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Updated: Mar 18, 2026
Micromanipulation of Circulating Tumor Cells for Downstream Molecular Analysis and Metastatic Potential Assessment
Published on: May 14, 2019
Integrated Analysis of Multiple Biomarkers from Circulating Tumor Cells Enabled by Exclusion-Based Analyte Isolation
Jamie M Sperger1, Lindsay N Strotman2, Allison Welsh3
1Department of Medicine, University of Wisconsin-Madison, Madison, Wisconsin.
Purpose:
There is a critical clinical need for new predictive and pharmacodynamic biomarkers that evaluate pathway activity in patients treated with targeted therapies. A microscale platform known as VERSA (versatile exclusion-based rare sample analysis) was developed to integrate readouts across protein, mRNA, and DNA in circulating tumor cells (CTC) for a comprehensive analysis of the androgen receptor (AR) signaling pathway.
Experimental Design:
Utilizing exclusion-based sample preparation principles, a handheld chip was developed to perform CTC capture, enumeration, quantification, and subcellular localization of proteins and extraction of mRNA and DNA. This technology was validated across integrated endpoints in cell lines and a cohort of patients with castrate-resistant prostate cancer (CRPC) treated with AR-targeted therapies and chemotherapies.
Results:
The VERSA was validated in cell lines to analyze AR protein expression, nuclear localization, and gene expression targets. When applied to a cohort of patients, radiographic progression was predicted by the presence of multiple AR splice variants and activity in the canonical AR signaling pathway. AR protein expression and nuclear localization identified phenotypic heterogeneity. Next-generation sequencing with the FoundationOne panel detected copy number changes and point mutations. Longitudinal analysis of CTCs identified acquisition of multiple AR variants during targeted treatments and chemotherapy.
Conclusions:
Complex mechanisms of resistance to AR-targeted therapies, across RNA, DNA, and protein endpoints, exist in patients with CRPC and can be quantified in CTCs. Interrogation of the AR signaling pathway revealed distinct patterns relevant to tumor progression and can serve as pharmacodynamic biomarkers for targeted therapies. Clin Cancer Res; 1-11. ©2016 AACR.
Insights
New VERSA technology analyzes circulating tumor cells (CTCs) to reveal resistance mechanisms in prostate cancer patients on targeted therapy, identifying key biomarkers for treatment response.
Area of Science:
- Oncology
- Biomarker Discovery
- Molecular Diagnostics
Background:
- Clinical need for predictive and pharmacodynamic biomarkers in targeted cancer therapy.
- Current limitations in evaluating pathway activity in patients.
Purpose of the Study:
- Develop and validate the VERSA platform for comprehensive analysis of the androgen receptor (AR) signaling pathway in circulating tumor cells (CTCs).
- Integrate protein, mRNA, and DNA readouts from CTCs for a holistic view of pathway activity.
Main Methods:
- Development of a microscale, exclusion-based platform (VERSA) for CTC capture, enumeration, and molecular analysis.
- Validation in cell lines and a cohort of castrate-resistant prostate cancer (CRPC) patients.
- Analysis of AR protein expression, localization, gene expression, and genetic alterations (copy number, mutations) in CTCs.
Main Results:
- VERSA successfully analyzed AR signaling in cell lines and CRPC patient CTCs.
- Radiographic progression was predicted by AR splice variants and pathway activity.
- Phenotypic heterogeneity identified through AR protein expression and nuclear localization.
- Longitudinal CTC analysis revealed acquisition of AR variants during treatment.
Conclusions:
- Complex resistance mechanisms to AR-targeted therapies in CRPC can be quantified in CTCs.
- Interrogation of the AR signaling pathway in CTCs provides pharmacodynamic biomarkers for targeted therapies.
- The VERSA platform enables comprehensive analysis of AR pathway activity and resistance mechanisms.
