Data from Modifications to the Framework Regions Eliminate Chimeric Antigen Receptor Tonic Signaling
Year: 2023
Type: other
Abstract: Chimeric antigen receptor (CAR) tonic signaling, defined as spontaneous activation and release of proinflammatory cytokines by CAR-T cells, is considered a negative attribute because it leads to impaired antitumor effects. Here, we report that CAR tonic signaling is caused by the intrinsic instability of the mAb single-chain variable fragment (scFv) to promote self-aggregation and signaling via the CD3ζ chain incorporated into the CAR construct. This phenomenon was detected in a CAR encoding either CD28 or 4-1BB costimulatory endodomains. Instability of the scFv was caused by specific amino acids within the framework regions (FWR) that can be identified by computational modeling. Substitutions of the amino acids causing instability, or humanization of the FWRs, corrected tonic signaling of the CAR, without modifying antigen specificity, and enhanced the antitumor effects of CAR-T cells. Overall, we demonstrated that tonic signaling of CAR-T cells is determined by the molecular instability of the scFv and that computational analyses of the scFv can be implemented to correct the scFv instability in CAR-T cells with either CD28 or 4-1BB costimulation.
Authors: Elisa Landoni, Giovanni Fucá, Jian Wang, Venkat R. Chirasani, Zhiyuan Yao, Elena Dukhovlinova, Soldano Ferrone, Barbara Savoldo, Lee K. Hong, Peishun Shou, Silvia Musio, Francesco Padelli, Gaetano Finocchiaro, Miriam Droste, Brian Kuhlman, Abdijapar Shamshiev, Serena Pellegatta, Nikolay V. Dokholyan, Gianpietro Dotti
Language: English
Topic: CAR-T cell therapy research
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Awards: STRATEGIES TO COUNTERACT RESISTANCE MECHANISMS IN CAR+ T-CELL-BASED IMMUNOTHERAPY FOR TRIPLE-NEGATIVE BREAST CANCER, T cell plasticity, fusion proteins and CAR T cell-based immunotherapy of head and neck cancer, G4119534169, G5850682332, IL-15 TRiKES-based specific immunotherapy of TNBC; resistance mechanisms, G7366156696, G9000033364, G9000191530, Nanoscale programming of celluar and physiological phenotypes, Penn State Clinical and Translational Science Institute, Computational Design of Protein Structures and Complexes, G958207224
OpenAlex ID: W4362525376