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

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

doi:10.3389/fmicb.2021.794546

Jean-Luc C Mougeot, Farah K Bahrani Mougeot + Show 4 more

Open Access

https://doi.org/10.3389/fmicb.2021.794546

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Abstract

ObjectivesHuman 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).MethodsSaliva samples and swabs of buccal mucosa, supragingival plaque, and tongue were collected from HNC patients (N = 23 patients, n = 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+ (N = 16 patients, n = 64 samples) and HNC HPV– (N = 7 patients, n = 28 samples) groups were compared using PERMANOVA (pMonte Carlo < 0.05). LEfSe discriminant analysis was performed to identify differentiating taxa (Log LDA > 2.0). RA differences were analyzed by Mann–Whitney U-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.ResultsHNC vs. HC and HNC HPV+ vs. HNC HPV– β-diversities differed significantly (pMonte Carlo < 0.05). Streptococcus was the most abundant genus for HNC and HC groups, while Rothia mucilaginosa and Haemophilus parainfluenzae 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 Alloprevotella tannerae, Fusobacterium periodonticum, Haemophilus pittmaniae, Lachnoanaerobaulum orale, and Leptotrichia spp. (Mann–Whitney U-test, p < 0.05). Of 43 LEfSe-identified species in HPV+ group, 31 had a higher RA compared to HPV– group (Mann–Whitney U-test, p < 0.05). MIND analysis confirmed interactions between Haemophilus and Leptotrichia spp., representing a multi-marker signature per CombiROC analysis [area under the curve (AUC) > 0.9]. LitSuggest correctly classified 15 articles relevant to oral microbiome and HPV status.ConclusionOral microbiome profiles of HNC HPV+ and HNC HPV– patients differed significantly regarding periodontal-associated species. Our results suggest that oral bacterial species (e.g., Leptotrichia 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.

Highlights

Head and neck cancer (HNC) is the sixth most common cancer worldwide with over 95% comprising squamous cell carcinomas (SCCs) (Jemal et al, 2008; Kumarasamy et al, 2019)
While no significant differences were noted in age and ethnicity, the male population was over-represented in the HNC patient set in
Three were reviews and 12 were research articles evaluating the human papillomavirus (HPV) status in the context of oral tumor and microbiome relationship in SCC patients with oropharynx, including tonsil as the primary tumor site (Table 3). This is the first study to evaluate the microbial differences in HNC HPV+ patients compared to those of healthy individuals and HNC HPV– patients by means of oral samples including saliva, buccal mucosa, supragingival plaque, and tongue swabs using multivariate analysis

Summary

Introduction

Head and neck cancer (HNC) is the sixth most common cancer worldwide with over 95% comprising squamous cell carcinomas (SCCs) (Jemal et al, 2008; Kumarasamy et al, 2019). Head and neck SCCs are characterized by a locoregional development mainly diagnosed at an advanced stage of the disease, resulting in difficult treatment and eradication of both pre-neoplastic and neoplastic tissue (Carvalho et al, 2005; Ganci et al, 2015). Despite advancements in chemoradiation, ionizing radiation, and surgical resection techniques, HNC has an overall mortality rate of approximately 50% and is characterized by high recurrence rates (Carvalho et al, 2005). While triggers of HNC development have not been fully elucidated, two primary risk factors have been identified, namely, alcohol and tobacco consumptions (Božinovicet al., 2019). HPV-associated SCCs represent the most common HPV-related cancer in the US and are classified by a new staging system for oropharyngeal cancers (National Cancer Institute1: van Gysen et al, 2019)

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