title: Preclinical demonstration of lentiviral vector-mediated correction of immunological and metabolic abnormalities in models of adenosine deaminase deficiency.
abstract: Gene transfer into autologous hematopoietic stem cells by γ-retroviral vectors (gRV) is an effective treatment for adenosine deaminase (ADA)-deficient severe combined immunodeficiency (SCID). However, current gRV have significant potential for insertional mutagenesis as reported in clinical trials for other primary immunodeficiencies. To improve the efficacy and safety of ADA-SCID gene therapy (GT), we generated a self-inactivating lentiviral vector (LV) with a codon-optimized human cADA gene under the control of the short form elongation factor-1α promoter (LV EFS ADA). In ADA(-/-) mice, LV EFS ADA displayed high-efficiency gene transfer and sufficient ADA expression to rescue ADA(-/-) mice from their lethal phenotype with good thymic and peripheral T- and B-cell reconstitution. Human ADA-deficient CD34(+) cells transduced with 1-5 × 10(7) TU/ml had 1-3 vector copies/cell and expressed 1-2x of normal endogenous levels of ADA, as assayed in vitro and by transplantation into immune-deficient mice. Importantly, in vitro immortalization assays demonstrated that LV EFS ADA had significantly less transformation potential compared to gRV vectors, and vector integration-site analysis by nrLAM-PCR of transduced human cells grown in immune-deficient mice showed no evidence of clonal skewing. These data demonstrated that the LV EFS ADA vector can effectively transfer the human ADA cDNA and promote immune and metabolic recovery, while reducing the potential for vector-mediated insertional mutagenesis.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Denise A
authors.initials: DA
authors.lastname: Carbonaro
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Jin
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.
authors.forename: Claudia
authors.initials: C
authors.lastname: Montiel-Equihua
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Sabine
authors.initials: S
authors.lastname: Geiger
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.
authors.forename: Marlene
authors.initials: M
authors.lastname: Carmo
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Cooper
authors.affiliation: Purine Research Laboratory, St Thomas Hospital, London, UK.
authors.forename: Lynette
authors.initials: L
authors.lastname: Fairbanks
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Kaufman
authors.affiliation: Department of Paediatric Pathology, Great Ormond Street Hospital, London, UK.
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Sebire
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Roger P
authors.initials: RP
authors.lastname: Hollis
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Blundell
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Shantha
authors.initials: S
authors.lastname: Senadheera
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Fu
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Arineh
authors.initials: A
authors.lastname: Sahaghian
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.
authors.forename: Rebecca Y
authors.initials: RY
authors.lastname: Chan
authors.affiliation: Department of Medicine Statistics Core, University of California, Los Angeles, California, USA.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Cornetta
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA; Department of Pediatrics (Hematology/Oncology), Mattel Children's Hospital, University of California, Los Angeles, California, USA.
authors.forename: Donald B
authors.initials: DB
authors.lastname: Kohn
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK. Electronic address: h.gaspar@ucl.ac.uk.
authors.forename: H Bobby
authors.initials: HB
authors.lastname: Gaspar
mesh_terms: D000243:Adenosine Deaminase / Q000172:deficiency* / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D000361:Agammaglobulinemia / Q000235:genetics / Q000276:immunology* / Q000473:pathology / Q000628:therapy*; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005260:Female; D005822:Genetic Vectors / Q000009:adverse effects*; D057809:HEK293 Cells; D019073:HT29 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D020648:Peptide Elongation Factor 1 / Q000235:genetics*; D011401:Promoter Regions, Genetic; D016511:Severe Combined Immunodeficiency / Q000235:genetics / Q000276:immunology* / Q000473:pathology / Q000628:therapy*; D013601:T-Lymphocytes / Q000276:immunology; D014161:Transduction, Genetic; D016662:Virus Integration
chemical_list: D020648:Peptide Elongation Factor 1; D000243:Adenosine Deaminase