Comparison of lentiviral and sleeping beauty mediated αβ T cell receptor gene transfer

Area of Science:

• Immunology
• Gene Therapy
• Molecular Biology

Background:

• Current T cell receptor gene transfer relies on costly and inflexible retroviral or lentiviral systems.
• These systems are used for chimeric antigen receptors and T cell receptors, but have limitations.
• There is a need for more cost-effective and adaptable platforms for T cell therapy development.

Purpose of the Study:

• To evaluate the Sleeping Beauty transposon system as a cost-effective alternative for T cell receptor gene transfer.
• To assess the efficiency and safety of Sleeping Beauty transposon-mediated gene transfer for T cell therapies.
• To compare Sleeping Beauty transposon system with lentiviral vectors for T cell receptor gene delivery.

Main Methods:

• Utilized Sleeping Beauty transposon plasmid system with hyperactive SB100X transposase.
• Nucleofection was used for efficient gene transfer into primary T cells.
• Optimized murine-human chimeric T cell receptor specific for Wilm's tumour antigen was employed.
• Compared gene transfer efficiency, cell expansion, and in vitro/in vivo lysis with lentiviral transduction.

Main Results:

• Stable transposition of the T cell receptor gene was achieved using the Sleeping Beauty system.
• While transfer efficiency was lower than lentiviral vectors, cells were successfully enriched and expanded.
• The system mediated effective target cell lysis both in vitro and in vivo.
• Integration sites were randomly distributed, unlike the targeted integration of lentiviral vectors.

Conclusions:

• The Sleeping Beauty plasmid-based system is a flexible and adaptable platform for T cell receptor gene therapies.
• This system allows for accelerated early-phase assessment of T cell receptor gene therapies.
• The random integration profile offers a potential safety advantage over lentiviral vectors.