Gene transfer into hematopoietic stem cells by gamma-retroviral vectors (RVs) is an effective treatment for inherited blood disorders, although potentially limited by the risk of insertional mutagenesis. We evaluated the genomic impact of RV integration in T lymphocytes from adenosine deaminase-deficient severe combined immunodeficiency (ADA-SCID) patients 10 to 30 months after infusion of autologous, genetically corrected CD34(+) cells. Expression profiling on ex vivo T-cell bulk population revealed no difference with respect to healthy controls. To assess the effect of vector integration on gene expression at the single-cell level, primary T-cell clones were isolated from 2 patients. T-cell clones harbored either 1 (89.8%) or 2 (10.2%) vector copies per cell and displayed partial to full correction of ADA expression, purine metabolism, and T-cell receptor-driven functions. Analysis of RV integration sites indicated a high diversity in T-cell origin, consistently with the polyclonal T-cell receptor-Vbeta repertoire. Quantitative transcript analysis of 120 genes within a 200-kb window around RV integration sites showed modest (2.8- to 5.2-fold) dysregulation of 5.8% genes in 18.6% of the T-cell clones compared with controls. Nonetheless, affected clones maintained a stable phenotype and normal in vitro functions. These results confirm that RV-mediated gene transfer for ADA-SCID is safe, and provide crucial information for the development of future gene therapy protocols. The trials described herein have been registered at http://www.clinicaltrials.gov as #NCT00598481 and #NCT00599781.

Integration of retroviral vectors induces minor changes in the transcriptional activity of T cells from ADA-SCID patients treated with gene therapy / B., Cassani; E., Montini; Maruggi, Giulietta; A., Ambrosi; M., Mirolo; S., Selleri; E., Biral; I., Frugnoli; V., Hernandez Trujillo; C., Di Serio; M. G., Roncarolo; L., Naldini; Mavilio, Fulvio; A., Aiuti. - In: BLOOD. - ISSN 0006-4971. - STAMPA. - 114:17(2009), pp. 3546-3556. [10.1182/blood-2009-02-202085]

Integration of retroviral vectors induces minor changes in the transcriptional activity of T cells from ADA-SCID patients treated with gene therapy

MARUGGI, Giulietta;MAVILIO, Fulvio;
2009

Abstract

Gene transfer into hematopoietic stem cells by gamma-retroviral vectors (RVs) is an effective treatment for inherited blood disorders, although potentially limited by the risk of insertional mutagenesis. We evaluated the genomic impact of RV integration in T lymphocytes from adenosine deaminase-deficient severe combined immunodeficiency (ADA-SCID) patients 10 to 30 months after infusion of autologous, genetically corrected CD34(+) cells. Expression profiling on ex vivo T-cell bulk population revealed no difference with respect to healthy controls. To assess the effect of vector integration on gene expression at the single-cell level, primary T-cell clones were isolated from 2 patients. T-cell clones harbored either 1 (89.8%) or 2 (10.2%) vector copies per cell and displayed partial to full correction of ADA expression, purine metabolism, and T-cell receptor-driven functions. Analysis of RV integration sites indicated a high diversity in T-cell origin, consistently with the polyclonal T-cell receptor-Vbeta repertoire. Quantitative transcript analysis of 120 genes within a 200-kb window around RV integration sites showed modest (2.8- to 5.2-fold) dysregulation of 5.8% genes in 18.6% of the T-cell clones compared with controls. Nonetheless, affected clones maintained a stable phenotype and normal in vitro functions. These results confirm that RV-mediated gene transfer for ADA-SCID is safe, and provide crucial information for the development of future gene therapy protocols. The trials described herein have been registered at http://www.clinicaltrials.gov as #NCT00598481 and #NCT00599781.
2009
114
17
3546
3556
Integration of retroviral vectors induces minor changes in the transcriptional activity of T cells from ADA-SCID patients treated with gene therapy / B., Cassani; E., Montini; Maruggi, Giulietta; A., Ambrosi; M., Mirolo; S., Selleri; E., Biral; I., Frugnoli; V., Hernandez Trujillo; C., Di Serio; M. G., Roncarolo; L., Naldini; Mavilio, Fulvio; A., Aiuti. - In: BLOOD. - ISSN 0006-4971. - STAMPA. - 114:17(2009), pp. 3546-3556. [10.1182/blood-2009-02-202085]
B., Cassani; E., Montini; Maruggi, Giulietta; A., Ambrosi; M., Mirolo; S., Selleri; E., Biral; I., Frugnoli; V., Hernandez Trujillo; C., Di Serio; M. G., Roncarolo; L., Naldini; Mavilio, Fulvio; A., Aiuti
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/639569
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