Protocol for assessing pharmacokinetics and pharmacodynamics of human CAR-NKT cells in humanized mouse models using...

Area of Science:

• Immunology
• Cell Therapy
• Preclinical Models

Background:

• Invariant natural killer T (NKT) cells are crucial immune cells with potent anti-tumor capabilities, bridging innate and adaptive immunity.
• Chimeric antigen receptor (CAR)-engineered NKT cells derived from peripheral blood mononuclear cells (PBMCs) demonstrate significant anti-tumor activity.
• Developing effective CAR-NKT cell therapies requires robust methods to evaluate their in vivo behavior.

Purpose of the Study:

• To establish and present a standardized protocol for assessing the pharmacokinetics and pharmacodynamics (PK/PD) of CAR-NKT cells.
• To detail the application of in vivo bioluminescence imaging (BLI) for tracking CAR-NKT cell behavior in preclinical models.
• To support the advancement of CAR-NKT cell-based immunotherapies through reliable PK/PD assessment.

Main Methods:

• Utilizing humanized mouse models, both tumor-free and tumor-bearing.
• Implementing in vivo bioluminescence imaging (BLI) to monitor CAR-NKT cell distribution and persistence.
• Describing procedures for evaluating CAR-NKT cell infiltration into tumor sites.

Main Results:

• The protocol enables quantitative assessment of CAR-NKT cell distribution and persistence over time.
• Bioluminescence imaging effectively visualizes CAR-NKT cell localization within different tissues and tumor microenvironments.
• The methodology provides critical data for understanding the in vivo fate of CAR-NKT cells.

Conclusions:

• This protocol offers a valuable tool for preclinical evaluation of CAR-NKT cell therapies.
• The described BLI approach facilitates comprehensive PK/PD analysis of CAR-NKT cells in relevant models.
• Accurate assessment of CAR-NKT cell behavior is essential for optimizing their therapeutic efficacy and safety.