Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells
Metadata
Show full item recordAuthor
Tristán Manzano, María; Maldonado Pérez, Noelia; Justicia Lirio, Pedro; Cortijo Gutiérrez, Marina; Tristán Ramos, Pablo; Blanco Benítez, Carlos; Pavlovic, Kristina; Aguilar González, Araceli; Muñoz Fernández, Pilar; Molina Estévez, Francisco Javier; Griesche, Valerie; Marchal Corrales, Juan Antonio; Heras, Sara R.; Benabdellah, Karim; Martín Molina, FranciscoEditorial
CellPress
Date
2023-06Referencia bibliográfica
Tristán-Manzano, M., Maldonado-Pérez, N., Justicia-Lirio, P., Cortijo-Gutierréz, M., Tristán-Ramos, P., Blanco-Benítez, C., ... & Martin, F. (2023). Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells. Molecular Therapy-Nucleic Acids, 32, 322-339.[https://doi.org/10.1016/j.omtn.2023.03.018.]
Sponsorship
Spanish ISCIII Health Research Fund; European Union (EU) PI18/00337 PI21/00298 RD21/0017/0004 PI18/00330 PI17/00672; Red TerAv; Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia; Spanish Government PI18/00337 PI21/00298; European Union-NextGenerationEU - Maria Zambrano Senior Program RD21/0017/0004 PI18/00330 PI17/00672; Ministry of Health 2016000073332-TRA PI-57069 CARTPI-0001-201 PE-CART-0031-2020 PI-0014-2016 PECART-0027-2020 ProyExcel_00875 PEER-0286-2019; European Cooperation in Science and Technology (COST) 00123009/SNEO-20191072; MINECO - European Regional Development Fund PLEC2021-008094; Spanish Government 0006/2018; FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades CA21113; Spanish Government SAF2015-71589-P; MCI RYC-2016-21395; German Research Foundation (DFG) PY20_00619 y A-CTS-28_UGR20; Biomedicine Program of the University of Granada (Spain) FPU16/05467 FPU17/02268 FPU17/04327 DIN2018-010180 DIN2020-011550 PEJ-2018-001760-AAbstract
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 30 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 transactivatorfree,
all-one-one system capable of generating Dox-inducible
ATMPs.