncogenesis with the numbers of animals that can be practically studied.
authors.affiliation: Division of Experimental Hematology/Cancer Biology, Cincinnati Children's Research Foundation, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229, USA.
authors.forename: Paritha I
authors.initials: PI
authors.lastname: Arumugam
authors.forename: Tomoyasu
authors.initials: T
authors.lastname: Higashimoto
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Urbinati
authors.forename: Ute
authors.initials: U
authors.lastname: Modlich
authors.forename: Shawna
authors.initials: S
authors.lastname: Nestheide
authors.forename: Ping
authors.initials: P
authors.lastname: Xia
authors.forename: Catherine
authors.initials: C
authors.lastname: Fox
authors.forename: Andrea
authors.initials: A
authors.lastname: Corsinotti
authors.forename: Christopher
authors.initials: C
authors.lastname: Baum
authors.forename: Punam
authors.initials: P
authors.lastname: Malik
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000009:adverse effects / Q000235:genetics*; D038101:Insulator Elements / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D019901:Locus Control Region / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D016254:Mutagenesis, Insertional; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic; D055544:beta-Globins / Q000235:genetics*
chemical_list: D055544:beta-Globins
references.citation: Mol Ther. 2008 Mar;16(3):534-40
references.pmid: 18209733
references.citation: J Clin Invest. 2004 Oct;114(7):953-62
references.pmid: 15467834
references.citation: Leukemia. 2008 Aug;22(8):1519-28
references.pmid: 18496560
references.citation: Mol Interv. 2004 Dec;4(6):321-2
references.pmid: 15616160
references.citation: Cell Stem Cell. 2009 Feb 6;4(2):129-40
references.pmid: 19200802
references.citation: Nature. 2006 Aug 17;442(7104):818-22
references.pmid: 16862118
references.citation: Blood. 2007 Sep 15;110(6):1770-8
references.pmid: 17507662
references.citation: Blood. 2004 Dec 1;104(12):3445-53
references.pmid: 15292064
references.citation: J Clin Invest. 2004 Oct;114(7):889-91
references.pmid: 15467825
references.citation: Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10618-22
references.pmid: 1438257
references.citation: Mol Cell Endocrinol. 2003 Dec 31;213(1):23-30
references.pmid: 15062571
references.citation: Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1457-62
references.pmid: 16432223
references.citation: Mol Ther. 2006 Feb;13(2):391-400
references.pmid: 16226060
references.citation: J Clin Invest. 2008 Sep;118(9):3132-42
references.pmid: 18688285
references.citation: Nat Med. 2006 Apr;12(4):401-9
references.pmid: 16582916
references.citation: Cell. 1987 Dec 24;51(6):975-85
references.pmid: 3690667
references.citation: Nature. 2006 Sep 21;443(7109):E5; discussion E6-7
references.pmid: 16988660
references.citation: Mol Ther. 2009 May;17(5):851-6
references.pmid: 19293778
references.citation: Int Rev Cytol. 2005;246:31-58
references.pmid: 16164966
references.citation: Int J Oncol. 2008 Jul;33(1):17-24
references.pmid: 18575746
references.citation: PLoS One. 2009;4(2):e4571
references.pmid: 19238208
references.citation: J Clin Invest. 2007 Aug;117(8):2241-9
references.pmid: 17671654
references.citation: Mol Ther. 2008 Mar;16(3):439-49
references.pmid: 18227842
references.citation: Blood. 2006 Oct 15;108(8):2545-53
references.pmid: 16825499
references.citation: Blood. 2005 Oct 1;106(7):2530-3
references.pmid: 15933056
references.citation: Blood. 2002 Nov 1;100(9):3077-86
references.pmid: 12384402
references.citation: J Clin Invest. 2008 Sep;118(9):3143-50
references.pmid: 18688286
references.citation: Mol Ther. 2008 Mar;16(3):525-33
references.pmid: 18195719
references.citation: Genes Cells. 2003 Jun;8(6):537-46
references.pmid: 12786944
references.citation: Nat Biotechnol. 2006 Jun;24(6):687-96
references.pmid: 16732270
references.citation: Mol Ther. 2009 Sep;17(9):1527-36
references.pmid: 19384292
references.citation: Mol Ther. 2007 Oct;15(10):1863-71
references.pmid: 17622240
references.citation: Eur J Biochem. 1996 May 1;237(3):833-40
references.pmid: 8647132
references.citation: Proc Natl Acad Sci U S A. 2002 May 14;99(10):6883-8
references.pmid: 12011446
references.citation: Nature. 2006 Apr 27;440(7088):1123
references.pmid: 16641981
references.citation: Mol Ther. 2008 Apr;16(4):718-25
references.pmid: 18334985
references.citation: Mol Ther. 2009 Apr;17(4):716-24
references.pmid: 19240697
references.citation: Cell. 1999 Aug 6;98(3):387-96
references.pmid: 10458613
references.citation: Nat Rev Microbiol. 2005 Nov;3(11):848-58
references.pmid: 16175173
references.citation: J Clin Invest. 2009 Apr;119(4):964-75
references.pmid: 19307726
references.citation: Blood. 2008 Feb 15;111(4):1866-75
references.pmid: 17991809