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Updated: Oct 17, 2025
Therapy Testing in a Spheroid-based 3D Cell Culture Model for Head and Neck Squamous Cell Carcinoma
Published on: April 20, 2018
Metabolic Plasticity and Combinatorial Radiosensitisation Strategies in Human Papillomavirus-Positive Squamous Cell
Mark D Wilkie1,2, Emad A Anaam1, Andrew S Lau1,2
1Cancer Research Centre, Department of Molecular & Clinical Cancer Medicine, The University of Liverpool, 200 London Road, Liverpool L3 9TA, UK.
Background:
A major objective in the management of human papillomavirus (HPV)-positive squamous cell carcinoma of the head and neck (SCCHN) is to reduce long-term functional ramifications while maintaining oncological outcomes. This study examined the metabolic profile of HPV-positive SCCHN and the potential role of anti-metabolic therapeutics to achieve radiosensitisation as a potential means to de-escalate radiation therapy.
Methods:
Three established HPV-positive SCCHN cell lines were studied (UM-SCC-104, UPCI:SCC154, and VU-SCC-147), together with a typical TP53 mutant HPV-negative SCCHN cell line (UM-SCC-81B) for comparison. Metabolic profiling was performed using extracellular flux analysis during specifically designed mitochondrial and glycolytic stress tests. Sensitivity to ionising radiation (IR) was evaluated using clonogenic assays following no treatment, or treatment with: 25 mM 2-deoxy-D-glucose (glycolytic inhibitor) alone; 20 mM metformin (electron transport chain inhibitor) alone; or 25 mM 2-deoxy-D-glucose and 20 mM metformin combined. Expression levels of p53 and reporters of p53 function (MDM2, p53, Phospho-p53 [Ser15], TIGAR and p21 [CDKN1A]) were examined by western blotting.
Results:
HPV-positive SCCHN cell lines exhibited a diverse metabolic phenotype, displaying robust mitochondrial and glycolytic reserve capacities. This metabolic profile, in turn, correlated with IR response following administration of anti-metabolic agents, in that both 2-deoxy-D-glucose and metformin were required to significantly potentiate the effects of IR in these cell lines.
Conclusions:
In contrast to our recently published data on HPV-negative SCCHN cells, which display relative glycolytic dependence, HPV-positive SCCHN cells can only be sensitised to IR using a complex anti-metabolic approach targeting both mitochondrial respiration and glycolysis, reflecting their metabolically diverse phenotype. Notionally, this may provide an attractive platform for treatment de-intensification in the clinical setting by facilitating IR dose reduction to minimise the impact of treatment on long-term function.
Insights
Human papillomavirus (HPV)-positive head and neck cancers require a dual anti-metabolic therapy targeting both glycolysis and mitochondrial respiration to enhance radiosensitization. This approach may allow for reduced radiation doses, minimizing long-term side effects.
Area of Science:
- Oncology
- Metabolic Research
- Radiation Therapy
Background:
- Human papillomavirus (HPV)-positive squamous cell carcinoma of the head and neck (SCCHN) management aims to balance oncological outcomes with reduced long-term functional deficits.
- Investigating the metabolic profile of HPV-positive SCCHN is crucial for developing targeted therapies.
- This study explores the potential of anti-metabolic drugs to sensitize tumors to radiation, potentially enabling treatment de-escalation.
Purpose of the Study:
- To characterize the metabolic phenotype of HPV-positive SCCHN cell lines.
- To evaluate the radiosensitizing effects of anti-metabolic agents targeting glycolysis and mitochondrial respiration.
- To determine if a combined anti-metabolic strategy can potentiate the effects of ionizing radiation (IR) in HPV-positive SCCHN.
Main Methods:
- Utilized three HPV-positive SCCHN cell lines and one HPV-negative cell line for comparative metabolic profiling via extracellular flux analysis.
- Assessed radiosensitivity using clonogenic assays after treatment with glycolytic inhibitor (2-deoxy-D-glucose) and/or mitochondrial inhibitor (metformin).
- Analyzed expression of p53 and related proteins (MDM2, TIGAR, p21) using western blotting.
Main Results:
- HPV-positive SCCHN cell lines demonstrated significant mitochondrial and glycolytic reserve capacities, indicating metabolic diversity.
- A combination of 2-deoxy-D-glucose and metformin was necessary to significantly enhance the radiosensitizing effects of ionizing radiation in these cell lines.
- The metabolic profile correlated with the response to combined anti-metabolic agents and IR.
Conclusions:
- Unlike HPV-negative SCCHN cells, HPV-positive SCCHN cells require a dual approach targeting both glycolysis and mitochondrial respiration for radiosensitization.
- The diverse metabolic phenotype of HPV-positive SCCHN necessitates a complex anti-metabolic strategy.
- This approach offers a potential strategy for de-intensifying radiation therapy, reducing treatment-related toxicity and preserving long-term function.
