Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma

BackgroundOral potentially malignant disorders (OPMDs), including conventional leukoplakia (OL) and proliferative verrucous leukoplakia (PVL), have distinct risks of progression to oral squamous cell carcinoma (OSCC). A role of the oral microbiome in this transformation is increasingly recognized, b...

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Main Authors: Rossella Intini, Sol Balsells, Leticia Bagan, Giulio Fortuna, Herve Sroussi, Jose Bagan
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Oral Health
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Online Access:https://www.frontiersin.org/articles/10.3389/froh.2025.1600090/full
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author Rossella Intini
Sol Balsells
Leticia Bagan
Giulio Fortuna
Herve Sroussi
Jose Bagan
Jose Bagan
author_facet Rossella Intini
Sol Balsells
Leticia Bagan
Giulio Fortuna
Herve Sroussi
Jose Bagan
Jose Bagan
author_sort Rossella Intini
collection DOAJ
description BackgroundOral potentially malignant disorders (OPMDs), including conventional leukoplakia (OL) and proliferative verrucous leukoplakia (PVL), have distinct risks of progression to oral squamous cell carcinoma (OSCC). A role of the oral microbiome in this transformation is increasingly recognized, but its contribution remains unclear.ObjectiveThis study aimed to analyze and compare the oral microbiota in patients with OL, PVL, and OSCC using 16S rRNA gene sequencing of saliva samples to identify microbial signatures associated with disease progression and to uncover potential biomarkers that would justify an aggressive treatment of OPMDs.MethodsSixty-six subjects with OPMDs were enrolled, comprising OL (n = 10), PVL (n = 28), and OSCC (n = 28). Saliva samples were collected, and DNA was extracted. The V3–V4 regions of the 16S rRNA gene were sequenced using the Illumina MiSeq platform. Bioinformatic analyses, including diversity assessments and taxonomic classification with the SILVA v138 database, were performed using QIIME2. Alpha diversity was evaluated with Chao1, Shannon, and Simpson indices, while beta diversity was assessed using Bray-Curtis and Jaccard distances.ResultsPVL exhibited the highest species richness, followed by OL, with OSCC showing the lowest diversity. While alpha diversity differences among the groups were not statistically significant (p > 0.05), beta diversity revealed distinct microbial community structures between OL and both PVL and OSCC (p < 0.05), but not between PVL and OSCC. At the phylum level, Firmicutes predominated across all groups, with significantly higher Actinobacteriota levels in OL (p = 0.002).ConclusionDistinct microbial patterns differentiate OL from PVL and OSCC, with OL being different from PVL and OSCC, suggesting progressive microbial dysbiosis in malignant transformation. These findings support the potential of oral microbiome profiling as a non-invasive diagnostic and prognostic tool in oral oncology and highlight the need for longitudinal studies to establish causal relationships.
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spelling doaj-art-b3acdf4973b4409ea33a85d7803b811e2025-08-20T03:09:48ZengFrontiers Media S.A.Frontiers in Oral Health2673-48422025-05-01610.3389/froh.2025.16000901600090Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinomaRossella Intini0Sol Balsells1Leticia Bagan2Giulio Fortuna3Herve Sroussi4Jose Bagan5Jose Bagan6Oral Medicine Unit, Stomatology Department, Valencia University, Valencia, SpainStatistical Advising Service, Fundació de Recerca Sant Joan de Déu, Esplugues de Llobregat, Spain.Oral Medicine Unit, Stomatology Department, Valencia University, Valencia, SpainDepartment of Medicine, Surgery and Dentistry, University of Salerno, Salerno, ItalyDivision of Oral Medicine and Dentistry, Brigham and Women’s Hospital and Dana Farber Cancer Institute Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, MA, United StatesMedicina Bucal Unit, Stomatology Department, Valencia University, Valencia, SpainPrecancer and Oral Cancer Research Group, Valencia University, Valencia, SpainBackgroundOral potentially malignant disorders (OPMDs), including conventional leukoplakia (OL) and proliferative verrucous leukoplakia (PVL), have distinct risks of progression to oral squamous cell carcinoma (OSCC). A role of the oral microbiome in this transformation is increasingly recognized, but its contribution remains unclear.ObjectiveThis study aimed to analyze and compare the oral microbiota in patients with OL, PVL, and OSCC using 16S rRNA gene sequencing of saliva samples to identify microbial signatures associated with disease progression and to uncover potential biomarkers that would justify an aggressive treatment of OPMDs.MethodsSixty-six subjects with OPMDs were enrolled, comprising OL (n = 10), PVL (n = 28), and OSCC (n = 28). Saliva samples were collected, and DNA was extracted. The V3–V4 regions of the 16S rRNA gene were sequenced using the Illumina MiSeq platform. Bioinformatic analyses, including diversity assessments and taxonomic classification with the SILVA v138 database, were performed using QIIME2. Alpha diversity was evaluated with Chao1, Shannon, and Simpson indices, while beta diversity was assessed using Bray-Curtis and Jaccard distances.ResultsPVL exhibited the highest species richness, followed by OL, with OSCC showing the lowest diversity. While alpha diversity differences among the groups were not statistically significant (p > 0.05), beta diversity revealed distinct microbial community structures between OL and both PVL and OSCC (p < 0.05), but not between PVL and OSCC. At the phylum level, Firmicutes predominated across all groups, with significantly higher Actinobacteriota levels in OL (p = 0.002).ConclusionDistinct microbial patterns differentiate OL from PVL and OSCC, with OL being different from PVL and OSCC, suggesting progressive microbial dysbiosis in malignant transformation. These findings support the potential of oral microbiome profiling as a non-invasive diagnostic and prognostic tool in oral oncology and highlight the need for longitudinal studies to establish causal relationships.https://www.frontiersin.org/articles/10.3389/froh.2025.1600090/fulloral microbiomeoral leukoplakiaproliferative verrucous leukoplakiaoral squamous cell carcinoma16S rRNA sequencingmicrobial diversity
spellingShingle Rossella Intini
Sol Balsells
Leticia Bagan
Giulio Fortuna
Herve Sroussi
Jose Bagan
Jose Bagan
Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma
Frontiers in Oral Health
oral microbiome
oral leukoplakia
proliferative verrucous leukoplakia
oral squamous cell carcinoma
16S rRNA sequencing
microbial diversity
title Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma
title_full Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma
title_fullStr Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma
title_full_unstemmed Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma
title_short Comparative analysis of oral microbiome in saliva samples of oral leukoplakia, proliferative leukoplakia and oral squamous cell carcinoma
title_sort comparative analysis of oral microbiome in saliva samples of oral leukoplakia proliferative leukoplakia and oral squamous cell carcinoma
topic oral microbiome
oral leukoplakia
proliferative verrucous leukoplakia
oral squamous cell carcinoma
16S rRNA sequencing
microbial diversity
url https://www.frontiersin.org/articles/10.3389/froh.2025.1600090/full
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