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Methods to Evaluate the Antitumor Activity of Immune Checkpoint Inhibitors in Preclinical Studies
Bertrand Allard1,2, David Allard1,2, John Stagg3,4
1Centre de Recherche du Centre Hospitalier de l'Université de Montréal (CRCHUM), Institut du Cancer de Montréal, 900 Rue Saint-Denis, 10ième étage, Montréal, QC, Canada, H2X0A9.
Abstract:
Immune checkpoint inhibitors (ICI) are a new class of drugs characterized by their ability to enhance antitumor immune responses through the blockade of critical cell surface receptors involved in the maintenance of peripheral tolerance. The recent approval of ICI targeting CTLA-4 or PD-1 for the treatment of cancer constitutes a major breakthrough in the field of oncology and demonstrates the potential of immune-mediated therapies in achieving durable cancer remissions. The identification of new immune regulatory pathways that could be targeted to reactivate or boost antitumor immunity is now a very active field of research. In this context, the use of syngeneic mouse models and immune monitoring techniques are the cornerstone of proof-of-concept studies. In this chapter, we describe the general methodology to evaluate antitumor activity of ICI in immunocompetent mice. We outline protocols to reliably establish tumors in mice and generate lung metastasis through tail vein injections with the aim of testing the efficacy of ICI. We also present methods to analyze the composition of the tumor immune-infiltrate by multicolor flow cytometry.
Insights
Immune checkpoint inhibitors (ICI) enhance antitumor immunity by blocking immune-suppressing receptors. This chapter details methods for evaluating ICI efficacy in mouse models, crucial for developing new cancer immunotherapies.
Area of Science:
- Immunology
- Oncology
- Pharmacology
Background:
- Immune checkpoint inhibitors (ICI) represent a novel therapeutic class that enhances antitumor immune responses.
- ICI function by blocking cell surface receptors critical for maintaining peripheral tolerance, leading to improved cancer treatment outcomes.
- The development of durable cancer remissions through immune-mediated therapies marks a significant advancement in oncology.
Purpose of the Study:
- To describe methodologies for evaluating the antitumor activity of immune checkpoint inhibitors (ICI) in immunocompetent mouse models.
- To outline protocols for establishing tumors and lung metastases in mice for ICI efficacy testing.
- To present methods for analyzing the tumor immune-infiltrate composition using multicolor flow cytometry.
Main Methods:
- Utilizing syngeneic mouse models to assess the efficacy of ICI in vivo.
- Implementing protocols for tumor induction and the generation of lung metastases via tail vein injections.
- Employing multicolor flow cytometry for detailed analysis of immune cell populations within the tumor microenvironment.
Main Results:
- Established reliable protocols for evaluating ICI antitumor activity in preclinical mouse models.
- Demonstrated methods for generating and quantifying lung metastases to assess ICI efficacy.
- Provided a framework for immune monitoring, including the analysis of tumor-infiltrating lymphocytes.
Conclusions:
- Syngeneic mouse models and immune monitoring are essential for proof-of-concept studies of ICI.
- The described methodologies facilitate the rigorous evaluation of novel ICI therapies.
- This work supports the ongoing research into reactivating antitumor immunity for cancer treatment.
