# Liquid Biopsies in HNSCC: Current Landscape and Emerging Opportunities in the Era of HPV Stratification

## TL;DR

This paper reviews how liquid biopsies can improve HNSCC treatment by detecting cancer recurrence using blood-based markers, with a focus on HPV status differences.

## Contribution

The paper highlights the clinical readiness differences between ctDNA, exosomes, and CTCs in HPV-positive HNSCC and proposes a translational pathway for standardization and validation.

## Key findings

- HPV-related ctDNA is nearing clinical validation for MRD detection in HPV-positive HNSCC.
- Exosomes and CTCs face technical challenges like lack of standardized assays and limited reproducibility.
- Divergent analytical platforms are needed due to differences in HPV-positive and HPV-negative tumor biology.

## Abstract

Head and neck squamous cell carcinoma (HNSCC) is biologically and clinically dichotomous according to HPV status, a distinction that fundamentally dictates the design, implementation, and interpretation of liquid biopsy strategies. Conventional anatomical imaging lacks sufficient sensitivity for minimal residual disease (MRD) detection, contributing significantly to treatment failure and suboptimal clinical outcomes. This review provides a critical, evidence-based synthesis of the three principal circulating analytes, circulating tumor DNA (ctDNA), exosomes, and circulating tumor cells (CTCs), and their evolving roles in real-time, non-invasive molecular monitoring. Critically, the clinical readiness of these analytes differs substantially: while ctDNA, particularly HPV-related ctDNA, is approaching clinical validation for MRD detection and recurrence surveillance in HPV-positive HNSCC, exosomes and CTCs remain investigational tools hindered by ongoing technical challenges including lack of standardized assays, limited reproducibility across platforms, and insufficient prospective validation. We review how the presence of a clonal, virally derived DNA target in HPV-positive HNSCC contrasts with the heterogeneous somatic mutational landscape of HPV-negative tumors, necessitating divergent analytical platforms and yielding distinct clinical utility profiles for MRD detection and recurrence surveillance. We further outline a pragmatic translational pathway focused on assay standardization, particularly for exosomes and CTCs where this foundational work is most urgently needed, integration of complementary multimodal liquid biopsy approaches, and rigorously designed prospective interventional clinical trials to establish clinical utility. Collectively, these efforts aim to transition HNSCC management from reactive, anatomy-based surveillance to proactive, molecularly guided precision oncology, with the potential to improve therapeutic decision-making and patient outcomes.