Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance
A naturally occurring 32-base pair deletion of the HIV-1 co-receptor CCR5 has demonstrated protection against HIV infection of human CD4+ T cells. Recent genetic engineering approaches using engineered nucleases to disrupt the gene and mimic this mutation show promise for HIV therapy. We developed a...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2016-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253117300720 |
| _version_ | 1828793106466603008 |
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| author | Guillermo S Romano Ibarra Biswajit Paul Blythe D Sather Patrick M Younan Karen Sommer John P Kowalski Malika Hale Barry Stoddard Jordan Jarjour Alexander Astrakhan Hans-Peter Kiem David J Rawlings |
| author_facet | Guillermo S Romano Ibarra Biswajit Paul Blythe D Sather Patrick M Younan Karen Sommer John P Kowalski Malika Hale Barry Stoddard Jordan Jarjour Alexander Astrakhan Hans-Peter Kiem David J Rawlings |
| author_sort | Guillermo S Romano Ibarra |
| collection | DOAJ |
| description | A naturally occurring 32-base pair deletion of the HIV-1 co-receptor CCR5 has demonstrated protection against HIV infection of human CD4+ T cells. Recent genetic engineering approaches using engineered nucleases to disrupt the gene and mimic this mutation show promise for HIV therapy. We developed a megaTAL nuclease targeting the third extracellular loop of CCR5 that we delivered to primary human T cells by mRNA transfection. The CCR5 megaTAL nuclease established resistance to HIV in cell lines and disrupted the expression of CCR5 on primary human CD4+ T cells with a high efficiency, achieving up to 80% modification of the locus in primary cells as measured by molecular analysis. Gene-modified cells engrafted at levels equivalent to unmodified cells when transplanted into immunodeficient mice. Furthermore, genetically modified CD4+ cells were preferentially expanded during HIV-1 infection in vivo in an immunodeficient mouse model. Our results demonstrate the feasibility of targeting CCR5 in primary T cells using an engineered megaTAL nuclease, and the potential to use gene-modified cells to reconstitute a patient's immune system and provide protection from HIV infection. |
| first_indexed | 2024-12-12T03:21:39Z |
| format | Article |
| id | doaj.art-9a753dd3c9794ca9b239d6c43c9c7b98 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-12T03:21:39Z |
| publishDate | 2016-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-9a753dd3c9794ca9b239d6c43c9c7b982022-12-22T00:40:09ZengElsevierMolecular Therapy: Nucleic Acids2162-25312016-01-015C10.1038/mtna.2016.56Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 ResistanceGuillermo S Romano Ibarra0Biswajit Paul1Blythe D Sather2Patrick M Younan3Karen Sommer4John P Kowalski5Malika Hale6Barry Stoddard7Jordan Jarjour8Alexander Astrakhan9Hans-Peter Kiem10David J Rawlings11Center for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, Washington, USAClinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USACenter for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, Washington, USAClinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USACenter for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, Washington, USAClinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USACenter for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, Washington, USAClinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USAbluebird bio, Inc., Seattle, Washington, USAbluebird bio, Inc., Seattle, Washington, USAClinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USACenter for Immunity and Immunotherapies and Program for Cell and Gene Therapy, Seattle Children's Research Institute, Seattle, Washington, USAA naturally occurring 32-base pair deletion of the HIV-1 co-receptor CCR5 has demonstrated protection against HIV infection of human CD4+ T cells. Recent genetic engineering approaches using engineered nucleases to disrupt the gene and mimic this mutation show promise for HIV therapy. We developed a megaTAL nuclease targeting the third extracellular loop of CCR5 that we delivered to primary human T cells by mRNA transfection. The CCR5 megaTAL nuclease established resistance to HIV in cell lines and disrupted the expression of CCR5 on primary human CD4+ T cells with a high efficiency, achieving up to 80% modification of the locus in primary cells as measured by molecular analysis. Gene-modified cells engrafted at levels equivalent to unmodified cells when transplanted into immunodeficient mice. Furthermore, genetically modified CD4+ cells were preferentially expanded during HIV-1 infection in vivo in an immunodeficient mouse model. Our results demonstrate the feasibility of targeting CCR5 in primary T cells using an engineered megaTAL nuclease, and the potential to use gene-modified cells to reconstitute a patient's immune system and provide protection from HIV infection.http://www.sciencedirect.com/science/article/pii/S2162253117300720Gene therapygene editingHIVmegaTALCCR5 |
| spellingShingle | Guillermo S Romano Ibarra Biswajit Paul Blythe D Sather Patrick M Younan Karen Sommer John P Kowalski Malika Hale Barry Stoddard Jordan Jarjour Alexander Astrakhan Hans-Peter Kiem David J Rawlings Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance Molecular Therapy: Nucleic Acids Gene therapy gene editing HIV megaTAL CCR5 |
| title | Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance |
| title_full | Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance |
| title_fullStr | Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance |
| title_full_unstemmed | Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance |
| title_short | Efficient Modification of the CCR5 Locus in Primary Human T Cells With megaTAL Nuclease Establishes HIV-1 Resistance |
| title_sort | efficient modification of the ccr5 locus in primary human t cells with megatal nuclease establishes hiv 1 resistance |
| topic | Gene therapy gene editing HIV megaTAL CCR5 |
| url | http://www.sciencedirect.com/science/article/pii/S2162253117300720 |
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