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Correction of murine Rag2 severe combined immunodeficiency by lentiviral gene therapy using a codon-optimized RAG2 therapeutic transgene.
Mol Ther. 2012 Oct; 20(10):1968-80.MT

Abstract

Recombination activating gene 2 (RAG2) deficiency results in severe combined immunodeficiency (SCID) with complete lack of T and B lymphocytes. Initial gammaretroviral gene therapy trials for other types of SCID proved effective, but also revealed the necessity of safe vector design. We report the development of lentiviral vectors with the spleen focus forming virus (SF) promoter driving codon-optimized human RAG2 (RAG2co), which improved phenotype amelioration compared to native RAG2 in Rag2(-/-) mice. With the RAG2co therapeutic transgene, T-cell receptor (TCR) and immunoglobulin repertoire, T-cell mitogen responses, plasma immunoglobulin levels and T-cell dependent and independent specific antibody responses were restored. However, the thymus double positive T-cell population remained subnormal, possibly due to the SF virus derived element being sensitive to methylation/silencing in the thymus, which was prevented by replacing the SF promoter by the previously reported silencing resistant element (ubiquitous chromatin opening element (UCOE)), and also improved B-cell reconstitution to eventually near normal levels. Weak cellular promoters were effective in T-cell reconstitution, but deficient in B-cell reconstitution. We conclude that immune functions are corrected in Rag2(-/-) mice by genetic modification of stem cells using the UCOE driven codon-optimized RAG2, providing a valid optional vector for clinical implementation.

Authors+Show Affiliations

Department of Hematology, Erasmus University Medical Center, Rotterdam, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22692499

Citation

van Til, Niek P., et al. "Correction of Murine Rag2 Severe Combined Immunodeficiency By Lentiviral Gene Therapy Using a Codon-optimized RAG2 Therapeutic Transgene." Molecular Therapy : the Journal of the American Society of Gene Therapy, vol. 20, no. 10, 2012, pp. 1968-80.
van Til NP, de Boer H, Mashamba N, et al. Correction of murine Rag2 severe combined immunodeficiency by lentiviral gene therapy using a codon-optimized RAG2 therapeutic transgene. Mol Ther. 2012;20(10):1968-80.
van Til, N. P., de Boer, H., Mashamba, N., Wabik, A., Huston, M., Visser, T. P., Fontana, E., Poliani, P. L., Cassani, B., Zhang, F., Thrasher, A. J., Villa, A., & Wagemaker, G. (2012). Correction of murine Rag2 severe combined immunodeficiency by lentiviral gene therapy using a codon-optimized RAG2 therapeutic transgene. Molecular Therapy : the Journal of the American Society of Gene Therapy, 20(10), 1968-80. https://doi.org/10.1038/mt.2012.110
van Til NP, et al. Correction of Murine Rag2 Severe Combined Immunodeficiency By Lentiviral Gene Therapy Using a Codon-optimized RAG2 Therapeutic Transgene. Mol Ther. 2012;20(10):1968-80. PubMed PMID: 22692499.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Correction of murine Rag2 severe combined immunodeficiency by lentiviral gene therapy using a codon-optimized RAG2 therapeutic transgene. AU - van Til,Niek P, AU - de Boer,Helen, AU - Mashamba,Nomusa, AU - Wabik,Agnieszka, AU - Huston,Marshall, AU - Visser,Trudi P, AU - Fontana,Elena, AU - Poliani,Pietro Luigi, AU - Cassani,Barbara, AU - Zhang,Fang, AU - Thrasher,Adrian J, AU - Villa,Anna, AU - Wagemaker,Gerard, Y1 - 2012/06/12/ PY - 2012/6/14/entrez PY - 2012/6/14/pubmed PY - 2013/3/30/medline SP - 1968 EP - 80 JF - Molecular therapy : the journal of the American Society of Gene Therapy JO - Mol. Ther. VL - 20 IS - 10 N2 - Recombination activating gene 2 (RAG2) deficiency results in severe combined immunodeficiency (SCID) with complete lack of T and B lymphocytes. Initial gammaretroviral gene therapy trials for other types of SCID proved effective, but also revealed the necessity of safe vector design. We report the development of lentiviral vectors with the spleen focus forming virus (SF) promoter driving codon-optimized human RAG2 (RAG2co), which improved phenotype amelioration compared to native RAG2 in Rag2(-/-) mice. With the RAG2co therapeutic transgene, T-cell receptor (TCR) and immunoglobulin repertoire, T-cell mitogen responses, plasma immunoglobulin levels and T-cell dependent and independent specific antibody responses were restored. However, the thymus double positive T-cell population remained subnormal, possibly due to the SF virus derived element being sensitive to methylation/silencing in the thymus, which was prevented by replacing the SF promoter by the previously reported silencing resistant element (ubiquitous chromatin opening element (UCOE)), and also improved B-cell reconstitution to eventually near normal levels. Weak cellular promoters were effective in T-cell reconstitution, but deficient in B-cell reconstitution. We conclude that immune functions are corrected in Rag2(-/-) mice by genetic modification of stem cells using the UCOE driven codon-optimized RAG2, providing a valid optional vector for clinical implementation. SN - 1525-0024 UR - https://www.unboundmedicine.com/medline/citation/22692499/Correction_of_murine_Rag2_severe_combined_immunodeficiency_by_lentiviral_gene_therapy_using_a_codon_optimized_RAG2_therapeutic_transgene_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1525-0016(16)32193-1 DB - PRIME DP - Unbound Medicine ER -