RT Journal Article
T1 Physiological lentiviral vectors for the generation of improved CAR-T cells.
A1 Tristan-Manzano, Maria
A1 Maldonado-Perez, Noelia
A1 Justicia-Lirio, Pedro
A1 Muñoz, Pilar
A1 Cortijo-Gutierrez, Marina
A1 Pavlovic, Kristina
A1 Jimenez-Moreno, Rosario
A1 Nogueras, Sonia
A1 Carmona, M Dolores
A1 Sanchez-Hernandez, Sabina
A1 Aguilar-Gonzalez, Araceli
A1 Castella, Maria
A1 Juan, Manel
A1 Marañon, Concepcion
A1 Marchal, Juan Antonio
A1 Benabdellah, Karim
A1 Herrera, Concha
A1 Martin, Francisco
K1 CAR-T
K1 T cells phenotype
K1 TCR-like expression
K1 WAS gene promoter
K1 Exhaustion
AB Anti-CD19 chimeric antigen receptor (CAR)-T cells have achieved impressive outcomes for the treatment of relapsed and refractory B-lineage neoplasms. However, important limitations still remain due to severe adverse events (i.e., cytokine release syndrome and neuroinflammation) and relapse of 40%-50% of the treated patients. Most CAR-T cells are generated using retroviral vectors with strong promoters that lead to high CAR expression levels, tonic signaling, premature exhaustion, and overstimulation, reducing efficacy and increasing side effects. Here, we show that lentiviral vectors (LVs) expressing the transgene through a WAS gene promoter (AW-LVs) closely mimic the T cell receptor (TCR)/CD3 expression kinetic upon stimulation. These AW-LVs can generate improved CAR-T cells as a consequence of their moderate and TCR-like expression profile. Compared with CAR-T cells generated with human elongation factor α (EF1α)-driven-LVs, AW-CAR-T cells exhibited lower tonic signaling, higher proportion of naive and stem cell memory T cells, less exhausted phenotype, and milder secretion of tumor necrosis factor alpha (TNF-α) and interferon (IFN)-ɣ after efficient destruction of CD19+ lymphoma cells, both in vitro and in vivo. Moreover, we also showed their improved efficiency using an in vitro CD19+ pancreatic tumor model. We finally demonstrated the feasibility of large-scale manufacturing of AW-CAR-T cells in guanosine monophosphate (GMP)-like conditions. Based on these data, we propose the use of AW-LVs for the generation of improved CAR-T products.
PB Cell Press
SN 2372-7705
YR 2022
FD 2022-05-07
LK http://hdl.handle.net/10668/22464
UL http://hdl.handle.net/10668/22464
LA en
NO Tristán-Manzano M, Maldonado-Pérez N, Justicia-Lirio P, Muñoz P, Cortijo-Gutiérrez M, Pavlovic K, et al.  Physiological lentiviral vectors for the generation of improved CAR-T cells. Mol Ther Oncolytics. 2022 May 18;25:335-349. doi: 10.1016/j.omto.2022.05.003. Erratum in: Mol Ther Oncolytics. 2022 Jul 21;26:245
NO This study was funded by the Spanish ISCIII Health Research Fund and the European Regional Development Fund (FEDER) through research grantsPI15/02015, PI18/00337, PI21/00298, and Red TerAv RD21/0017/0004 (F.M.); PI18/00330 (K.B.); and PI17/00672 (P.M.). The CECEyU and CSyF of the Junta de Andalucía FEDER/European Cohesion Fund (FSE) for Andalusia provided the following research grants: 2016000073391-TRA, 2016000073332-TRA, PI-57069, PA IDI-Bio326, CARTPI-0001-201, PECART-0031-2020, Red RANTECAR CAR-T 2019 00400200101918, and PLEC2021-008094 (F.M.); PI-0014-2016 (K.B.); PEER-0286-2019 (P.M.); and the Ministerio de Ciencia, Innovación y Universidades through research grants 00123009/SNEO-20191072 and PLEC2021-008094 to F.M. K.B. andC.M. held Nicolas Monardes contracts from regional Ministry of Health (no. 0006/2018 and C2-0002-2019, respectively). M.T.-M., N.M.-P., and A.A.-G. are funded by Spanish Ministry of Education and Science through fellowships FPU16/05467, FPU17/02268, and FPU17/04327, respectively. P.-J.L. is funded through an industrial doctorate MCI DIN2018-010180 to LentiStem Biotech. P.M. is funded by Fundación Andaluza Progreso y Salud. M.C.-G. is funded by Spanish Ministry of Education and Science through fellowship GJ(PEJ-2018-001760-A). M.D.C. is funded by the grant PE-0223-2018from CSyF of the Junta de Andalucia. M.T.-M., N.M.-P., P.-J.L., M.C.-G., and A.A.-G. are PhD students from the Biomedicine Programme of the University of Granada (Spain).
DS RISalud
RD Apr 5, 2025