A 3D multi-compartment assembloid to study combined immune cell infiltration and cytotoxicity

Area of Science:

• Immunology
• Cancer Biology
• Biotechnology

Background:

• Immune cell-mediated cancer cell killing involves complex infiltration into solid tumors.
• Traditional 2D assays do not fully capture 3D immune cell-extracellular matrix (ECM) interactions crucial for tumor infiltration.
• Understanding these interactions is vital for developing effective immunotherapies.

Purpose of the Study:

• To develop and validate a novel 3D multi-compartment assembloid assay for studying immune cell infiltration and cytotoxicity within a tumor microenvironment.
• To compare the performance of this 3D assay against standard 2D cytotoxicity assays.
• To provide a tool for mechanistic studies of immune cell-tumor microenvironment interactions.

Main Methods:

• Development of a 3D multi-compartment assembloid model.
• Co-culture of immune cells (including chimeric antigen receptor (CAR) T cells) with tumor cells within the 3D matrix.
• Comparison with standard 2D cytotoxicity assays.
• Utilizing various imaging techniques for observation and quantification.

Main Results:

• The 3D assembloid assay successfully models immune cell stromal invasion, matrix migration, tumor infiltration, and subsequent cytotoxicity.
• Direct observation and quantification of each infiltration stage were achieved in the 3D model.
• The 3D assay provides a more comprehensive view of immune cell-ECM-tumor interactions compared to 2D assays.

Conclusions:

• The developed 3D infiltration/cytotoxicity assay is a valuable tool for mechanistic studies of anti-tumor immunity.
• This assay enhances the understanding of immune cell interactions within the complex tumor microenvironment.
• It offers a more physiologically relevant platform for evaluating immunotherapies, including CAR T cell therapies.