AUTHOR=Mougeot Jean-Luc C. , Beckman Micaela F. , Langdon Holden C. , Lalla Rajesh V. , Brennan Michael T. , Bahrani Mougeot Farah K. 

TITLE=Haemophilus pittmaniae and Leptotrichia spp. Constitute a Multi-Marker Signature in a Cohort of Human Papillomavirus-Positive Head and Neck Cancer Patients

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.794546

DOI=10.3389/fmicb.2021.794546

ISSN=1664-302X

ABSTRACT=<sec><title>Objectives</title><p>Human papillomavirus (HPV) is a known etiological factor of oropharyngeal head and neck cancer (HNC). HPV positivity and periodontal disease have been associated with higher HNC risk, suggesting a role for oral bacterial species. Our objective was to determine oral microbiome profiles in HNC patients (HPV-positive and HPV-negative) and in healthy controls (HC).</p></sec><sec><title>Methods</title><p>Saliva samples and swabs of buccal mucosa, supragingival plaque, and tongue were collected from HNC patients (<italic>N</italic> = 23 patients, <italic>n</italic> = 92 samples) before cancer therapy. Next-generation sequencing (16S-rRNA gene V3–V4 region) was used to determine bacterial taxa relative abundance (RA). β-Diversities of HNC HPV+ (<italic>N</italic> = 16 patients, <italic>n</italic> = 64 samples) and HNC HPV– (<italic>N</italic> = 7 patients, <italic>n</italic> = 28 samples) groups were compared using PERMANOVA (pMonte Carlo &lt; 0.05). LEfSe discriminant analysis was performed to identify differentiating taxa (Log LDA &gt; 2.0). RA differences were analyzed by Mann–Whitney <italic>U</italic>-test (α = 0.05). CombiROC program was used to determine multi-marker bacterial signatures. The Microbial Interaction Network Database (MIND) and LitSuggest online tools were used for complementary analyses.</p></sec><sec><title>Results</title><p>HNC vs. HC and HNC HPV+ vs. HNC HPV– β-diversities differed significantly (pMonte Carlo &lt; 0.05). <italic>Streptococcus</italic> was the most abundant genus for HNC and HC groups, while <italic>Rothia mucilaginosa</italic> and <italic>Haemophilus parainfluenzae</italic> were the most abundant species in HNC and HC patients, respectively, regardless of antibiotics treatment. LEfSe analysis identified 43 and 44 distinctive species for HNC HPV+ and HNC HPV– groups, respectively. In HNC HPV+ group, 26 periodontal disease-associated species identified by LefSe had a higher average RA compared to HNC HPV– group. The significant species included <italic>Alloprevotella tannerae</italic>, <italic>Fusobacterium periodonticum</italic>, <italic>Haemophilus pittmaniae</italic>, <italic>Lachnoanaerobaulum orale</italic>, and <italic>Leptotrichia</italic> spp. (Mann–Whitney <italic>U</italic>-test, <italic>p</italic> &lt; 0.05). Of 43 LEfSe-identified species in HPV+ group, 31 had a higher RA compared to HPV– group (Mann–Whitney <italic>U</italic>-test, <italic>p</italic> &lt; 0.05). MIND analysis confirmed interactions between <italic>Haemophilus</italic> and <italic>Leptotrichia</italic> spp., representing a multi-marker signature per CombiROC analysis [area under the curve (AUC) &gt; 0.9]. LitSuggest correctly classified 15 articles relevant to oral microbiome and HPV status.</p></sec><sec><title>Conclusion</title><p>Oral microbiome profiles of HNC HPV+ and HNC HPV– patients differed significantly regarding periodontal-associated species. Our results suggest that oral bacterial species (e.g., <italic>Leptotrichia</italic> spp.), possessing unique niches and invasive properties, coexist with HPV within HPV-induced oral lesions in HNC patients. Further investigation into host–microbe interactions in HPV-positive HNC patients may shed light into cancer development.</p></sec>