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Published on: June 19, 2019
Microscale functional cytomics for studying hematologic cancers
Edmond W K Young1, Chorom Pak, Brad S Kahl
1Department of Biomedical Engineering, University of Wisconsin-Madison, 53705, USA.
Abstract:
An important problem in translational cancer research is our limited ability to functionally characterize behaviors of primary patient cancer cells and associated stromal cell types, and relate mechanistic understanding to therapy selection. Functional analyses of primary samples face at least 3 major challenges: limited availability of primary samples for testing, paucity of functional information extracted from samples, and lack of functional methods accessible to many researchers. We developed a microscale cell culture platform that overcomes these limitations, especially for hematologic cancers. A key feature of the platform is the ability to compartmentalize small populations of adherent and nonadherent cells in controlled microenvironments that can better reflect physiological conditions and enable cell-cell interaction studies. Custom image analysis was developed to measure cell viability and protein subcellular localizations in single cells to provide insights into heterogeneity of cellular responses. We validated our platform by assessing viability and nuclear translocations of NF-κB and STAT3 in multiple myeloma cells exposed to different conditions, including cocultured bone marrow stromal cells. We further assessed its utility by analyzing NF-κB activation in a primary chronic lymphocytic leukemia patient sample. Our platform can be applied to myriad biological questions, enabling high-content functional cytomics of primary hematologic malignancies.
Insights
Researchers developed a novel microscale cell culture platform for functional cytomics of primary cancer cells. This tool overcomes sample limitations, enabling detailed analysis of hematologic malignancies for improved therapy selection.
Area of Science:
- Oncology
- Biotechnology
- Cell Biology
Background:
- Translational cancer research faces challenges in functionally characterizing primary patient cancer cells and stromal cells.
- Limited sample availability, insufficient functional data, and inaccessible methods hinder progress in understanding cancer biology and selecting therapies.
Purpose of the Study:
- To develop and validate a microscale cell culture platform to overcome limitations in functional characterization of primary cancer cells.
- To enable high-content functional cytomics for hematologic malignancies, facilitating mechanistic understanding and therapy selection.
Main Methods:
- A microscale cell culture platform was designed to compartmentalize small cell populations in controlled microenvironments.
- Custom image analysis software was developed to measure single-cell viability and protein subcellular localization.
- The platform was validated using multiple myeloma and chronic lymphocytic leukemia patient samples, assessing cell viability and key signaling pathway activation (NF-κB, STAT3).
Main Results:
- The platform successfully compartmentalized adherent and nonadherent cells, mimicking physiological conditions and enabling cell-cell interaction studies.
- Single-cell analysis provided insights into cellular response heterogeneity, including viability and nuclear translocations of NF-κB and STAT3.
- NF-κB activation was successfully analyzed in a primary chronic lymphocytic leukemia patient sample.
Conclusions:
- The developed microscale cell culture platform effectively addresses limitations in functional analysis of primary cancer cells, particularly for hematologic malignancies.
- This platform enables high-content functional cytomics, offering a powerful tool for investigating cancer biology and guiding therapeutic strategies.
