Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells
Controlling transgene expression through an externally administered inductor is envisioned as a potent strategy to improve safety and efficacy of gene therapy approaches. Generally, inducible ON systems require a chimeric transcription factor (transactivator) that becomes activated by an inductor, w...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S216225312300077X |
| _version_ | 1827964240736026624 |
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| author | María Tristán-Manzano Noelia Maldonado-Pérez Pedro Justicia-Lirio Marina Cortijo-Gutierréz Pablo Tristán-Ramos Carlos Blanco-Benítez Kristina Pavlovic Araceli Aguilar-González Pilar Muñoz Francisco J. Molina-Estevez Valerie Griesche Juan Antonio Marchal Sara R. Heras Karim Benabdellah Francisco Martin |
| author_facet | María Tristán-Manzano Noelia Maldonado-Pérez Pedro Justicia-Lirio Marina Cortijo-Gutierréz Pablo Tristán-Ramos Carlos Blanco-Benítez Kristina Pavlovic Araceli Aguilar-González Pilar Muñoz Francisco J. Molina-Estevez Valerie Griesche Juan Antonio Marchal Sara R. Heras Karim Benabdellah Francisco Martin |
| author_sort | María Tristán-Manzano |
| collection | DOAJ |
| description | Controlling transgene expression through an externally administered inductor is envisioned as a potent strategy to improve safety and efficacy of gene therapy approaches. Generally, inducible ON systems require a chimeric transcription factor (transactivator) that becomes activated by an inductor, which is not optimal for clinical translation due to their toxicity. We generated previously the first all-in-one, transactivator-free, doxycycline (Dox)-responsive (Lent-On-Plus or LOP) lentiviral vectors (LVs) able to control transgene expression in human stem cells. Here, we have generated new versions of the LOP LVs and have analyzed their applicability for the generation of inducible advanced therapy medicinal products (ATMPs) with special focus on primary human T cells. We have shown that, contrary to all other cell types analyzed, an Is2 insulator must be inserted into the 3′ long terminal repeat of the LOP LVs in order to control transgene expression in human primary T cells. Importantly, inducible primary T cells generated by the LOPIs2 LVs are responsive to ultralow doses of Dox and have no changes in phenotype or function compared with untransduced T cells. We validated the LOPIs2 system by generating inducible CAR-T cells that selectively kill CD19+ cells in the presence of Dox. In summary, we describe here the first transactivator-free, all-one-one system capable of generating Dox-inducible ATMPs. |
| first_indexed | 2024-04-09T17:13:46Z |
| format | Article |
| id | doaj.art-5e23d31a5c5e4bcd886b94089d46f8e8 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-09T17:13:46Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-5e23d31a5c5e4bcd886b94089d46f8e82023-04-20T04:36:18ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-06-0132322339Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cellsMaría Tristán-Manzano0Noelia Maldonado-Pérez1Pedro Justicia-Lirio2Marina Cortijo-Gutierréz3Pablo Tristán-Ramos4Carlos Blanco-Benítez5Kristina Pavlovic6Araceli Aguilar-González7Pilar Muñoz8Francisco J. Molina-Estevez9Valerie Griesche10Juan Antonio Marchal11Sara R. Heras12Karim Benabdellah13Francisco Martin14LentiStem Biotech, Pfizer-University of Granada-Junta de Andalucía Centre for Genomics and Oncological Research (GENYO), PTS, Avda. de la Ilustración 114, 18016 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Departamento de Bioquimica y Biología Molecular III e Inmunologia, Facultad de Medicina, Universidad de Granada, PTS Granada – Avenida de la Investigación, 11, 18006 Granada, SpainLentiStem Biotech, Pfizer-University of Granada-Junta de Andalucía Centre for Genomics and Oncological Research (GENYO), PTS, Avda. de la Ilustración 114, 18016 Granada, Spain; Department of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Department de Bioquimica y Biología Molecular II, Faculty of Pharmacy, University of Granada, Campus Universitario de Cartuja, 18071 Granada, SpainLentiStem Biotech, Pfizer-University of Granada-Junta de Andalucía Centre for Genomics and Oncological Research (GENYO), PTS, Avda. de la Ilustración 114, 18016 Granada, Spain; Department of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Maimonides Institute of Biomedical Research in Cordoba (IMIBIC), Cellular Therapy Unit, Reina Sofía University Hospital, University of Cordoba, Cordoba, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Department of Medicinal & Organic Chemistry and Excellence Research Unit of “Chemistry Applied to Biomedicine and the Environment”, Faculty of Pharmacy, University of Granada, Campus de Cartuja s/n, 18071 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Department de Biología Celular, Facultad de Ciencias, Universidad de Granada, Campus Fuentenueva, 18071 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; DKFZ. German Cancer Research Center, Heidelberg, Baden-Wurtemberg, GermanyInstituto de Investigación Biosanitaria ibs.GRANADA, University Hospitals of Granada – University of Granada, 18016 Granada, Spain; Excellence Research Unit “Modeling Nature” (MNat), University of Granada, 18016 Granada, Spain; Biopathology and Regenerative Medicine Institute (IBIMER), Center for Biomedicinal Research (CIBM), University of Granada, 18016 Granada, Spain; Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, Biosanitary Research Institute of Granada (ibs.GRANADA), Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Department de Bioquimica y Biología Molecular II, Faculty of Pharmacy, University of Granada, Campus Universitario de Cartuja, 18071 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, SpainDepartment of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain; Departamento de Bioquimica y Biología Molecular III e Inmunologia, Facultad de Medicina, Universidad de Granada, PTS Granada – Avenida de la Investigación, 11, 18006 Granada, Spain; Instituto de Investigación Biosanitaria ibs.GRANADA, University Hospitals of Granada – University of Granada, 18016 Granada, Spain; Excellence Research Unit “Modeling Nature” (MNat), University of Granada, 18016 Granada, Spain; Corresponding author: Francisco Martin, Department of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain.Controlling transgene expression through an externally administered inductor is envisioned as a potent strategy to improve safety and efficacy of gene therapy approaches. Generally, inducible ON systems require a chimeric transcription factor (transactivator) that becomes activated by an inductor, which is not optimal for clinical translation due to their toxicity. We generated previously the first all-in-one, transactivator-free, doxycycline (Dox)-responsive (Lent-On-Plus or LOP) lentiviral vectors (LVs) able to control transgene expression in human stem cells. Here, we have generated new versions of the LOP LVs and have analyzed their applicability for the generation of inducible advanced therapy medicinal products (ATMPs) with special focus on primary human T cells. We have shown that, contrary to all other cell types analyzed, an Is2 insulator must be inserted into the 3′ long terminal repeat of the LOP LVs in order to control transgene expression in human primary T cells. Importantly, inducible primary T cells generated by the LOPIs2 LVs are responsive to ultralow doses of Dox and have no changes in phenotype or function compared with untransduced T cells. We validated the LOPIs2 system by generating inducible CAR-T cells that selectively kill CD19+ cells in the presence of Dox. In summary, we describe here the first transactivator-free, all-one-one system capable of generating Dox-inducible ATMPs.http://www.sciencedirect.com/science/article/pii/S216225312300077XMT: Delivery Strategiesgene therapyadvance therapy medicinal productsexternally controlledinducibledoxycycline |
| spellingShingle | María Tristán-Manzano Noelia Maldonado-Pérez Pedro Justicia-Lirio Marina Cortijo-Gutierréz Pablo Tristán-Ramos Carlos Blanco-Benítez Kristina Pavlovic Araceli Aguilar-González Pilar Muñoz Francisco J. Molina-Estevez Valerie Griesche Juan Antonio Marchal Sara R. Heras Karim Benabdellah Francisco Martin Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells Molecular Therapy: Nucleic Acids MT: Delivery Strategies gene therapy advance therapy medicinal products externally controlled inducible doxycycline |
| title | Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells |
| title_full | Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells |
| title_fullStr | Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells |
| title_full_unstemmed | Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells |
| title_short | Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells |
| title_sort | lentiviral vectors for inducible transactivator free advanced therapy medicinal products application to car t cells |
| topic | MT: Delivery Strategies gene therapy advance therapy medicinal products externally controlled inducible doxycycline |
| url | http://www.sciencedirect.com/science/article/pii/S216225312300077X |
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