Protocol to prepare functional cellular nanovesicles with PD1 and TRAIL to boost antitumor response

Area of Science:

• Biomedical Engineering
• Cancer Research
• Immunology

Background:

• Immunotherapy shows promise in cancer treatment but is limited by tumor microenvironment immunosuppression.
• Multiple immunosuppressive pathways hinder the effectiveness of current cancer immunotherapies.
• A need exists for novel strategies to overcome tumor-induced immune suppression.

Purpose of the Study:

• To present a protocol for preparing functional cellular nanovesicles engineered to enhance cancer immunotherapy.
• To develop nanovesicles displaying Programmed Death 1 (PD1) and TNF-Related Apoptosis-Inducing Ligand (TRAIL).
• To investigate the potential of these nanovesicles in overcoming tumor immunosuppression.

Main Methods:

• Utilized HEK293-FT cells for the production of cellular nanovesicles.
• Engineered nanovesicles to display PD1, a checkpoint inhibitor, and TRAIL, a molecule inducing immunogenic cell death.
• Developed a step-by-step protocol for nanovesicle preparation and characterization.

Main Results:

• Successfully prepared functional cellular nanovesicles displaying both PD1 and TRAIL.
• TRAIL component is designed to induce immunogenic cancer cell death, initiating an immune response.
• PD1 component aims to block the PD1/PDL1 checkpoint, reactivating tumor-specific CD8+ T cells.

Conclusions:

• The developed protocol provides a method for generating novel cellular nanovesicles for cancer immunotherapy.
• These engineered nanovesicles hold potential for overcoming tumor microenvironment-mediated immunosuppression.
• This approach may broaden the applicability of immunotherapy to a larger patient population.