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Lentiviral vector integration sites in human NOD/SCID repopulating cells.
J Gene Med. 2006 Oct; 8(10):1197-207.JG

Abstract

BACKGROUND

Recent observations of insertional mutagenesis in preclinical and clinical settings emphasize the relevance of investigating comprehensively the spectrum of integration sites targeted by specific vectors.

METHODS

We followed the engraftment of lentivirally transduced human cord blood (CB) progenitor cells after transplantation into NOD/SCID mice using a self-inactivating HIV-1-derived vector expressing the enhanced green fluorescent protein (EGFP).

RESULTS

The mean of transduction of CD34(+) CB cells was 41%, as deduced from the percentage of EGFP(+) cells before transplantation. At 3 weeks post-transplantation, the average of EGFP(+) cells in the human cell population was 65 +/- 8%, and increased to 75 +/- 10% at 12 weeks post-transplantation. In order to determine the proviral integration sites in human NOD/SCID repopulating cells (SRCs) we used the ligation-mediated polymerase chain reaction (LM-PCR) technique. Sixty-eight percent of the integrations were found to be located in RefSeq genes, most of them in intron regions. Twenty percent of these integrations occurred within a distance of 10 kb from the transcription start site; a percentage that is significantly lower compared to that observed in cells transduced by gammaretroviral vectors. Sixty-two percent of integrations occurred in genes with a biological function in cell metabolism, and four integrations were located in genes with a role in tumorigenesis.

CONCLUSIONS

These investigations indicate that integration of lentiviral vectors in human repopulating cells capable of engrafting NOD/SCID mice preferentially occur in coding regions of the human genome. Nevertheless, the clustering of integrations at the transcriptional start is not as high as that observed for gammaretroviral vectors.

Authors+Show Affiliations

Research Program Innovative Cancer Diagnostics and Therapy, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany. s.laufs@dkfz.deNo 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

16960916

Citation

Laufs, Stephanie, et al. "Lentiviral Vector Integration Sites in Human NOD/SCID Repopulating Cells." The Journal of Gene Medicine, vol. 8, no. 10, 2006, pp. 1197-207.
Laufs S, Guenechea G, Gonzalez-Murillo A, et al. Lentiviral vector integration sites in human NOD/SCID repopulating cells. J Gene Med. 2006;8(10):1197-207.
Laufs, S., Guenechea, G., Gonzalez-Murillo, A., Zsuzsanna Nagy, K., Luz Lozano, M., del Val, C., Jonnakuty, S., Hotz-Wagenblatt, A., Jens Zeller, W., Bueren, J. A., & Fruehauf, S. (2006). Lentiviral vector integration sites in human NOD/SCID repopulating cells. The Journal of Gene Medicine, 8(10), 1197-207.
Laufs S, et al. Lentiviral Vector Integration Sites in Human NOD/SCID Repopulating Cells. J Gene Med. 2006;8(10):1197-207. PubMed PMID: 16960916.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Lentiviral vector integration sites in human NOD/SCID repopulating cells. AU - Laufs,Stephanie, AU - Guenechea,Guillermo, AU - Gonzalez-Murillo,Africa, AU - Zsuzsanna Nagy,K, AU - Luz Lozano,M, AU - del Val,Coral, AU - Jonnakuty,Sunitha, AU - Hotz-Wagenblatt,Agnes, AU - Jens Zeller,W, AU - Bueren,Juan A, AU - Fruehauf,Stefan, PY - 2006/9/9/pubmed PY - 2007/2/1/medline PY - 2006/9/9/entrez SP - 1197 EP - 207 JF - The journal of gene medicine JO - J Gene Med VL - 8 IS - 10 N2 - BACKGROUND: Recent observations of insertional mutagenesis in preclinical and clinical settings emphasize the relevance of investigating comprehensively the spectrum of integration sites targeted by specific vectors. METHODS: We followed the engraftment of lentivirally transduced human cord blood (CB) progenitor cells after transplantation into NOD/SCID mice using a self-inactivating HIV-1-derived vector expressing the enhanced green fluorescent protein (EGFP). RESULTS: The mean of transduction of CD34(+) CB cells was 41%, as deduced from the percentage of EGFP(+) cells before transplantation. At 3 weeks post-transplantation, the average of EGFP(+) cells in the human cell population was 65 +/- 8%, and increased to 75 +/- 10% at 12 weeks post-transplantation. In order to determine the proviral integration sites in human NOD/SCID repopulating cells (SRCs) we used the ligation-mediated polymerase chain reaction (LM-PCR) technique. Sixty-eight percent of the integrations were found to be located in RefSeq genes, most of them in intron regions. Twenty percent of these integrations occurred within a distance of 10 kb from the transcription start site; a percentage that is significantly lower compared to that observed in cells transduced by gammaretroviral vectors. Sixty-two percent of integrations occurred in genes with a biological function in cell metabolism, and four integrations were located in genes with a role in tumorigenesis. CONCLUSIONS: These investigations indicate that integration of lentiviral vectors in human repopulating cells capable of engrafting NOD/SCID mice preferentially occur in coding regions of the human genome. Nevertheless, the clustering of integrations at the transcriptional start is not as high as that observed for gammaretroviral vectors. SN - 1099-498X UR - https://www.unboundmedicine.com/medline/citation/16960916/Lentiviral_vector_integration_sites_in_human_NOD/SCID_repopulating_cells_ L2 - https://doi.org/10.1002/jgm.958 DB - PRIME DP - Unbound Medicine ER -