Site-specific genome engineering of primary human natural killer cells for programmable anti-tumor function

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Natural killer (NK) cells are a promising cell type for adoptive cell therapies.
• Existing gene editing methods for NK cells are challenging and require improvement.
• Developing efficient genome engineering strategies is crucial for advancing NK cell-based therapies.

Purpose of the Study:

• To establish a robust and modular workflow for genome engineering in primary human NK cells.
• To enable precise gene editing and transgene delivery for enhanced NK cell function.
• To facilitate the development of next-generation NK cell therapies.

Main Methods:

• Combined CRISPR/Cas9 with Adeno-Associated Virus 6 (AAV6)-mediated transgene delivery.
• Developed a feeder-free workflow for genome engineering in primary human NK cells.
• Targeted chimeric antigen receptor (CAR) transgene integration into inhibitory NK receptor loci, including TIGIT.

Main Results:

• Achieved efficient, site-specific transgene integration at various loci in NK cells.
• Demonstrated tunable transgene expression based on integration site and promoter.
• Identified the TIGIT locus as optimal for strong CAR expression and enhanced anti-tumor activity.

Conclusions:

• The developed genome engineering framework provides a powerful tool for NK cell therapy development.
• This strategy allows for precise genetic modifications to overcome NK cell limitations and tumor barriers.
• The findings support the rational design of enhanced NK cell-based cancer immunotherapies.