@misc{10481/78149,
year = {2022},
month = {10},
url = {https://hdl.handle.net/10481/78149},
abstract = {Autologous T cells expressing the Chimeric Antigen Receptor (CAR) have been
approved as advanced therapy medicinal products (ATMPs) against several
hematological malignancies. However, the generation of patient-specific CART
products delays treatment and precludes standardization. Allogeneic off-theshelf
CAR-T cells are an alternative to simplify this complex and timeconsuming
process. Here we investigated safety and efficacy of knocking out
the TCR molecule in ARI-0001 CAR-T cells, a second generation aCD19 CAR
approved by the Spanish Agency of Medicines and Medical Devices (AEMPS)
under the Hospital Exemption for treatment of patients older than 25 years with
Relapsed/Refractory acute B cell lymphoblastic leukemia (B-ALL). We first
analyzed the efficacy and safety issues that arise during disruption of the TCR
gene using CRISPR/Cas9. We have shown that edition of TRAC locus in T cells
using CRISPR as ribonuleorproteins allows a highly efficient TCR disruption
(over 80%) without significant alterations on T cells phenotype and with an
increased percentage of energetic mitochondria. However, we also found that
efficient TCRKO can lead to on-target large and medium size deletions,
indicating a potential safety risk of this procedure that needs monitoring.
Importantly, TCR edition of ARI-0001 efficiently prevented allogeneic
responses and did not detectably alter their phenotype, while maintaining a
similar anti-tumor activity ex vivo and in vivo compared to unedited ARI-0001 CAR-T cells. In summary, we showed here that, although there are still some
risks of genotoxicity due to genome editing, disruption of the TCR is a feasible
strategy for the generation of functional allogeneic ARI-0001 CAR-T cells. We
propose to further validate this protocol for the treatment of patients that do
not fit the requirements for standard autologous CAR-T cells administration.},
organization = {Spanish ISCIII Health Research Fund},
organization = {European Commission	PI15/02015	
PI18/00337	
PI21/00298},
organization = {Red TerAv RD21/ 0017/0004	
PI18/ 00330	
PI17/00672},
organization = {CECEyU and CSyF of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia	2016000073391-TRA	
2016000073332-TRA	
PI-57069	
PAIDIBio326	
CARTPI-0001- 201	
PECART-0031-2020	
PI0014-2016	
PEER-0286-2019},
organization = {Spanish Government	00123009/SNEO-20191072	
PLEC2021-008094},
organization = {regional Ministry of Health	0006/2018	
C2-0002-2019},
organization = {Spanish Government	FPU16/05467	
FPU17/02268	
FPU17/04327},
organization = {Junta de Andalucia	PECART-00312020},
organization = {Consejeria de Salud y Familias	PECART-0027-2020	
MCI DIN2018-010180	
DIN2020-011550},
publisher = {Frontiers},
keywords = {CAR-T cells},
keywords = {Lymphoma},
keywords = {TCRKO},
keywords = {CRISPR/Cas9},
keywords = {Off-the-shelf},
keywords = {Safety},
keywords = {Large deletions},
title = {Efficacy and safety of universal (TCRKO) ARI-0001 CAR-T cells for the treatment of B-cell lymphoma},
doi = {10.3389/fimmu.2022.1011858},
author = {Maldonado Pérez, Noelia and Tristán Manzano, María and Justicia Lirio, Pedro and Martínez Planes, Elena and Muñoz, Pilar and Pavlovic, Kristina and Cortijo Gutiérrez, Marina and Blanco Benítez, Carlos and Molina Estévez, Francisco Javier and Marañón, Concepción and Benabdellah, Karim and Martín Molina, Francisco},
}