Intein-based modular chimeric antigen receptor platform for specific CD19/CD20 co-targeting.

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• Chimeric antigen receptor (CAR) T-cell therapy has transformed B-cell malignancy treatment.
• Challenges like antigen escape and tumor heterogeneity limit CAR T-cell efficacy.
• Modular CARs offer a strategy to overcome these limitations by targeting multiple antigens.

Purpose of the Study:

• To introduce CARtein, a novel modular CAR platform utilizing intein interactions.
• To enable simultaneous targeting of CD19 and CD20 antigens with a universal CAR backbone.
• To establish a foundation for a versatile multi-antigen targeting system.

Main Methods:

• Development of the CARtein platform with a universal CAR signaling backbone and scFv-intein recognition partners.
• Validation of CARtein functionality using CD19 and/or CD20 expressing Raji and K562 cells.
• Assessment of T-cell activation via NFAT and NFκB promoter activity and CD69 expression.

Main Results:

• The CARtein system successfully generated fully active CARs.
• Significant T-cell activation was observed when targeting cells expressing CD19 and/or CD20.
• Demonstrated specific activation through covalent binding of scFv-intein partners to the CAR backbone.

Conclusions:

• CARtein provides a new modular CAR platform for multi-antigen targeting in B-cell malignancies.
• The system utilizes intein-mediated covalent binding for precise CAR activation.
• This approach holds promise for overcoming antigen escape and improving CAR T-cell therapy outcomes.