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Preclinical demonstration of lentiviral vector-mediated correction of immunological and metabolic abnormalities in models of adenosine deaminase deficiency.
Mol Ther. 2014 Mar; 22(3):607-622.MT

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+Show Affiliations

Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Purine Research Laboratory, St Thomas Hospital, London, UK.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Department of Paediatric Pathology, Great Ormond Street Hospital, London, UK.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA.Department of Medicine Statistics Core, University of California, Los Angeles, California, USA.Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana, USA.Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.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.Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK. Electronic address: h.gaspar@ucl.ac.uk.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

24256635

Citation

Carbonaro, Denise A., et al. "Preclinical Demonstration of Lentiviral Vector-mediated Correction of Immunological and Metabolic Abnormalities in Models of Adenosine Deaminase Deficiency." Molecular Therapy : the Journal of the American Society of Gene Therapy, vol. 22, no. 3, 2014, pp. 607-622.
Carbonaro DA, Zhang L, Jin X, et al. Preclinical demonstration of lentiviral vector-mediated correction of immunological and metabolic abnormalities in models of adenosine deaminase deficiency. Mol Ther. 2014;22(3):607-622.
Carbonaro, D. A., Zhang, L., Jin, X., Montiel-Equihua, C., Geiger, S., Carmo, M., Cooper, A., Fairbanks, L., Kaufman, M. L., Sebire, N. J., Hollis, R. P., Blundell, M. P., Senadheera, S., Fu, P. Y., Sahaghian, A., Chan, R. Y., Wang, X., Cornetta, K., Thrasher, A. J., ... Gaspar, H. B. (2014). Preclinical demonstration of lentiviral vector-mediated correction of immunological and metabolic abnormalities in models of adenosine deaminase deficiency. Molecular Therapy : the Journal of the American Society of Gene Therapy, 22(3), 607-622. https://doi.org/10.1038/mt.2013.265
Carbonaro DA, et al. Preclinical Demonstration of Lentiviral Vector-mediated Correction of Immunological and Metabolic Abnormalities in Models of Adenosine Deaminase Deficiency. Mol Ther. 2014;22(3):607-622. PubMed PMID: 24256635.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Preclinical demonstration of lentiviral vector-mediated correction of immunological and metabolic abnormalities in models of adenosine deaminase deficiency. AU - Carbonaro,Denise A, AU - Zhang,Lin, AU - Jin,Xiangyang, AU - Montiel-Equihua,Claudia, AU - Geiger,Sabine, AU - Carmo,Marlene, AU - Cooper,Aaron, AU - Fairbanks,Lynette, AU - Kaufman,Michael L, AU - Sebire,Neil J, AU - Hollis,Roger P, AU - Blundell,Michael P, AU - Senadheera,Shantha, AU - Fu,Pei-Yu, AU - Sahaghian,Arineh, AU - Chan,Rebecca Y, AU - Wang,Xiaoyan, AU - Cornetta,Kenneth, AU - Thrasher,Adrian J, AU - Kohn,Donald B, AU - Gaspar,H Bobby, Y1 - 2013/11/20/ PY - 2013/10/11/received PY - 2013/11/11/accepted PY - 2013/11/22/entrez PY - 2013/11/22/pubmed PY - 2014/11/5/medline SP - 607 EP - 622 JF - Molecular therapy : the journal of the American Society of Gene Therapy JO - Mol. Ther. VL - 22 IS - 3 N2 - 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. SN - 1525-0024 UR - https://www.unboundmedicine.com/medline/citation/24256635/Preclinical_demonstration_of_lentiviral_vector_mediated_correction_of_immunological_and_metabolic_abnormalities_in_models_of_adenosine_deaminase_deficiency_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1525-0016(16)31185-6 DB - PRIME DP - Unbound Medicine ER -