Efficient NK cell transduction with VSV-G-pseudotyped lentiviral vectors

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Chimeric antigen receptor (CAR)-NK cell therapy offers potential for off-the-shelf cancer treatment, overcoming limitations of CAR-T cell therapies.
• Lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein G (VSV-G) are common for cell modification but show low transduction efficiency in NK cells.

Purpose of the Study:

• To investigate and optimize the factors influencing lentiviral transduction efficiency in Natural Killer (NK) cells.
• To develop a robust workflow for efficient genetic modification of NK cells for therapeutic applications.

Main Methods:

• Exploration of NK cell activation strategies using interleukins.
• Optimization of CAR construct design and lentivirus pseudotype selection.
• Evaluation of transduction enhancers, including BX795 and retronectin, for enhancing VSV-G lentiviral vector transduction.

Main Results:

• Achieved high transduction efficiency (91% GFP, 80% CAR) using an optimized workflow involving IL-activated NK cells, a specific CAR construct, and VSV-G pseudotyped lentiviral vectors with BX795 and retronectin.
• Demonstrated that optimized transduction conditions do not compromise NK cell phenotype, growth, or cytotoxic function.
• Validated the use of VSV-G pseudotyped lentiviral vectors for efficient therapeutic NK cell production.

Conclusions:

• The study presents an optimized protocol for efficient and safe lentiviral transduction of NK cells.
• This workflow facilitates the production of CAR-NK cell products using established lentiviral vector technology with a strong safety profile.
• The findings support the advancement of allogeneic CAR-NK cell therapies for cancer treatment.