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Analysis of Antitumor Effects of CAR-T Cells in Mice with Solid Tumors
Alba Rodriguez-Garcia1, Keisuke Watanabe2, Sonia Guedan3
1Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Abstract:
Animal models provide an essential tool to study the efficacy of CAR-T cell treatments. Most of the current works test human CAR-T cells in immunodeficient animals, typically NOD Scid Gamma (NSG) mice transplanted with human tumors. Despite the limitations of this model, including the difficulty to study the interaction between CAR-T cells and the human innate system and to assess the toxicity of this therapy, NSG are extensively used for adoptive T cell transfer studies. In this chapter, we will describe the protocols to test CAR-T cells in NSG animals with solid tumors. We first describe the implantation of human xenograft tumors in NSG animals, followed by CAR-T cell administration and assessment of antitumor responses. We will also review the protocols to analyze T cell persistence in the blood of treated animals. Finally, we will focus on the analysis of the tumors at the end point of the experiment, including the percentage, phenotype, and function of tumor infiltrating T cells, and loss of antigen expression.
Insights
This chapter details protocols for testing chimeric antigen receptor T-cell (CAR-T) therapies in NOD Scid Gamma (NSG) mice with solid tumors. It covers tumor implantation, CAR-T cell administration, and analysis of antitumor responses and T-cell persistence.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Chimeric antigen receptor T-cell (CAR-T) therapy shows promise in cancer treatment.
- Animal models are crucial for evaluating CAR-T cell efficacy and safety.
- NOD Scid Gamma (NSG) mice are commonly used for xenograft studies despite limitations in modeling human innate immune interactions.
Purpose of the Study:
- To describe standardized protocols for assessing CAR-T cell efficacy in NSG mice bearing solid tumors.
- To outline methods for evaluating antitumor responses, T-cell persistence, and tumor-infiltrating lymphocyte characteristics.
Main Methods:
- Human tumor xenograft implantation in NSG mice.
- Administration of CAR-T cells and monitoring of antitumor effects.
- Analysis of T-cell persistence in peripheral blood.
- Endpoint tumor analysis including T-cell infiltration, phenotype, function, and antigen expression.
Main Results:
- Established protocols for CAR-T cell administration and efficacy assessment in a solid tumor NSG mouse model.
- Methods for quantifying T-cell persistence and characterizing tumor-infiltrating lymphocytes.
- Assessment of potential mechanisms of resistance, such as antigen loss.
Conclusions:
- Standardized protocols enable robust evaluation of CAR-T cell therapy in preclinical solid tumor models.
- Comprehensive analysis provides insights into CAR-T cell activity, persistence, and tumor microenvironment interactions.
- This approach facilitates the study of CAR-T cell efficacy and potential resistance mechanisms in vivo.
