Chimeric Antigen Receptor Library Screening Using a Novel NF-κB/NFAT Reporter Cell Platform

Rydzek J, Nerreter T, Peng H, Jutz S, Leitner J, Steinberger P, Einsele H, Rader C, Hudecek M (2019)

Publication Type: Journal article

Publication year: 2019

Journal

Molecular Therapy Cell Press

Book Volume: 27

Pages Range: 287-299

Journal Issue: 2

DOI: 10.1016/j.ymthe.2018.11.015

Abstract

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 κB (NF-κB) 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 × 106. 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.

Involved external institutions

Universitätsklinikum Würzburg DE Germany (DE) Scripps Research Institute (TSRI) US United States (USA) (US) Medizinische Universität Wien AT Austria (AT)

How to cite

APA:

Rydzek, J., Nerreter, T., Peng, H., Jutz, S., Leitner, J., Steinberger, P.,... Hudecek, M. (2019). Chimeric Antigen Receptor Library Screening Using a Novel NF-κB/NFAT Reporter Cell Platform. Molecular Therapy, 27(2), 287-299. https://doi.org/10.1016/j.ymthe.2018.11.015

MLA:

Rydzek, Julian, et al. "Chimeric Antigen Receptor Library Screening Using a Novel NF-κB/NFAT Reporter Cell Platform." Molecular Therapy 27.2 (2019): 287-299.

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