Unveiling the influence of CAR-negative T-cells: enhancing efficacy and ensuring safety in CAR-T therapies
Year: 2025
Type: article
Abstract: BACKGROUND: Despite the promising results of CAR-T cell therapy, still a significant proportion of patients experience treatment failure due to absence of response, progression or treatment-related toxicities. Our research aims to investigate how the CAR-T cell product characteristics and particularly the percentage of CAR-negative cells present in the infusion product affects its efficacy and safety. METHODS: CAR-positive (CAR-pos) and CAR-negative (CAR-neg) cells were analyzed during in vitro expansion with IL-2 or IL-7/IL-15. FACS-sorted populations were assessed for cytotoxic effect and secretion of cytokines. Transcriptomic analyses were conducted for CAR-pos and CAR-neg populations. RESULTS: A significant reduction in the percentage of transduced (CAR-pos) T cells was observed throughout the in vitro cell culture, where CAR-neg T cells displayed greater expansion along the procedure compared to CAR-pos T cells, using either IL-2 or IL-7/IL-15. CAR-pos T cells exhibited increased expression of activation and exhaustion markers. Comparative analyses of CAR-pos and CAR-neg showed an increased cytotoxic effect in purified samples with 100% CAR-pos T cells compared to non-purified product. Quantification of cytokine levels revealed a significant decrease in IL-6 in purified compared to non-purified products. RNA sequencing analysis revealed significant differential gene expression profile between CAR-pos and CAR-neg cells, the prior exhibiting an upregulation of genes associated with immune checkpoint regulation, cytokine-cytokine receptor interaction and Th1/Th2 and Th17 cell differentiation pathways. CONCLUSION: CAR-neg T cells exhibit a significantly greater expansion compared to CAR-pos T cells alongside CAR-T cell production. The percentage of CAR-neg cells influences the specific cytotoxicity of the product and on the cytokine pattern, which might influence both efficacy and toxicity and, therefore, the number of CAR-neg T cells within the infusion product must be considered in the clinical setting. Purification of CAR-pos T cells could potentially optimize product characteristics.
Authors: Belén Sierro-Martínez, Beatriz Guijarro-Albaladejo, Ricardo Fernández-Cisnal, Alfonso Rodríguez-Gil, Paola Hernández-Díaz, María de la Rosa-Garrido, Inmaculada Carrasco-Brocal, Maribel Lara-Chica, Virginia Escamilla-Gómez, Raquel Muñoz-García, Ángela Bella, Francisco Jose García-Cózar, Jose Antonio Pérez-Simón, Estefanía García-Guerrero
Language: English
Topic: CAR-T cell therapy research
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $3,890
Awards: Mejora de la eficiencia de las células CAR-T mediante la optimización de la identificación de antígenos y el aumento de la persistencia de células CAR-T a través de la expresión de señales anti-apoptóticas, G2458631628, EFECTO DE DERIVADOS CANNABIONIDES EN EL METABOLISMO LINFOCITARIO: PAPEL EN EL TRASPLANTE ALOGENICO y LA TERAPIA CON LINFOCITOS T CAR, G3024517585, G3704284177, Células CAR-T anti-BCMA para el tratamiento de pacientes con mieloma múltiple en recaída tras trasplante alogénico: expresión de EGFR como mecanismo de monitorizacion y control de las complicaciones derivadas de la terapia CAR-T, G8248449418
OpenAlex ID: W4413311583