Deciphering the Network Architecture of APOBEC3-Driven Mutagenesis in HPV-Positive Head and Neck Cancers.

UNLABELLED: APOBEC3A (A3A) and APOBEC3B (A3B) are cytosine deaminases that restrict viral infection and can also mutate the host genome. In human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC), expression of both enzymes is elevated, but bulk sequencing averages their effects across mixed cell populations. Here we profile single cells from HPV16-positive HNSCC tumors and matched normal tissue. We find that the APOBEC3 (A3) single base substitution mutational signature, SBS2, is enriched in cells expressing more A3A than A3B , whereas copy number alteration (CNA) burden is enriched in cells expressing more A3B than A3A . Tumor versus normal co-expression networks identify the A3 interactors RALY and HNRNPA2B1 as candidate A3 activators. We find that SBS2 and CNA mark the maintenance and virus assembly stages of the HPV16 life cycle, and their ratio may offer a molecular estimate of tumor age. The neoantigens from immune-visible SBS2-HIGH and immune-evasive CNA-HIGH cells identify candidates for mRNA vaccines matched to a tumor's viral state.

SIGNIFICANCE: 3Single-cell analysis of HPV-positive HNSCC reveals that DNA editing enzymes A3A and A3B associate with distinct HPV16 life cycle stages and immunological tumor states, enabling stage-matched neoantigen vaccine candidate identification for personalized immunotherapy.