A Novel TumorGraft3D immune co-culture platform for high throughput immune-oncology screening
The complexity of the tumor immune microenvironment (TIME) plays a critical role in immunotherapy treatment response, and this requires robust and faithful preclinical research models recapitulating patient-specific tumor-immune interactions1. Immune oncology research has generated an attractive potential to model TIME complexity at the level of a single patient. Rapid progress has been made, yet there are still challenges while developing rigorous ex-vivo models mimicking the hierarchical architecture and cellular synergy within TIME.
Although patient-derived organoid technology overcomes the limitation of standard 2D methods allowing for proliferating cells and high throughput drug screening, it still lacks functional immune components to successfully test immune-checkpoint inhibitors (ICIs) in reproducible, and cost-effective system.
The challenge of replicating immunity in vitro relies on immune cells rapidly losing viability and functional features and the availability of autologous patient immune cells. Providing a solution to this challenge Champions Oncology leverages the patient-derived xenograft bank generating co-culture embedding organoid and immune cells from the same patient in autologous settings.
Champions Oncology’s novel and versatile TumorGraft3D co-culture platform utilizes organoids from patient-derived xenograft models with matching immune cells. The characterization of the platform showed the proliferation of the cancer cells and functional immune profiling by flow cytometry. The molecular profile of the organoids composing the platform showed a tight overlap with the original tumor highlighting the clinical relevance of the model.
To establish autologous Tumorgraft3D co-cultures, surgical resections from patients were used to generate patient-derived xenograft organoids, and TILs were expanded using standard Rapid Expansion protocol2. 3D confocal analysis allowed for a deep evaluation of cancer-immune cellular interaction at the single-cell level. We developed a staining system allowing us to differentiate cancer-immune cell spatial distribution and cellular death. Our proprietary pipeline of image analysis allowed the evaluation of tumor infiltration and drug cytotoxicity.
The novel co-culture platform has been tested with various indications allowing to test ICIs drug response including pembrolizumab, nivolumab, durvalumab, and ipilimumab. The quantified image analysis results demonstrate varying degrees of TIL infiltration and cytotoxicity with different ICIs and our findings corroborate emerging clinical results.
In summary, this novel platform allows high throughput screening of novel immune targeting agents to determine impacts on immune cell infiltration and tumor cytotoxicity. TumorGraft3D is versatile and can be used to test a variety of therapeutic modalities, alone or in combination, including but not limited to allogeneic, innate cell therapies, engineered immune cells as well as therapeutics targeting proliferation mechanisms.