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Discrimination and Characterization of Heterocellular Populations Using Quantitative Imaging Techniques
Published on: June 30, 2017
A Multiplexed Approach to Assess Small Cell Lung Cancer Subtype Heterogeneity in Primary and Patient-Derived Tumor
Jin Ng1,2, Emma Pan3, Alex Johnston3
1ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
Abstract:
Unlocking the heterogeneity of cancers is crucial for developing therapeutic approaches that effectively eradicate disease. As our understanding of markers specific to cancer subclones or subtypes expands, there is a growing demand for advanced technologies that enable the simultaneous investigation of multiple targets within an individual tumor sample. Indeed, multiplex approaches offer distinct benefits, particularly when tumor specimens are small and scarce. Here we describe the utility of two fluorescence-based multiplex approaches; fluorescent Western blots, and multiplex immunohistochemistry (Opal™) staining to interrogate heterogeneity, using small cell lung cancer as an example. Critically, the coupling of Opal™ staining with advanced image quantitation, permits the dissection of cancer cell phenotypes at a single cell level. These approaches can be applied to patient biopsies and/or patient-derived xenograft (PDX) models and serve as powerful methodologies for assessing tumor cell heterogeneity in response to therapy or between metastatic lesions across diverse tissue sites.
Insights
Investigating cancer heterogeneity requires advanced multiplexing techniques. Fluorescent Western blots and multiplex immunohistochemistry (Opal™) enable detailed analysis of tumor samples, aiding therapeutic development.
Area of Science:
- Oncology
- Biotechnology
- Molecular Biology
Background:
- Understanding cancer heterogeneity is key for effective treatment development.
- Expanding knowledge of cancer subclone and subtype markers necessitates advanced analytical technologies.
- Multiplex approaches are particularly beneficial for analyzing limited tumor samples.
Purpose of the Study:
- To demonstrate the utility of fluorescence-based multiplex approaches for interrogating cancer heterogeneity.
- To showcase the application of fluorescent Western blots and multiplex immunohistochemistry (Opal™) staining.
- To highlight the combined power of Opal™ staining and image quantitation for single-cell phenotype analysis.
Main Methods:
- Utilized fluorescent Western blots to analyze protein expression.
- Employed multiplex immunohistochemistry (Opal™) staining for spatial analysis of markers.
- Integrated advanced image quantitation with Opal™ staining for detailed cellular analysis.
- Applied these methods to small cell lung cancer (SCLC) models and patient biopsies.
Main Results:
- Demonstrated the capability of multiplex approaches to reveal cancer cell heterogeneity.
- Showcased the effectiveness of Opal™ staining coupled with image quantitation for single-cell phenotyping.
- Validated the utility of these methods in patient-derived xenograft (PDX) models and clinical samples.
Conclusions:
- Fluorescence-based multiplex approaches, including fluorescent Western blots and Opal™ staining, are powerful tools for dissecting tumor heterogeneity.
- The combination of Opal™ staining and advanced image quantitation allows for high-resolution analysis of cancer cell phenotypes.
- These methodologies are applicable to various sample types, including patient biopsies and PDX models, for assessing therapeutic response and metastatic heterogeneity.
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