Discovery and preclinical development of a SdAb-based CAR-T technology for targeting CD33 in AML

Chimeric antigen receptor T cell (CAR-T) therapies have revolutionized cancer immunotherapy. Traditional single-chain variable fragments (ScFvs) used as CAR recognition moieties face challenges such as high tonic signaling, compromised binding epitopes, and suboptimal affinity. Single-domain antibod...

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Detalles Bibliográficos
Autores: Bernasconi-Bisio, Franco, Molina, Eva, Ibarra, Vianca, Ibáñez-Sala, Inés, Rochira, Federica, Jauregui, Patricia, Rodríguez-Diaz, Saray, Martínez-Turrillas, Rebeca, Azagra-Barber, Iñigo, Gómez-Cebrián, Nuria, Lasarte, Juan José, Puchades Carrasco, Leonor, Vanrell, Lucía, Rodríguez-Madoz, Juan Roberto, Prosper, Felipe, Pineda Lucena, Antonio
Tipo de recurso: artículo
Fecha de publicación:2025
País:España
Institución:Universidad Católica de Valencia San Vicente Mártir
Repositorio:RIUCV. Repositorio de la Universidad Católica de Valencia San Vicente Mártir
Idioma:inglés
OAI Identifier:oai:riucv.ucv.es:20.500.12466/5480
Acceso en línea:http://hdl.handle.net/20.500.12466/5480
Access Level:acceso abierto
Palabra clave:Regular Issues
Single-domain antibodies
SdAbs
Nanobodies
VHH libraries
CAR-T cell therapy
Acute myeloid leukemia
AML
CD33
Descripción
Sumario:Chimeric antigen receptor T cell (CAR-T) therapies have revolutionized cancer immunotherapy. Traditional single-chain variable fragments (ScFvs) used as CAR recognition moieties face challenges such as high tonic signaling, compromised binding epitopes, and suboptimal affinity. Single-domain antibodies (SdAbs) offer an attractive alternative due to their smaller size, stability, and reduced immunogenicity. In this work, we developed an SdAb-CAR-T cell discovery platform integrating generation, characterization, and selection of SdAbs based on various properties. This approach was demonstrated by developing CAR-T cells with SdAbs against CD33, a target for acute myeloid leukemia (AML). We identified diverse SdAbs against CD33, with affinities ranging from 3.9–115 nM, and characterized their binding kinetics and epitope recognition. Using SdAb-based second-generation CARs, we assessed tonic signaling, T cell phenotypes, cytotoxicity and cytokine release in vitro, resulting in reduced tonic signaling and increased cytokine production. In vivo, SdAb-based CAR-T cells exhibited enhanced efficacy at lower doses, in a xenograft AML mouse model, demonstrating advantages over ScFv-based CD33 CAR-T cells.

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