Clonal relationship and alcohol consumption-associated mutational signature in synchronous hypopharyngeal tumours and oesophageal squamous cell carcinoma

Abstract

Background The patients with dual oesophageal squamous cell carcinoma (ESCC) and hypopharyngeal cancer (HPC) have poor prognosis; their underlying genetic pathogenesis is unclear. We hypothesise that development of synchronous ESCC/HPC depends on multicentricity or independent origin, rather than multifocality due to local or lateral spreading. Method Multiple region whole-exome sequencing (M-WES) and clonality analysis were used to assess clonal relationship and spatial inter- or intra-tumour heterogeneity (ITH) in 62 tumour regions from eight dual ESCC/HPC and ten ESCC patients. Results All synchronous ESCC/HPC patients had COSMIC 16 mutation signatures, compared to only 40% ESCC in the current study (p = 0.013) and public data set (n = 165, p = 0.003). This alcohol consumption-related mutation signature 16, commonly involved in multiple alcohol-related cancers, was significantly associated with drinking and alcohol metabolism-related ADH1B rs1229984. The mutational landscape and copy number profiles were completely distinct between the two primary tumours; clonality analysis further suggested the two primary tumours shared no or only one clone accompanying independent subclone evolution. M-WES strategy demonstrated higher sensitivity and accuracy for detection of mutational prevalence and the late branch mutations among different regions in the ESCC tumours, compared to traditional sequencing analysis based on single biopsy strategy. Patients with high ITH assessed by cancer cell fraction analysis after M-WES were significantly associated with both relapse and survival. Conclusions Our hypothesis-generating M-WES ITH assessment data have implications for prognostication. Collectively, our findings support multicentric independent clonal evolution, the field cancerisation theory, and suggest novel insights implicating an aetiologic role of alcohol metabolism in dual ESCC/HPC carcinogenesis.

Circulating Tumor DNA Dynamics as Prognostic Markers in Locally Advanced and Metastatic Esophageal Squamous Cell Carcinoma

“…Formalin-fixed paraffin-embedded (FFPE) tissues were processed and analyzed using the AVENIO tumor tissue expanded kit (F. Hoffmann-La Roche Ltd), according to the manufacturer’s instructions. Briefly, DNA was isolated from FFPE sections after macrodissection for library preparation and sequenced as previously described . Data were analyzed using AVENIO oncology analysis software, version 1.2 with default setting.…”

“…Data were analyzed using AVENIO oncology analysis software, version 1.2 with default setting. Genomic DNA extraction, library preparation, and data analysis from snap-frozen tissues were performed as previously described …”

“…Briefly, DNA was isolated from FFPE sections after macrodissection for library preparation and sequenced as previously described. 11 Data were analyzed using AVENIO oncology analysis software, version 1.2 with default setting. Genomic DNA extraction, library preparation, and data analysis from snap-frozen tissues were performed as previously described.…”

“…Genomic DNA extraction, library preparation, and data analysis from snap-frozen tissues were performed as previously described. 11…”

Successful surgery with preservation of laryngeal function in a patient with collision primary squamous cell carcinoma and adenoid cystic carcinoma in the hypopharynx and synchronous esophageal carcinoma: A case report

“…Whether the prognosis of hypopharyngeal collision SCC is poorer than that of isolated hypopharyngeal ACC remains unclear, but a hypopharyngeal SCC combined with an esophageal carcinoma has a poorer prognosis than hypopharyngeal carcinoma alone. [31][32][33] Hypopharyngeal carcinoma is often combined with a later esophageal primary cancer, contributing to the poor prognosis. 34 Studies have also shown that the second primary esophageal carcinoma is one of the important causes of treatment failure and death in patients with hypopharyngeal cancer.…”