Metagenomic Analysis of the Oral Microbiome in Patients with Head and Neck Squamous Cell Carcinoma: A Case-Control Study
- Authors
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Niyati V. Thakkar
Faculty of Allied and Healthcare, Gokul Global University, Sidhpur, Gujarat, India -
Anisha Chaudhary
Quantum University Research Center, Quantum University, India -
Smiral Gameti
Faculty of Nursing, Parul University, Vadodara, Gujarat, India -
K. Ezhil Vendhan
Department of Ophthalmology, Vinayaka Mission’s Kirupananda Variyar Medical College & Hospitals, Vinayaka Mission’s Research Foundation (DU), Salem, Tamil Nadu, India -
Nittin Sharma
Center for Research Impact & Outcome, Chitkara University Institute of Engineering and Technology, Chitkara University, Rajpura, 140401, Punjab, India -
K.V. Jamuna
Department of Forensic Science, JAIN (Deemed-to-be University), Bengaluru, Karnataka, India -
Rajesh Kumar Lenka
Department of Microbiology, IMS and SUM Hospital, Siksha 'O' Anusandhan (Deemed to be University), Bhubaneswar, Odisha, India
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- Keywords:
- Head and Neck Squamous Cell Carcinoma (HNSCC), Oral Microbiome, Metagenomic Sequencing, Microbial Dysbiosis, Cancer Biomarkers, Oral Carcinogenesis, Microbial Biomarkers
- Abstract
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Head and neck squamous cell carcinoma (HNSCC) is one of the important cancers that are responsible for considerable morbidity all over the world. Recent studies reveal that changes in the oral microbiome are linked with HNSCC, but the exact microbial signatures according to the presence of disease need further research. In this study, metagenomic analysis was done on the oral microbiome of 210 HNSCC patients and 180 healthy controls based on the whole-genome shotgun metagenomic sequencing technique. Microbial diversity analysis, taxonomic profile, functional pathway analysis, and classification using a machine learning algorithm were performed for the identification of HNSCC-specific microbial signatures. HNSCC patients showed considerable changes in oral microbial profile as compared to the healthy subjects. The Shannon diversity index was significantly lower in patients with HNSCC than in control subjects (3.09 ± 0.34 vs 3.82 ± 0.41). An increase in the relative abundance of Fusobacterium, Prevotella, and Porphyromonas was observed in patients with HNSCC, while the relative abundance of the commensal species Streptococcus and Rothia was found to be decreased. The functional annotation revealed enrichment of pathways related to inflammation in the HNSCC samples. Using a random forest approach based on the microbial abundance profile, discrimination was possible between the two groups with a sensitivity of 82%, specificity of 79%, and AUC of 0.86 through cross-validation.
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[1] Su Mun L, Wye Lum S, Kong Yuiin Sze G, Hock Yoong C, Ching Yung K, Kah Lok L, Gopinath D. Association of microbiome with oral squamous cell carcinoma: a systematic review of the metagenomic studies. International Journal of Environmental Research and Public Health 2021; 18(14): 7224.
[2] Alzahem N, Al-Garadi MA, Chinnathambi A, Salmen SH, Wainwright M, Alharbi SA. Symphony of the Seas: Unveiling Metagenomic Biodiversity Through 16s RRNA in the Red Sea. Archives for Technical Sciences 2026; 1(35): 13-26.
[3] Ganly I, Hao Y, Rosenthal M, Wang H, Migliacci J, Huang B, Yang L. Oral microbiome in nonsmoker patients with oral cavity squamous cell carcinoma, defined by metagenomic shotgun sequencing. Cancers 2022; 14(24): 6096.
[4] Tusha, Tang Y, Ambhore V, Lavanya P, Thakur S. Gut Microbiome Modulation Through Personalized Nutrition for Metabolic Syndrome Management. International Academic Journal of Innovative Research 2025; 12(3): 79-85.
[5] Kwak S, Wang C, Usyk M, Wu F, Freedman ND, Huang WY, Hayes RB. Oral microbiome and subsequent risk of head and neck squamous cell cancer. JAMA Oncology 2024; 10(11): 1537-1547.
[6] Zhi J, Liang Y, Zhao W, Qiao J, Zheng Y, Peng X, Wang W. Oral microbiome-derived biomarkers for non-invasive diagnosis of head and neck squamous cell carcinoma. npj Biofilms and Microbiomes 2025; 11(1): 74.
[7] Ganly I, Pei Z, Hao Y, Ma Y, Rosenthal M, Wu Z, Yang L. Case control study comparing the HPV genome in patients with oral cavity squamous cell carcinoma to normal patients using metagenomic shotgun sequencing. Scientific Reports 2021; 11(1): 3867.
[8] Liu Y, Li Z, Qi Y, Wen X, Zhang L. Metagenomic analysis reveals a changing microbiome associated with the depth of invasion of oral squamous cell carcinoma. Frontiers in Microbiology 2022; 13: 795777.
[9] Aparna KG, Ravindra J, Chakraborty G, Ballamoole KK, Vinaya Kumar JR, Chakraborty A. 16S rRNA based metagenomic analysis unveils unique oral microbial signatures in oral squamous cell carcinoma cases from Coastal Karnataka, India. Acta Microbiologica et Immunologica Hungarica 2024; 71(3): 253-262.
[10] Torozan DA, Laczny CC, Roomp K, Wilmes P, Fleckenstein J, Schneider JG. Metagenomic Profiling of Oral Microbiome Dynamics During Chemoradiotherapy in Head and Neck Squamous Cell Carcinoma Patients. Cancer Medicine 2025; 14(1): e70589.
[11] Chan JY, Cheung MK, Lan L, Ng C, Lau EH, Yeung ZW, Chen Z. Characterization of oral microbiota in HPV and non-HPV head and neck squamous cell carcinoma and its association with patient outcomes. Oral Oncology 2022; 135: 106245.
[12] Srivastava A, Mishra S, Garg PK, Dubey AK, Deo SVS, Verma D. Comparative and analytical characterization of the oral bacteriome of smokeless tobacco users with oral squamous cell carcinoma. Applied Microbiology and Biotechnology 2022; 106(11): 4115-4128.
[13] Kim YK, Kwon EJ, Yu Y, Kim J, Woo SY, Choi HS, Kim YH. Microbial and molecular differences according to the location of head and neck cancers. Cancer Cell International 2022; 22(1): 135.
[14] Zhang M, Zhao Y, Umar A, Zhang H, Yang L, Huang J, Yu Z. Comparative analysis of microbial composition and functional characteristics in dental plaque and saliva of oral cancer patients. BMC Oral Health 2024; 24(1): 411.
[15] Chan JY, Ng CW, Lan L, Fung S, Li JW, Cai L, Chen Z. Restoration of the oral microbiota after surgery for head and neck squamous cell carcinoma is associated with patient outcomes. Frontiers in Oncology 2021; 11: 737843.
[16] Ahmad S, Jayamanne D, Bergamin S, Lawless A, Guminski A, Lee A, Oh B. Oral microbiome as a biomarker and therapeutic target in head and neck cancer: current insights and future directions. Cancers 2025; 17(16): 2667.
[17] Xie C, Yan Q, Zhang P, Zhang L, Yan G. Intratumoral microbiota omics analysis in head and neck squamous cell carcinoma. Frontiers in Microbiology 2025; 16: 1711139.
[18] Intini R, Balsells S, Bagan L, Fortuna G, Sroussi H, Bagan J. Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma. Frontiers in Oral Health 2025; 6: 1600090.
[19] Zhu H, Yip HC, Cheung MK, Chan HC, Ng C, Lau EH, Chen Z. Convergent dysbiosis of upper aerodigestive microbiota between patients with esophageal and oral cavity squamous cell carcinoma. International Journal of Cancer 2023; 152(9): 1903-1915.
[20] Lyu WN, Shen CY, Lee YH, Chen SK, Chuang EY, Lou PJ, Tsai MH. Novel biomarker identification for oral squamous cell carcinoma development in nonsmoker, nondrinker, and nonchewer patients using third-generation sequencing of oral microbiome. Journal of Oral Microbiology 2025; 17(1): 2565452.
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- 08-10-2026
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- Vol. 15 No. 4 (2026)
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