Identification of serum biomarkers in HNSCC - U54 Pilot Project

Awarding Agency:  National Cancer Institute (NCI)
Award No:  2 U54 CA091408 06A1
Principal Investigator:  Dana Marshall, PhD

Grant Summary:
Early detection of pre-symptomatic tumors has proven to be clinically valuable in breast and cervical cancer resulting in reduction in the average tumor size at diagnosis and consequent improved patient survival. The development of biomarkers that can be used for early detection of head and neck squamous cell carcinoma (HNSCC) may similarly impact prognosis, since the majority of these tumors are identified at advanced stages. Additionally, HNSCC patients are at high risk for recurrence or second primary following initial therapy, and early detection of second primaries/recurrences is critical in order to offer the patient curative therapy. For both incident and recurrent HNSCC, a diagnostic blood test could improve early detection of these tumors and ultimately benefit patients. We have used mass spectrometry to identify putative serum biomarkers based on identification frequency. Our preliminary data, using multidimensional LC-MS/MS technology and accepted criteria to ensure correct protein identification, revealed that 600-800 proteins are routinely identified from 100ml of serum and that repeated analyses of the same samples further increases the number of solid identifications. The LC-MS/MS strategy we employed identified proteins of all molecular weights, as well as low abundance proteins. We have identified more than 10 candidate biomarkers based on differential identification between sera of cancer patients and controls. We propose to confirm that the proteins are differentially expressed in the identical serum from which they were identified using antibody and mass spectrometric quantitative techniques.  We will then analyze individual serum samples from cancer patients and controls to validate our findings. Sera samples will be collected at Meharry and Vanderbilt and cancer and control groups will be matched for race, age, sex, and current smoking history (self-reported and cotinine assay). Antibody-based analyses will be largely performed at Meharry, while mass spectrometric techniques will be performed at Vanderbilt.

This project will continue to solidify the collaboration between the head and neck group at Vanderbilt and the surgical research group at Meharry through shared resources, including diverse patient populations.   Additionally, this collaboration will support training of staff at MMC and support the development of an infrastructure for biomarker detection and confirmation and development of assays for this specific cancer, as well as others that are disproportionately represented in underserved populations.

Specific Aim I: Identify proteins whose serum levels differ between patients with and without HNSCC as measured by frequencies of peptide identification.  Proteins in serum pooled from: 1) 20 advanced stage HNSCC patients and; 2) 20 non-cancer controls will be identified. The difference in abundance of identified proteins between HNSCC patients and non-cancer controls will be determined based on the frequency of peptide identification.