Current medical protocols for head and neck squamous cell carcinoma are deficient in viable biomarkers for more personalized therapeutic approaches. This study aims to uncover such potential biomarkers regarding the prediction of recurrence and delineating field cancerization in this type of cancer. Total RNA and microRNA next generation sequencing would grant us a comprehensive set of data that could guide the selection of such candidates for qRT-PCR validation and in situ hybridization topographic visualization.

Objectives:
1. Enlarging our biobank with samples from squamous cell carcinoma up to 120
patients with cancer of the oral cavity, oropharynx, hypopharynx and larynx. Sample collection would include tumor tissue, histologically normal peritumor mucosa, distant control mucosa and plasma.
2. Conduct total RNA and microRNA next-generation sequencing in a designated group of 10-12 patients from each subsite of cancer origin.
3. Conduct HPV infection analysis of the samples.
4. Conduct integrated analysis of the RNA profiles obtained from the next-generation sequencing and based on that, select candidate molecules with abnormal expression levels for possible biomarker potential and further investigation.

5. Conduct qRT-PCR validation of the selected RNA molecules across the whole cohort.
6. Analyze possible associations of these molecules with clinical characteristics and survival outcomes.
7. Detect RNA molecules with abnormal expression levels in histologically normal peritumor mucosa that have the same pattern of expression in tumor tissue, i.e., find potential biomarkers for field of cancerization.
8. Visualize some of the selected potential biomarkers with in situ hybridization.
9. Attempt to refine in situ hybridization techniques and methodology for accelerating the process of visualization with the help of phospholipid nanoparticles thus getting closer to a possible perioperative utilization of these findings and achieving translational application.