Related Experiment Video
Updated: Jul 11, 2025
Microfluidic Co-culture of Renal Healthy and Tumor Epithelium to Model Kidney Cancer Progression
Published on: January 31, 2025
Therapeutic targeting of tumor spheroids in a 3D microphysiological renal cell carcinoma-on-a-chip system
Chris P Miller1, Megan Fung2, Carla A Jaeger-Ruckstuhl2
1Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA, United States; Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, United States.
Abstract:
Metastatic renal cell carcinoma (RCC) remains an incurable disease for most patients highlighting an urgent need for new treatments. However, the preclinical investigation of new therapies is limited by traditional two-dimensional (2D) cultures which do not recapitulate the properties of tumor cells within a collagen extracellular matrix (ECM), while human tumor xenografts are time-consuming, expensive and lack adaptive immune cells. We report a rapid and economical human microphysiological system ("RCC-on-a-chip") to investigate therapies targeting RCC spheroids in a 3D collagen ECM. We first demonstrate that culture of RCC cell lines A498 and RCC4 in a 3D collagen ECM more faithfully reproduces the gene expression program of primary RCC tumors compared to 2D culture. We next used bortezomib as a cytotoxin to develop automated quantification of dose-dependent tumor spheroid killing. We observed that viable RCC spheroids exhibited collective migration within the ECM and demonstrated that our 3D system can be used to identify compounds that inhibit spheroid collective migration without inducing cell death. Finally, we demonstrate the RCC-on-a-chip as a platform to model the trafficking of tumor-reactive T cells into the ECM and observed antigen-specific A498 spheroid killing by engineered human CD8+ T cells expressing an ROR1-specific chimeric antigen receptor. In summary, the phenotypic differences between the 3D versus 2D environments, rapid imaging-based readout, and the ability to carefully study the impact of individual variables with quantitative rigor will encourage adoption of the RCC-on-a-chip system for testing a wide range of emerging therapies for RCC.
Insights
A novel "RCC-on-a-chip" system using 3D collagen cultures better models metastatic renal cell carcinoma (RCC) than 2D methods. This platform enables rapid, economical testing of new RCC therapies, including those involving engineered T cells.
Area of Science:
- Oncology
- Biotechnology
- Biomedical Engineering
Background:
- Metastatic renal cell carcinoma (RCC) is largely incurable, necessitating novel therapeutic strategies.
- Traditional 2D cell cultures and animal xenografts have limitations in accurately modeling RCC tumor microenvironments and immune responses.
- There is a critical need for advanced preclinical models that recapitulate tumor cell behavior within a complex extracellular matrix (ECM).
Purpose of the Study:
- To develop and validate a rapid, economical human microphysiological system (RCC-on-a-chip) for investigating therapies against RCC.
- To compare the gene expression profiles of RCC cells in 3D collagen ECM versus 2D cultures.
- To establish a platform for assessing drug efficacy, spheroid migration, and immune cell interactions in a 3D RCC model.
Main Methods:
- Culture of RCC cell lines (A498, RCC4) in 3D collagen ECM within a microphysiological system.
- Gene expression analysis to compare 3D vs. 2D culture models.
- Automated, dose-dependent quantification of tumor spheroid killing using bortezomib.
- Modeling of tumor-reactive T cell trafficking and antigen-specific killing by engineered T cells.
Main Results:
- 3D collagen ECM culture of RCC cells more accurately reflects primary tumor gene expression than 2D culture.
- The RCC-on-a-chip system successfully quantified dose-dependent spheroid killing and identified compounds inhibiting collective migration.
- The platform demonstrated T cell trafficking into the ECM and antigen-specific killing of RCC spheroids by engineered CD8+ T cells.
Conclusions:
- The RCC-on-a-chip system provides a more biologically relevant 3D microenvironment for preclinical RCC therapy testing.
- This 3D model offers a rapid, quantitative, and economical platform for evaluating diverse therapeutic strategies for metastatic RCC.
- The system's ability to model immune cell interactions and drug effects supports its adoption for advancing RCC treatment research.
