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Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors

Adverse events linked to perturbations of cellular genes by vector insertion reported in gene therapy trials and animal models have prompted attempts to better understand the mechanisms directing viral vector integration. The integration profiles of vectors based on MLV, ASLV, SIV, and HIV have all...

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Autores principales: Métais, Jean-Yves, Topp, Shana, Doty, Raymond T., Borate, Bhavesh, Nguyen, Anh-Dao, Wolfsberg, Tyra G., Abkowitz, Janis L., Dunbar, Cynthia E.
Formato: Texto
Lenguaje:English
Publicado: 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2883645/
https://www.ncbi.nlm.nih.gov/pubmed/20237508
http://dx.doi.org/10.1038/gt.2010.24
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author Métais, Jean-Yves
Topp, Shana
Doty, Raymond T.
Borate, Bhavesh
Nguyen, Anh-Dao
Wolfsberg, Tyra G.
Abkowitz, Janis L.
Dunbar, Cynthia E.
author_facet Métais, Jean-Yves
Topp, Shana
Doty, Raymond T.
Borate, Bhavesh
Nguyen, Anh-Dao
Wolfsberg, Tyra G.
Abkowitz, Janis L.
Dunbar, Cynthia E.
author_sort Métais, Jean-Yves
collection PubMed
description Adverse events linked to perturbations of cellular genes by vector insertion reported in gene therapy trials and animal models have prompted attempts to better understand the mechanisms directing viral vector integration. The integration profiles of vectors based on MLV, ASLV, SIV, and HIV have all been shown to be non-random, and novel vectors with a safer integration pattern have been sought. Recently we developed a producer cell line called CatPac that packages standard MoMLV vectors with FeLV gag, pol and env gene products. We now report the integration profile of this vector, asking if the FeLV integrase and capsid proteins could modify the MoMLV integration profile, potentially resulting in a less genotoxic pattern. We transduced rhesus macaque CD34+ hematopoietic progenitor cells with CatPac or standard MoMLV vectors, and determined their integration profile by LAM-PCR. We obtained 184 and 175 unique integration sites (IS) respectively for CatPac and standard MoMLV vectors, and these were compared to 10 000 in silico-generated random IS. The integration profile for CatPac vector was similar to MoMLV and equally non-random, with a propensity for integration near transcription start sites and in highly dense gene regions. We found an IS for CatPac vector localized 715 nucleotides upstream of LMO-2, the gene involved in the ALL developed by X-SCID patients treated via gene therapy using MoMLV vectors. In conclusion, we found that replacement of MoMLV env, gag, and pol gene products with FeLV did not alter the basic integration profile. Thus there appears to be no safety advantage for this packaging system. However, considering the stability and efficacy of CatPac vectors, further development is warranted, utilizing potentially safer vector backbones, for instance those with a SIN configuration.
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spelling pubmed-28836452010-12-01 Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors Métais, Jean-Yves Topp, Shana Doty, Raymond T. Borate, Bhavesh Nguyen, Anh-Dao Wolfsberg, Tyra G. Abkowitz, Janis L. Dunbar, Cynthia E. Gene Ther Article Adverse events linked to perturbations of cellular genes by vector insertion reported in gene therapy trials and animal models have prompted attempts to better understand the mechanisms directing viral vector integration. The integration profiles of vectors based on MLV, ASLV, SIV, and HIV have all been shown to be non-random, and novel vectors with a safer integration pattern have been sought. Recently we developed a producer cell line called CatPac that packages standard MoMLV vectors with FeLV gag, pol and env gene products. We now report the integration profile of this vector, asking if the FeLV integrase and capsid proteins could modify the MoMLV integration profile, potentially resulting in a less genotoxic pattern. We transduced rhesus macaque CD34+ hematopoietic progenitor cells with CatPac or standard MoMLV vectors, and determined their integration profile by LAM-PCR. We obtained 184 and 175 unique integration sites (IS) respectively for CatPac and standard MoMLV vectors, and these were compared to 10 000 in silico-generated random IS. The integration profile for CatPac vector was similar to MoMLV and equally non-random, with a propensity for integration near transcription start sites and in highly dense gene regions. We found an IS for CatPac vector localized 715 nucleotides upstream of LMO-2, the gene involved in the ALL developed by X-SCID patients treated via gene therapy using MoMLV vectors. In conclusion, we found that replacement of MoMLV env, gag, and pol gene products with FeLV did not alter the basic integration profile. Thus there appears to be no safety advantage for this packaging system. However, considering the stability and efficacy of CatPac vectors, further development is warranted, utilizing potentially safer vector backbones, for instance those with a SIN configuration. 2010-03-18 2010-06 /pmc/articles/PMC2883645/ /pubmed/20237508 http://dx.doi.org/10.1038/gt.2010.24 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Métais, Jean-Yves
Topp, Shana
Doty, Raymond T.
Borate, Bhavesh
Nguyen, Anh-Dao
Wolfsberg, Tyra G.
Abkowitz, Janis L.
Dunbar, Cynthia E.
Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors
title Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors
title_full Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors
title_fullStr Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors
title_full_unstemmed Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors
title_short Feline Leukemia Virus Integrase And Capsid Packaging Functions Do Not Change The Insertion Profile Of Standard Moloney Retroviral Vectors
title_sort feline leukemia virus integrase and capsid packaging functions do not change the insertion profile of standard moloney retroviral vectors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2883645/
https://www.ncbi.nlm.nih.gov/pubmed/20237508
http://dx.doi.org/10.1038/gt.2010.24
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