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Methyl-binding DNA capture Sequencing for Patient Tissues
Published on: October 31, 2016
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Analysis of DNA methylation in single circulating tumor cells
C F Pixberg1, K Raba2, F Müller1
1Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, Düsseldorf, Germany.
Oncogene
|January 10, 2017
Summary
This study developed a novel single-cell assay to analyze DNA methylation in circulating tumor cells (CTCs). The findings reveal epigenetic heterogeneity in CTCs, suggesting active epigenetic regulation of genes driving cancer metastasis.
Area of Science:
- Oncology
- Epigenetics
- Molecular Biology
Background:
- Direct analysis of circulating tumor cells (CTCs) provides insights into cancer's systemic spread.
- Epithelial-to-mesenchymal transition (EMT) is crucial for tumor cell dissemination and metastasis.
- Understanding epigenetic regulation of EMT-associated genes in CTCs is vital for cancer research.
Purpose of the Study:
- To investigate promoter methylation of EMT-regulating genes in single CTCs.
- To develop and validate a single-cell assay for multiplexed DNA methylation analysis.
- To explore epigenetic heterogeneity among CTCs from metastatic breast and prostate cancer patients.
Main Methods:
- Developed a single-cell agarose-embedded bisulfite treatment protocol for multiplexed DNA methylation analysis (multiplexed-scAEBS).
- Simultaneously analyzed methylation of miR-200c/141, miR-200b/a/429, and CDH1 genes in single cells.
- Applied the validated assay to 159 single CTCs from metastatic breast and castration-resistant prostate cancer patients.
Main Results:
- The multiplexed-scAEBS assay achieved 70% DNA amplification efficiency with accurate methylation patterns.
- CTCs exhibited methylation patterns more akin to epithelial-like cells than to white blood cells.
- Promoter methylation of the microRNA-200 family was significantly higher in prostate CTCs compared to breast CTCs.
- Single-cell analysis uncovered significant epigenetic heterogeneity within CTC populations.
Conclusions:
- The developed assay enables robust single-cell DNA methylation analysis of CTCs.
- Epigenetic heterogeneity in CTCs suggests active, tumor-specific epigenetic regulation during metastasis.
- Findings highlight the role of epigenetic modifications in the biology of circulating tumor cells.
