Evaluating Multimodal Functionality of Chimeric Antigen Receptor T Cells at Single-Cell Resolution through...

Area of Science:

• Immunology
• Cell Therapy
• Biotechnology

Background:

• Adoptive T-cell therapy, specifically chimeric antigen receptor (CAR)-T cell therapy, shows promise for hematological malignancies.
• Variability in clinical outcomes is observed, with polyfunctional T cells correlating with durable remission.
• Current CAR-T cell product evaluations use bulk measurements, hindering the study of functional subpopulations.

Purpose of the Study:

• To develop and demonstrate a workflow for evaluating individual CAR-T cell functionality.
• To enable the isolation of highly functional CAR-T cell subpopulations for next-generation therapy design.

Main Methods:

• Utilized an optofluidics platform to assess individual CAR-T cell cytokine secretion and CD137 expression.
• Integrated cytotoxic activity assessment as an optional component.
• Enabled isolation of CAR-T cells with multimodal functionality.

Main Results:

• Demonstrated a workflow for single-cell analysis of CAR-T cell function, including cytokine profiles and activation markers.
• Showcased the ability to identify and isolate CAR-T cells with superior polyfunctional characteristics.
• Established a method to link pre-infusion CAR-T cell quality to potential therapeutic efficacy.

Conclusions:

• The optofluidics platform provides a powerful tool for detailed characterization of CAR-T cell products.
• Isolation of highly functional CAR-T cells can inform the development of more effective and durable cancer immunotherapies.
• This approach facilitates the design of next-generation CAR-T cell therapies with improved clinical outcomes.