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An experimental system for the evaluation of retroviral vector design to diminish the risk for proto-oncogene activation.
Blood. 2008 Feb 15; 111(4):1866-75.Blood

Abstract

Pathogenic activation of the LMO2 proto-oncogene by an oncoretroviral vector insertion in a clinical trial for X-linked severe combined immunodeficiency (X-SCID) has prompted safety concerns. We used an adeno-associated virus vector to achieve targeted insertion of a gamma-retroviral long terminal repeat (LTR) driving a GFP expression cassette with flanking loxP sites in a human T-cell line at the precise location of vector integration in one of the patients with X-SCID. The LTR-GFP cassette was inserted into the first intron of the LMO2 gene, resulting in strong activation of LMO2. Cre-mediated cassette exchange was used to replace the original LTR-GFP cassette with one flanked by insulator elements leading to a several fold reduction in LMO2 expression. The LTR-GFP cassette was also replaced with a globin gene regulatory cassette that failed to activate the LMO2 gene in lymphoid cells. A gamma-retroviral vector with 2 intact LTRs resulted in activation of the LMO2 gene when inserted into the first intron, but a self-inactivating lentiviral vector with an internal cellular promoter and flanking insulator elements did not activate the LMO2 gene. Thus, this system is useful for comparing the safety profiles of vector cassettes with various regulatory elements for their potential for proto-oncogene activation.

Authors+Show Affiliations

Department of Hematology, St Jude Children's Research Hospital, Memphis, TN 38105, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17991809

Citation

Ryu, Byoung Y., et al. "An Experimental System for the Evaluation of Retroviral Vector Design to Diminish the Risk for Proto-oncogene Activation." Blood, vol. 111, no. 4, 2008, pp. 1866-75.
Ryu BY, Evans-Galea MV, Gray JT, et al. An experimental system for the evaluation of retroviral vector design to diminish the risk for proto-oncogene activation. Blood. 2008;111(4):1866-75.
Ryu, B. Y., Evans-Galea, M. V., Gray, J. T., Bodine, D. M., Persons, D. A., & Nienhuis, A. W. (2008). An experimental system for the evaluation of retroviral vector design to diminish the risk for proto-oncogene activation. Blood, 111(4), 1866-75.
Ryu BY, et al. An Experimental System for the Evaluation of Retroviral Vector Design to Diminish the Risk for Proto-oncogene Activation. Blood. 2008 Feb 15;111(4):1866-75. PubMed PMID: 17991809.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An experimental system for the evaluation of retroviral vector design to diminish the risk for proto-oncogene activation. AU - Ryu,Byoung Y, AU - Evans-Galea,Marguerite V, AU - Gray,John T, AU - Bodine,David M, AU - Persons,Derek A, AU - Nienhuis,Arthur W, Y1 - 2007/11/08/ PY - 2007/11/10/pubmed PY - 2008/5/22/medline PY - 2007/11/10/entrez SP - 1866 EP - 75 JF - Blood JO - Blood VL - 111 IS - 4 N2 - Pathogenic activation of the LMO2 proto-oncogene by an oncoretroviral vector insertion in a clinical trial for X-linked severe combined immunodeficiency (X-SCID) has prompted safety concerns. We used an adeno-associated virus vector to achieve targeted insertion of a gamma-retroviral long terminal repeat (LTR) driving a GFP expression cassette with flanking loxP sites in a human T-cell line at the precise location of vector integration in one of the patients with X-SCID. The LTR-GFP cassette was inserted into the first intron of the LMO2 gene, resulting in strong activation of LMO2. Cre-mediated cassette exchange was used to replace the original LTR-GFP cassette with one flanked by insulator elements leading to a several fold reduction in LMO2 expression. The LTR-GFP cassette was also replaced with a globin gene regulatory cassette that failed to activate the LMO2 gene in lymphoid cells. A gamma-retroviral vector with 2 intact LTRs resulted in activation of the LMO2 gene when inserted into the first intron, but a self-inactivating lentiviral vector with an internal cellular promoter and flanking insulator elements did not activate the LMO2 gene. Thus, this system is useful for comparing the safety profiles of vector cassettes with various regulatory elements for their potential for proto-oncogene activation. SN - 0006-4971 UR - https://www.unboundmedicine.com/medline/citation/17991809/An_experimental_system_for_the_evaluation_of_retroviral_vector_design_to_diminish_the_risk_for_proto_oncogene_activation_ L2 - https://ashpublications.org/blood/article-lookup/doi/10.1182/blood-2007-04-085506 DB - PRIME DP - Unbound Medicine ER -