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

Area of Science:

• Immunotherapy
• Cellular Engineering
• Cancer Research

Background:

• Chimeric antigen receptor (CAR)-T cell immunotherapy is a rapidly advancing field with numerous target antigens and CAR designs.
• Efficient screening of novel CAR constructs is crucial for preclinical development and clinical translation.

Purpose of the Study:

• To establish a high-throughput reporter cell platform for rapid CAR screening.
• To demonstrate the platform's ability to identify functional CAR variants efficiently.

Main Methods:

• Development of Jurkat T cell reporter lines expressing NF-κB and NFAT reporter genes (ECFP and EGFP).
• Modification of reporter cells with CD19- and ROR1-specific CARs for proof-of-concept validation.
• Large-scale screening of a ROR1-CAR library using the reporter platform.

Main Results:

• The reporter platform demonstrated high reproducibility and significantly reduced testing time (6 days vs. 21 days for primary CAR-T cells).
• Functional CAR constructs were clearly distinguished from non-functional ones via reporter signals.
• A rare functional ROR1-CAR variant was successfully identified from a large library.

Conclusions:

• The reporter platform offers a robust and accelerated method for CAR-T cell preclinical development.
• This technology can significantly inform and expedite the selection of lead CAR candidates for clinical application.