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Pike-Overzet, K.; Rodijk, M.; Ng, Y.Y.; Baert, M.R.M.; Lagresle-Peyrou, C.; Schambach, A.; ... ; Staal, F.J.T. (2011)

Correction of murine Rag1 deficiency by self-inactivating lentiviral vector-mediated gene transfer

Article / Letter to editor
Severe combined immunodeficiency (SCID) patients with an inactivating mutation in recombination activation gene 1 (RAG1) lack B and T cells due to the inability to rearrange immunoglobulin (Ig) and T-cell receptor (TCR) genes. Gene therapy is a valid treatment option for RAG-SCID patients, especially for patients lacking a suitable bone marrow donor, but developing such therapy has proven challenging. As a preclinical model for RAG-SCID, we used Rag1-/- mice and lentiviral self-inactivating (SIN) vectors harboring different internal elements to deliver native or codon-optimized human RAG1 sequences. Treatment resulted in the appearance of B and T cells in peripheral blood and developing B and T cells were detected in central lymphoid organs. Serum Ig levels and Ig and TCR V beta gene segment usage was comparable to wildtype (WT) controls, indicating that RAG-mediated rearrangement took place. Remarkably, relatively low frequencies of B cells produced WT levels of serum...Show moreSevere combined immunodeficiency (SCID) patients with an inactivating mutation in recombination activation gene 1 (RAG1) lack B and T cells due to the inability to rearrange immunoglobulin (Ig) and T-cell receptor (TCR) genes. Gene therapy is a valid treatment option for RAG-SCID patients, especially for patients lacking a suitable bone marrow donor, but developing such therapy has proven challenging. As a preclinical model for RAG-SCID, we used Rag1-/- mice and lentiviral self-inactivating (SIN) vectors harboring different internal elements to deliver native or codon-optimized human RAG1 sequences. Treatment resulted in the appearance of B and T cells in peripheral blood and developing B and T cells were detected in central lymphoid organs. Serum Ig levels and Ig and TCR V beta gene segment usage was comparable to wildtype (WT) controls, indicating that RAG-mediated rearrangement took place. Remarkably, relatively low frequencies of B cells produced WT levels of serum immunoglobulins. Upon stimulation of the TCR, corrected spleen cells proliferated and produced cytokines. In vivo challenge resulted in production of antigen-specific antibodies. No leukemia development as consequence of insertional mutagenesis was observed. The functional reconstitution of the B- as well as the T-cell compartment provides proof-of-principle for therapeutic RAG1 gene transfer in Rag1-/- mice using lentiviral SIN vectors. Leukemia (2011) 25, 1471-1483; doi: 10.1038/leu.2011.106; published online 27 May 2011Show less
gene therapy T lymphocyte SCID severe combined immunodeficiency posttranscriptional regulatory element b-cell differentiation retroviral vectors in-vivo hematopoietic progenitors rag-1-deficient mice sustained correction bone-marrow therapy
All authors
Pike-Overzet, K.; Rodijk, M.; Ng, Y.Y.; Baert, M.R.M.; Lagresle-Peyrou, C.; Schambach, A.; Zhang, F.; Hoeben, R.C.; Hacein-Bey-Abina, S.; Lankester, A.C.; Bredius, R.G.M.; Driessen, G.J.A.; Thrasher, A.J.; Baum, C.; Cavazzana-Calvo, M.; Dongen, J.J.M. van; Staal, F.J.T.
Date
2011-09-30
Journal
Leukemia
Volume
25
Issue
9
Pages
1471 - 1483
DOI
doi:10.1038/leu.2011.106