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Updated: Dec 31, 2025
Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs
Published on: December 16, 2022
Evaluation of PD-1 blockade using a multicellular tumor spheroid model
Shujuan Zhou1, Mei Zhu1,2,3, Fanyan Meng1
1The Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University & Clinical Cancer Institute of Nanjing University Nanjing, China.
Abstract:
Three-dimensional multicellular tumor spheroids are invaluable due to their ability to recapitulate major physiological characteristics of solid tumors. The lack of appropriate in vitro models to evaluate the effects of PD-1 blockade has restricted use of PD-1 inhibitors. Here, we established spheroids from a human gastric cell line to evaluate the cytotoxicity of the PD-1 blockade. Mature spheroids were formed 72 h after seeding tumor cells into ultra-low attachment microplates. We induced PD-L1 expression of spheroids using IFN-γ and TNF-α. Confocal images showed that PD-1 blockade enhanced T-cell cytotoxicity against PD-L1-expressing spheroids. Mechanistic studies revealed that PD-1 blockade increased expression of multiple chemokines. Altogether, our study demonstrates that tumor spheroids provide a novel platform to evaluate the effects of PD-1 blockade and might be further utilized to develop promising combination therapies.
Insights
Three-dimensional tumor spheroids effectively model solid tumors for evaluating PD-1 blockade therapy. This study shows PD-1 blockade enhances T-cell cytotoxicity against these spheroids, offering a new platform for drug development.
Area of Science:
- Oncology
- Immunotherapy
- Biomedical Engineering
Background:
- Three-dimensional multicellular tumor spheroids mimic solid tumor physiology.
- Existing in vitro models are insufficient for evaluating PD-1 blockade efficacy.
- PD-1 inhibitors are crucial in cancer immunotherapy.
Purpose of the Study:
- To establish and utilize tumor spheroids as an in vitro model for assessing PD-1 blockade cytotoxicity.
- To investigate the effects of PD-1 blockade on T-cell-mediated killing of gastric cancer spheroids.
- To explore the underlying mechanisms of PD-1 blockade in a spheroid model.
Main Methods:
- Generation of mature multicellular tumor spheroids from a human gastric cell line using ultra-low attachment microplates.
- Induction of PD-L1 expression on spheroids using interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α).
- Assessment of T-cell cytotoxicity against PD-L1-expressing spheroids following PD-1 blockade using confocal microscopy and mechanistic studies.
Main Results:
- PD-1 blockade significantly enhanced T-cell-mediated cytotoxicity against PD-L1-expressing gastric cancer spheroids.
- PD-1 blockade treatment led to increased expression of multiple chemokines within the spheroid microenvironment.
- The study successfully validated tumor spheroids as a functional in vitro model for PD-1 blockade evaluation.
Conclusions:
- Tumor spheroids serve as a valuable and novel platform for evaluating the efficacy of PD-1 blockade therapies.
- The findings support the potential use of this spheroid model for developing and optimizing future cancer combination therapies.
- This model provides insights into the immunomodulatory effects of PD-1 blockade, including chemokine modulation.
