Identification and Characterization of Fully Human FOLR1-Targeting CAR T Cells for the Treatment of Ovarian Cancer

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• Chimeric Antigen Receptor (CAR) T-cell therapy shows promise for hematological malignancies.
• Challenges like immunogenicity, toxicity, and lack of comprehensive evaluation workflows hinder CAR T-cell therapeutic potential.

Purpose of the Study:

• To establish a stringent, multistep workflow for thorough evaluation and selection of lead CAR T-cell candidates.
• To characterize a human FOLR1-directed CAR binding domain, focusing on specificity.

Main Methods:

• Implemented a multistep workflow involving extensive specificity assessments using flow cytometry and imaging.
• Utilized high-throughput screening and advanced in vitro assays to evaluate CAR T-cell functionality and specificity.
• Determined on-/off-target and off-tumor reactivity of CAR binding domains.

Main Results:

• Characterized a human FOLR1-directed CAR binding domain with detailed specificity profiling.
• Demonstrated that comprehensive assays for CAR functionality and binding specificity are complementary.
• Successfully identified critical specificity considerations early in the CAR development process.

Conclusions:

• The developed workflow enables early identification of critical specificity considerations for CAR T-cell therapies.
• This approach aims to overcome limitations related to immunogenicity and toxicity, improving the selection of effective CAR T-cell candidates.
• This strategy supports the advancement of future CAR T-cell therapies by ensuring robust candidate assessment.