Intratumoral microbial networks as biomarkers for second primary oral cancer risk in esophageal squamous cell carcinoma

Wei-Ni Lyu; Cheng-Ying Shen; Yi-Jian Tsai; Li-Han Chen; Yung-Hua Lee; Shin-Kuang Chen; Jang-Ming Lee; Pei-Jen Lou; Eric Y Chuang; Mong-Hsun Tsai

doi:10.1038/s41598-026-52247-7

Intratumoral microbial networks as biomarkers for second primary oral cancer risk in esophageal squamous cell carcinoma

Sci Rep. 2026 May 8. doi: 10.1038/s41598-026-52247-7. Online ahead of print.

Authors

Wei-Ni Lyu^#^{1

2}, Cheng-Ying Shen^#¹, Yi-Jian Tsai^#^{3

4}, Li-Han Chen⁵, Yung-Hua Lee¹, Shin-Kuang Chen⁶, Jang-Ming Lee⁷, Pei-Jen Lou⁸, Eric Y Chuang⁹, Mong-Hsun Tsai¹⁰

Affiliations

¹ Institute of Biotechnology, National Taiwan University, No. 81, Changxing St., Taipei, 106, Taiwan.
² Health Innovation Center, National Yang Ming Chiao Tung University, Taipei, Taiwan.
³ Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, 106, Taiwan.
⁴ Division of Colorectal Surgery, Department of Surgery, Fu Jen Catholic University Hospital, New Taipei, Taiwan.
⁵ Institute of Fisheries Science, National Taiwan University, Taipei, Taiwan.
⁶ Center for Biotechnology, National Taiwan University, Taipei, Taiwan.
⁷ Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan.
⁸ Department of Otolaryngology, National Taiwan University Hospital, Taipei, Taiwan.
⁹ Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, 106, Taiwan. chuangey@ntu.edu.tw.
¹⁰ Institute of Biotechnology, National Taiwan University, No. 81, Changxing St., Taipei, 106, Taiwan. motiont@ntu.edu.tw.

^# Contributed equally.

PMID: 42103910
DOI: 10.1038/s41598-026-52247-7

Abstract

Esophageal squamous cell carcinoma (ESCC) survivors remain at elevated risk of developing second primary oral cancer (SPOC), yet the role of intratumoral microbiomes in SPOC emergence is not fully understood. We performed 16 S rRNA V3-V4 sequencing on tumor brushings from 28 ESCC patients (20 SPOC-negative, 8 SPOC-positive) to profile microbial diversity, taxonomic composition, functional potential, and interaction networks. Alpha diversity metrics (Chao1, Shannon) did not differ significantly between groups (p > 0.05), whereas sparse partial least squares-discriminant analysis of beta diversity robustly separated SPOC-positive from SPOC-negative tumors (p < 0.001), identifying 32 discriminant amplicon sequence variants (ASVs) linked to 41 differential KEGG pathways. Intratumoral Spearman correlation networks (|r| > 0.3, p < 0.05) between the ten most abundant genera and these pathways revealed two distinct modules: a SPOC-associated network centered on Prevotella pallens and P. scopos, enriched in carbohydrate metabolism, PI3K-Akt signaling, and glycosaminoglycan degradation; and a non-SPOC network anchored by Alcaligenaceae, Cyanobiaceae, Rhodobacteraceae, and Prevotella oris, associated with macrolide biosynthesis and aminobenzoate degradation. These findings demonstrate that specific intratumoral microbial interaction networks distinguish ESCC patients who develop SPOC, and highlight network-based microbial signatures as promising biomarkers for SPOC risk stratification.

Keywords: Esophageal squamous cell carcinoma; Microbial biomarkers; Microbiome; Second primary oral cancer.

Abstract

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