TY  - JOUR
A1  - Rydzek, Julian
A1  - Nerreter, Thomas
A1  - Peng, Haiyong
A1  - Jutz, Sabrina
A1  - Leitner, Judith
A1  - Steinberger, Peter
A1  - Einsele, Hermann
A1  - Rader, Christoph
A1  - Hudecek, Michael
T1  - Chimeric Antigen Receptor Library Screening Using a Novel NF-kappa B/NFAT Reporter Cell Platform
T2  - Molecular Therapy
N2  - Chimeric antigen receptor (CAR)-T cell immunotherapy is under intense preclinical and clinical investigation, and it involves a rapidly increasing portfolio of novel target antigens and CAR designs. We established a platform that enables rapid and high-throughput CAR-screening campaigns with reporter cells derived from the T cell lymphoma line Jurkat. Reporter cells were equipped with nuclear factor kappa B (NF kappa B) and nuclear factor of activated T cells (NFAT) reporter genes that generate a duplex output of enhanced CFP (ECFP) and EGFP, respectively. As a proof of concept, we modified reporter cells with CD19-specific and ROR1-specific CARs, and we detected high-level reporter signals that allowed distinguishing functional from non-functional CAR constructs. The reporter data were highly reproducible, and the time required for completing each testing campaign was substantially shorter with reporter cells (6 days) compared to primary CAR-T cells (21 days). We challenged the reporter platform to a large-scale screening campaign on a ROR1-CAR library, and we showed that reporter cells retrieved a functional CAR variant that was present with a frequency of only 6 in 1.05 x 10(6). The data illustrate the potential to implement this reporter platform into the preclinical development path of novel CAR-T cell products and to inform and accelerate the selection of lead CAR candidates for clinical translation.
KW  - NF-κB/NFAT reporter cells
KW  - chimeric antigen receptor
KW  - library screening
KW  - cancer immunotherapy
KW  - ROR1
Y1  - 2019
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/22719
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-227193
VL  - 27
IS  - 2
ER  -