title: A self-inactivating lentiviral vector for SCID-X1 gene therapy that does not activate LMO2 expression in human T cells.
abstract: To develop safer and more effective vectors for gene therapy of X-linked severe combined immunodeficiency (SCID-X1), we have evaluated new self-inactivating lentiviral vectors based on the HIV virus. The CL20i4-hgamma(c)-Revgen vector contains the entire human common gamma chain (gamma(c)) genomic sequence driven by the gamma(c) promoter. The CL20i4-EF1alpha-hgamma(c)OPT vector uses a promoter fragment from the eukaryotic elongation factor alpha (EF1alpha) gene to express a codon-optimized human gamma(c) cDNA. Both vectors contain a 400-bp insulator fragment from the chicken beta-globin locus within the self-inactivating long-terminal repeat. Transduction of bone marrow cells using either of these vectors restored T, B, and natural killer lymphocyte development and function in a mouse SCID-X1 transplantation model. Transduction of human CD34(+) bone marrow cells from SCID-X1 patients with either vector restored T-cell development in an in vitro assay. In safety studies using a Jurkat LMO2 activation assay, only the CL20i4-EF1alpha-hgamma(c)OPT vector lacked the ability to transactivate LMO2 protein expression, whereas the CL20i4-hgamma(c)-Revgen vector significantly activated LMO2 protein expression. In addition, the CL20i4-EF1alpha-hgamma(c)OPT vector has not caused any tumors in transplanted mice. We conclude that the CL20i4-EF1alpha-hgamma(c)OPT vector may be suitable for testing in a clinical trial based on these preclinical demonstrations of efficacy and safety.
authors.affiliation: Division of Experimental Hematology, Department of Hematology, St Jude Children's Research Hospital, 262 Danny Thomas Pl., Memphis, TN 38105, USA. sheng.zhou@stjude.org
authors.forename: Sheng
authors.initials: S
authors.lastname: Zhou
authors.forename: Disha
authors.initials: D
authors.lastname: Mody
authors.forename: Suk See
authors.initials: SS
authors.lastname: DeRavin
authors.forename: Julia
authors.initials: J
authors.lastname: Hauer
authors.forename: Taihe
authors.initials: T
authors.lastname: Lu
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Ma
authors.forename: Salima
authors.initials: S
authors.lastname: Hacein-Bey Abina
authors.forename: John T
authors.initials: JT
authors.lastname: Gray
authors.forename: Michael R
authors.initials: MR
authors.lastname: Greene
authors.forename: Marina
authors.initials: M
authors.lastname: Cavazzana-Calvo
authors.forename: Harry L
authors.initials: HL
authors.lastname: Malech
authors.forename: Brian P
authors.initials: BP
authors.lastname: Sorrentino
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism; D016026:Bone Marrow Transplantation / Q000379:methods; D002471:Cell Transformation, Neoplastic; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D053631:Interleukin Receptor Common gamma Subunit / Q000235:genetics; D007438:Introns / Q000235:genetics; D019169:Jurkat Cells / Q000378:metabolism; D007694:Killer Cells, Natural / Q000166:cytology / Q000378:metabolism; D060588:LIM Domain Proteins; D016086:Lentivirus / Q000235:genetics*; D008667:Metalloproteins / Q000378:metabolism*; D051379:Mice; D008817:Mice, Mutant Strains; D016513:Mice, SCID; D020648:Peptide Elongation Factor 1 / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D016511:Severe Combined Immunodeficiency / Q000235:genetics* / Q000276:immunology / Q000628:therapy*; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018952:Antigens, CD34; D004268:DNA-Binding Proteins; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; D060588:LIM Domain Proteins; C513188:Lmo2 protein, mouse; D008667:Metalloproteins; D020648:Peptide Elongation Factor 1
references.citation: Mol Ther. 2000 Feb;1(2):145-53
references.pmid: 10933924
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16406-11
references.pmid: 17062750
references.citation: J Clin Invest. 2008 Sep;118(9):3143-50
references.pmid: 18688286
references.citation: Mol Ther. 2008 Mar;16(3):590-8
references.pmid: 18180772
references.citation: Blood. 2007 Jul 1;110(1):67-73
references.pmid: 17369490
references.citation: Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):575-80
references.pmid: 9012826
references.citation: Science. 2003 Oct 17;302(5644):415-9
references.pmid: 14564000
references.citation: Proc Natl Acad Sci U S A. 2006 Aug 1;103(31):11730-5
references.pmid: 16864781
references.citation: Cell. 1993 Apr 9;73(1):147-57
references.pmid: 8462096
references.citation: Hum Gene Ther. 2007 Apr;18(4):333-43
references.pmid: 17411365
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: Blood. 2004 Oct 15;104(8):2281-90
references.pmid: 15198957
references.citation: J Clin Invest. 2007 Jun;117(6):1456-65
references.pmid: 17549248
references.citation: Blood. 2009 Apr 23;113(17):4114-24
references.pmid: 19168787
references.citation: Blood. 2005 Jun 1;105(11):4314-20
references.pmid: 15701724
references.citation: N Engl J Med. 2002 Apr 18;346(16):1185-93
references.pmid: 11961146
references.citation: Lancet. 2004 Dec 18-31;364(9452):2181-7
references.pmid: 15610804
references.citation: Blood. 1996 Nov 15;88(10):3901-9
references.pmid: 8916956
references.citation: J Clin Invest. 2007 Aug;117(8):2241-9
references.pmid: 17671654
references.citation: N Engl J Med. 1999 Feb 18;340(7):508-16
references.pmid: 10021471
references.citation: Blood. 1999 Nov 1;94(9):3027-36
references.pmid: 10556186
references.citation: Blood. 2007 Jul 1;110(1):116-24
references.pmid: 17374739
references.citation: Science. 1999 Jan 29;283(5402):682-6
references.pmid: 9924027
references.citation: Blood. 2006 Oct 15;108(8):2545-53
references.pmid: 16825499
references.citation: Immunity. 2002 Dec;17(6):749-56
references.pmid: 12479821
references.citation: Mol Ther. 2009 Nov;17(11):1910-8
references.pmid: 19638958
references.citation: JAMA. 2006 Feb 1;295(5):508-18
references.pmid: 16449616
references.citation: Immunity. 1995 Mar;2(3):223-38
references.pmid: 7697543
references.citation: Nature. 2000 Jul 6;406(6791):82-6
references.pmid: 10894546
references.citation: Hum Mol Genet. 1993 Aug;2(8):1099-104
references.pmid: 8401490
references.citation: J Pediatr. 1999 Jun;134(6):740-8
references.pmid: 10356144
references.citation: Mol Ther. 2007 Apr;15(4):782-91
references.pmid: 17299409
references.citation: Annu Rev Immunol. 2004;22:625-55
references.pmid: 15032591
references.citation: Science. 2000 Apr 28;288(5466):669-72
references.pmid: 10784449
references.citation: Nat Med. 1996 Aug;2(8):876-82
references.pmid: 8705856
references.citation: Blood. 2009 May 21;113(21):5104-10
references.pmid: 19286997
references.citation: Lancet. 2003 Feb 15;361(9357):553-60
references.pmid: 12598139
references.citation: Blood. 2000 May 15;95(10):3071-7
references.pmid: 10807771
references.citation: Nat Biotechnol. 2006 Jun;24(6):687-96
references.pmid: 16732270
references.citation: Blood. 2004 Jun 1;103(11):4062-9
references.pmid: 14976042
references.citation: Mol Ther. 2008 Apr;16(4):718-25
references.pmid: 18334985
references.citation: Immunol Rev. 2004 Dec;202:67-83
references.pmid: 15546386
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
references.citation: PLoS Biol. 2004 Aug;2(8):E234
references.pmid: 15314653
references.citation: Blood. 2007 Sep 1;110(5):1448-57
references.pmid: 17456723
references.citation: Nature. 1964 Jul 25;203:420-2
references.pmid: 14197398
references.citation: Stem Cells. 2008 Dec;26(12):3257-66
references.pmid: 18787211