Related Experiment Video
Updated: Feb 19, 2026
High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids
Published on: June 14, 2021
Ex Vivo Profiling of PD-1 Blockade Using Organotypic Tumor Spheroids
Russell W Jenkins1,2, Amir R Aref1,3, Patrick H Lizotte1,3
1Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts.
Abstract:
Ex vivo systems that incorporate features of the tumor microenvironment and model the dynamic response to immune checkpoint blockade (ICB) may facilitate efforts in precision immuno-oncology and the development of effective combination therapies. Here, we demonstrate the ability to interrogate ex vivo response to ICB using murine- and patient-derived organotypic tumor spheroids (MDOTS/PDOTS). MDOTS/PDOTS isolated from mouse and human tumors retain autologous lymphoid and myeloid cell populations and respond to ICB in short-term three-dimensional microfluidic culture. Response and resistance to ICB was recapitulated using MDOTS derived from established immunocompetent mouse tumor models. MDOTS profiling demonstrated that TBK1/IKKε inhibition enhanced response to PD-1 blockade, which effectively predicted tumor response in vivo Systematic profiling of secreted cytokines in PDOTS captured key features associated with response and resistance to PD-1 blockade. Thus, MDOTS/PDOTS profiling represents a novel platform to evaluate ICB using established murine models as well as clinically relevant patient specimens.Significance: Resistance to PD-1 blockade remains a challenge for many patients, and biomarkers to guide treatment are lacking. Here, we demonstrate feasibility of ex vivo profiling of PD-1 blockade to interrogate the tumor immune microenvironment, develop therapeutic combinations, and facilitate precision immuno-oncology efforts. Cancer Discov; 8(2); 196-215. ©2017 AACR.See related commentary by Balko and Sosman, p. 143See related article by Deng et al., p. 216This article is highlighted in the In This Issue feature, p. 127.
Insights
This study introduces organotypic tumor spheroids for ex vivo immune checkpoint blockade (ICB) testing. This novel platform accurately models tumor response and aids in developing precision immuno-oncology therapies.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Immune checkpoint blockade (ICB) therapy shows promise but faces challenges with patient response and resistance.
- Developing predictive biomarkers and combination therapies is crucial for advancing precision immuno-oncology.
Purpose of the Study:
- To establish and validate an ex vivo platform for modeling tumor response to ICB.
- To investigate novel therapeutic combinations and identify biomarkers for ICB response.
Main Methods:
- Utilized murine- and patient-derived organotypic tumor spheroids (MDOTS/PDOTS) in 3D microfluidic culture.
- Assessed ICB response, including PD-1 blockade, and evaluated the effect of TBK1/IKKε inhibition.
- Systematically profiled secreted cytokines from PDOTS to identify response and resistance signatures.
Main Results:
- MDOTS/PDOTS successfully recapitulated ICB response and resistance observed in vivo.
- TBK1/IKKε inhibition was shown to enhance PD-1 blockade response, predicting in vivo tumor outcomes.
- Cytokine profiling of PDOTS identified key features associated with ICB response and resistance.
Conclusions:
- MDOTS/PDOTS profiling offers a novel and effective platform for evaluating ICB in both preclinical models and patient specimens.
- This approach facilitates the development of combination therapies and advances precision immuno-oncology efforts by interrogating the tumor immune microenvironment.
