Successful Preclinical Development of Gene Therapy for Recombinase-Activating Gene-1-Deficient SCID
Laura Garcia-Perez, Marja van Eggermond, Lieke van Roon, Sandra A Vloemans, Martijn Cordes, Axel Schambach, Michael Rothe, Dagmar Berghuis, Chantal Lagresle-Peyrou, Marina Cavazzana, Fang Zhang, Adrian J Thrasher, Daniela Salvatori, Pauline Meij, Anna Villa, Jacques J M Van Dongen, Jaap-Jan Zwaginga, Mirjam van der Burg, H Bobby Gaspar, Arjan Lankester, Frank J T Staal, Karin Pike-Overzet, Laura Garcia-Perez, Marja van Eggermond, Lieke van Roon, Sandra A Vloemans, Martijn Cordes, Axel Schambach, Michael Rothe, Dagmar Berghuis, Chantal Lagresle-Peyrou, Marina Cavazzana, Fang Zhang, Adrian J Thrasher, Daniela Salvatori, Pauline Meij, Anna Villa, Jacques J M Van Dongen, Jaap-Jan Zwaginga, Mirjam van der Burg, H Bobby Gaspar, Arjan Lankester, Frank J T Staal, Karin Pike-Overzet
Abstract
Recombinase-activating gene-1 (RAG1)-deficient severe combined immunodeficiency (SCID) patients lack B and T lymphocytes due to the inability to rearrange immunoglobulin and T cell receptor genes. Gene therapy is an alternative for those RAG1-SCID patients who lack a suitable bone marrow donor. We designed lentiviral vectors with different internal promoters driving codon-optimized RAG1 to ensure optimal expression. We used Rag1 -/- mice as a preclinical model for RAG1-SCID to assess the efficacy of the various vectors. We observed that B and T cell reconstitution directly correlated with RAG1 expression. Mice with low RAG1 expression showed poor immune reconstitution; however, higher expression resulted in phenotypic and functional lymphocyte reconstitution comparable to mice receiving wild-type stem cells. No signs of genotoxicity were found. Additionally, RAG1-SCID patient CD34+ cells transduced with our clinical RAG1 vector and transplanted into NSG mice led to improved human B and T cell development. Considering this efficacy outcome, together with favorable safety data, these results substantiate the need for a clinical trial for RAG1-SCID.
Keywords: B lymphocytes; CD34+ cells; RAG1; SCID; T lymphocytes; gene rearrangement; gene therapy; lentiviral vector.
© 2020 The Authors.
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References
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