Abstract
Minimally invasive techniques are required for the identification of head and neck cancer (HNC) patients who are at an increased risk of metastasis, or are not responding to therapy. An approach utilised in other solid cancers is the identification and enumeration of circulating tumour cells (CTCs) in the peripheral blood of patients. Low numbers of CTCs has been a limiting factor in the HNC field to date. Here we present a methodology to expand HNC patient derived CTCs ex-vivo. As a proof of principle study, 25 advanced stage HNC patient bloods were enriched for circulating tumour cells through negative selection and cultured in 2D and 3D culture environments under hypoxic conditions (2% O2, 5% CO2). CTCs were detected in 14/25 (56%) of patients (ranging from 1–15 CTCs/5 mL blood). Short term CTC cultures were successfully generated in 7/25 advanced stage HNC patients (5/7 of these cultures were from HPV+ patients). Blood samples from which CTC culture was successful had higher CTC counts (p = 0.0002), and were predominantly from HPV+ patients (p = 0.007). This is, to our knowledge, the first pilot study to culture HNC CTCs ex-vivo. Further studies are warranted to determine the use of short term expansion in HNC and the role of HPV in promoting culture success.
Highlights
Head and neck cancers are the seventh most common cancer globally
Circulating tumour cells (CTCs) found in the lymphovasculature are thought to have the propensity to metastasize at distant sites
A total of 25 head and neck cancer (HNC) patients were recruited for this study and for all samples circulating tumour cells (CTCs) cultures attempted
Summary
Head and neck cancers are the seventh most common cancer globally. Approximately 50% of the patients die within 5 years. Current diagnostic methods include clinical assessment, imaging and tissue biopsy [1]. When metastases are clinically evident, treatment is palliative in nature only. There are currently no methods to predict which patients with a higher disease burden will develop metastases. Circulating tumour cells (CTCs) found in the lymphovasculature are thought to have the propensity to metastasize at distant sites. Enrichment strategies aim to increase the CTC concentration from blood by several log units for easier detection by downstream methodologies. Biological approaches for CTC enrichment can include positive selection using anti-epithelial marker www.impactjournals.com/oncotarget antibodies (EpCAM), anti-mesenchymal marker antibodies (N-cadherin) or negative selection using antibodies against CD45 to deplete leukocytes [3]. The physical properties of CTCs can be employed for cell separation
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