A method for polyclonal antigen-specific T cell-targeted genome editing (TarGET) for adoptive cell transfer...

Area of Science:

• Immunology
• Cell Therapy
• Genomic Engineering

Background:

• Adoptive cell therapy using antigen-specific T cells is crucial for immunocompromised patients facing viral infections.
• Tracking infused T cells *in vivo* is essential for evaluating therapeutic efficacy but current genetic labeling methods lack specificity.

Purpose of the Study:

• To develop a specific method for tracking donor-derived, antigen-specific T cells post-infusion.
• To enable lineage tracing and monitoring of T cell expansion and function in adoptive cell therapy.

Main Methods:

• CRISPR-targeted genome integration of a barcoded gene into Epstein-Barr virus-stimulated T cells.
• Single-cell and deep sequencing for lineage tracing and gene expression analysis.
• Assessment of T cell enrichment and *in vitro* cytotoxicity against target cells.

Main Results:

• Successfully demonstrated CRISPR-mediated barcoding for exclusive identification of expanded virus-specific T cell lineages.
• Showcased improved enrichment of antigen-specific T cells.
• Revealed expansion profiles of specific T cell clones and their gene expression signatures through lineage tracing.

Conclusions:

• The developed barcoding method enhances traceability and monitoring of T cells in adoptive cell therapy.
• This approach holds potential for optimizing immunotherapeutic regimens by providing detailed insights into T cell behavior.
• Facilitates precise evaluation of therapeutic T cell clones and their functional outcomes.