Novel FRET-based Immunological Synapse Biosensor for the Prediction of Chimeric Antigen Receptor-T Cell Function.

Autor: Lee HN; Department of Pharmacology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Medical Research Institute, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Lee S; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, Republic of Korea.; BK21 FOUR Biomedical Science Project, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Hong J; Department of Pharmacology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Yoo H; Medicinal Materials Research Center, Biomedical Research Division, Korea Institute of Science and Technology (KIST), Seoul, 02792, Republic of Korea., Jeong J; Department of Pharmacology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Kim YW; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, Republic of Korea., Shin HM; Medical Research Institute, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, Republic of Korea.; BK21 FOUR Biomedical Science Project, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Jang M; Medicinal Materials Research Center, Biomedical Research Division, Korea Institute of Science and Technology (KIST), Seoul, 02792, Republic of Korea.; Department of Converging Science and Technology, Kyung Hee University, Seoul, 02447, Republic of Korea., Lee CH; Department of Pharmacology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Medical Research Institute, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, Republic of Korea.; BK21 FOUR Biomedical Science Project, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Cancer Research Institute, Seoul National University College of Medicine, Seoul, 03080, South Korea., Kim HR; Medical Research Institute, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, Republic of Korea.; BK21 FOUR Biomedical Science Project, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea., Seong J; Department of Pharmacology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Medical Research Institute, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, Republic of Korea.; Cancer Research Institute, Seoul National University College of Medicine, Seoul, 03080, South Korea.
Jazyk: angličtina
Zdroj: Small methods [Small Methods] 2024 Sep 11, pp. e2401016. Date of Electronic Publication: 2024 Sep 11.
DOI: 10.1002/smtd.202401016
Abstrakt: Chimeric antigen receptor (CAR)-T cell therapy has revolutionized cancer treatment. CARs are activated at the immunological synapse (IS) when their single-chain variable fragment (scFv) domain engages with an antigen, allowing them to directly eliminate cancer cells. Here, an innovative IS biosensor based on fluorescence resonance energy transfer (FRET) for the real-time assessment of CAR-IS architecture and signaling competence is presented. Using this biosensor, scFv variants for mesothelin-targeting CARs and identified as a novel scFv with enhanced CAR-T cell functionality despite its lower affinity than the original screened. The original CAR promoted internalization and trogocytosis, disrupting stable IS formation and impairing functionality are further observed. These findings emphasize the importance of enhancing IS quality rather than maximizing scFv affinity for superior CAR-T cell responses. Therefore, the FRET-based IS biosensor is a powerful tool for predicting CAR-T cell function, enabling the efficient engineering of next-generation CARs with enhanced antitumor potency.
(© 2024 The Author(s). Small Methods published by Wiley‐VCH GmbH.)
Databáze: MEDLINE
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