@misc{10481/84712,
year = {2023},
month = {6},
url = {https://hdl.handle.net/10481/84712},
abstract = {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.},
organization = {Spanish ISCIII Health Research Fund},
organization = {European Union (EU)	PI18/00337
PI21/00298
RD21/0017/0004
PI18/00330
PI17/00672},
organization = {Red TerAv},
organization = {Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia},
organization = {Spanish Government	PI18/00337
PI21/00298},
organization = {European Union-NextGenerationEU - Maria Zambrano Senior Program	RD21/0017/0004
PI18/00330
PI17/00672},
organization = {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},
organization = {European Cooperation in Science and Technology (COST)	00123009/SNEO-20191072},
organization = {MINECO - European Regional Development Fund	PLEC2021-008094},
organization = {Spanish Government	0006/2018},
organization = {FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades	CA21113},
organization = {Spanish Government	SAF2015-71589-P},
organization = {MCI	RYC-2016-21395},
organization = {German Research Foundation (DFG)	PY20_00619 y A-CTS-28_UGR20},
organization = {Biomedicine Program of the University of Granada (Spain)	FPU16/05467
FPU17/02268
FPU17/04327
DIN2018-010180
DIN2020-011550
PEJ-2018-001760-A},
publisher = {CellPress},
title = {Lentiviral vectors for inducible, transactivator-free advanced therapy medicinal products: Application to CAR-T cells},
doi = {10.1016/j.omtn.2023.03.018.},
author = {Tristán Manzano, María and Maldonado Pérez, Noelia and Justicia Lirio, Pedro and Cortijo Gutiérrez, Marina and Tristán Ramos, Pablo and Blanco Benítez, Carlos and Pavlovic, Kristina and Aguilar González, Araceli and Muñoz Fernández, Pilar and Molina Estévez, Francisco Javier and Griesche, Valerie and Marchal Corrales, Juan Antonio and Heras, Sara R. and Benabdellah, Karim and Martín Molina, Francisco},
}