Curative treatment of localized or locally advanced head and neck squamous cell carcinomas (HNSCC) necessitates precise diagnosis, surgery, and drug therapy. Minimal residual disease (MRD) diagnostics plays a promising role to identify patients who will benefit from an additional adjuvant drug treatment after tumor resection with curative intent.

The analysis of circulating cell-free tumor DNA (ctDNA) in blood presents a rapidly evolving field in oncology, allowing for primary diagnostics, tumor profiling, screening, early therapy response and resistance monitoring, therapy selection, local recurrence diagnostics and MRD detection. Both tumor-informed and tumor-agnostic approaches have demonstrated the ability to detect few or multiple cancer-specific mutations and human papillomavirus (HPV) sequences, which have already shown high clinical performance of MRD testing in several tumor entities. Recently, the effectiveness of DNA methylation biomarkers in detecting MRD has already been demonstrated. However, multimodal MRD tests that combine mutation, HPV, and DNA methylation testing, hold great potential to improve MRD testing but have until now not been clinically validated. The present project aims to develop and assess the clinical performance of a tumor-agnostic next-generation sequencing (NGS) MRD test that combines TP53 mutation, HPV18 and 16, and aberrant DNA methylation testing in a single tube reaction. Our proof-of-concept and clinical performance evaluation study will exemplarily analyse a prospective cohort of 200 patients with localized or locally advanced HNSCC.

Nein