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Published on: July 5, 2019
Head and Neck Cancer Stem Cell-Enriched Spheroid Model for Anticancer Compound Screening
Larisa Goričan1, Boris Gole1, Uroš Potočnik1,2
1Centre for Human Molecular Genetics and Pharmacogenomics, Faculty of Medicine, University of Maribor, SI-2000 Maribor, Slovenia.
Abstract:
Cancer stem cells (CSCs), a rare cell population in tumors, are resistant to conventional chemotherapy and thus responsible for tumor recurrence. To screen for active compounds targeting CSCs, a good CSC-enriched model compatible with high-throughput screening (HTS) is needed. Here, we describe a new head and neck cancer stem cell-enriched spheroid model (SCESM) suitable for HTS analyses of anti-CSC compounds. We used FaDu cells, round-bottom ultra-low adherent (ULA) microplates, and stem medium. The formed spheroids displayed increased expression of all stem markers tested (qRT-PCR and protein analysis) in comparison to the FaDu cells grown in a standard adherent culture or in a well-known HTS-compatible multi-cellular tumor spheroid model (MCTS). Consistent with increased stemness of the cells in the spheroid, confocal microscopy detected fast proliferating cells only at the outer rim of the SCESM spheroids, with poorly/non-proliferating cells deeper in. To confirm the sensitivity of our model, we used ATRA treatment, which strongly reduced the expression of selected stem markers. Altogether, we developed a CSC-enriched spheroid model with a simple protocol, a microplate format compatible with multimodal detection systems, and a high detection signal, making it suitable for anti-CSC compounds' HTS.
Insights
Researchers developed a novel head and neck cancer stem cell-enriched spheroid model (SCESM) for high-throughput screening (HTS). This model effectively identifies compounds targeting cancer stem cells (CSCs) and aids in overcoming tumor recurrence.
Area of Science:
- Oncology
- Stem Cell Biology
- Drug Discovery
Background:
- Cancer stem cells (CSCs) drive tumor recurrence due to chemoresistance.
- Effective high-throughput screening (HTS) models are crucial for identifying anti-CSC compounds.
- Existing models may not be optimal for CSC-enriched screening.
Purpose of the Study:
- To develop and validate a novel CSC-enriched spheroid model for HTS of anti-CSC compounds.
- To establish a model suitable for head and neck cancers.
- To facilitate the discovery of novel therapeutic strategies against CSCs.
Main Methods:
- Utilized FaDu cells in ultra-low adherent (ULA) microplates with stem medium to form spheroids.
- Characterized spheroid stemness via qRT-PCR and protein analysis of stem markers.
- Assessed proliferation patterns using confocal microscopy and validated model sensitivity with ATRA treatment.
Main Results:
- The developed spheroid model (SCESM) showed significantly increased stem marker expression compared to adherent cells and MCTS models.
- Proliferation was localized to the spheroid periphery, indicating stem cell enrichment.
- ATRA treatment effectively reduced stem marker expression, confirming model responsiveness.
Conclusions:
- The SCESM is a simple, HTS-compatible model for identifying anti-CSC compounds.
- This model enhances the screening of potential therapeutics targeting cancer stem cells.
- The SCESM offers a promising platform for developing new treatments to prevent tumor recurrence.
