Functional signatures of oral dysbiosis during periodontitis progression revealed by microbial metatranscriptome analysis.
BACKGROUND
Periodontitis is a polymicrobial biofilm-induced inflammatory disease that affects 743 million people worldwide. The current model to explain periodontitis progression proposes that changes in the relative abundance of members of the oral microbiome lead to dysbiosis in the host-microbiome crosstalk and then to inflammation and bone loss. Using combined metagenome/metatranscriptome analysis of the subgingival microbiome in progressing and non-progressing sites, we have characterized the distinct molecular signatures of periodontitis progression.


METHODS
Metatranscriptome analysis was conducted on samples from subgingival biofilms from progressing and stable sites from periodontitis patients. Community-wide expression profiles were obtained using Next Generation Sequencing (Illumina). Sequences were aligned using 'bowtie2' against a constructed oral microbiome database. Differential expression analysis was performed using the non-parametric algorithm implemented on the R package 'NOISeqBio'. We summarized global functional activities of the oral microbial community by set enrichment analysis based on the Gene Ontology (GO) orthology.


RESULTS
Gene ontology enrichment analysis showed an over-representation in the baseline of active sites of terms related to cell motility, lipid A and peptidoglycan biosynthesis, and transport of iron, potassium, and amino acids. Periodontal pathogens (Tannerella forsythia and Porphyromonas gingivalis) upregulated different TonB-dependent receptors, peptidases, proteases, aerotolerance genes, iron transport genes, hemolysins, and CRISPR-associated genes. Surprisingly, organisms that have not been usually associated with the disease (Streptococcus oralis, Streptococcus mutans, Streptococcus intermedius, Streptococcus mitis, Veillonella parvula, and Pseudomonas fluorenscens) were highly active transcribing putative virulence factors. We detected patterns of activities associated with progression of clinical traits. Among those we found that the profiles of expression of cobalamin biosynthesis, proteolysis, and potassium transport were associated with the evolution towards disease.


CONCLUSIONS
We identified metabolic changes in the microbial community associated with the initial stages of dysbiosis. Regardless of the overall composition of the community, certain metabolic signatures are consistent with disease progression. Our results suggest that the whole community, and not just a handful of oral pathogens, is responsible for an increase in virulence that leads to progression.


BACKGROUND
NCT01489839, 6 December 2011.
Forsyth Institute, 245 First Street, Cambridge, Massachusetts 02142 USA.
Susan
S
Yost
Forsyth Institute, 245 First Street, Cambridge, Massachusetts 02142 USA.
Ana E
AE
Duran-Pinedo
University of North Carolina Chapel Hill, School of Dentistry, Chapel Hill, NC 27599-7450 USA.
Ricardo
R
Teles
Forsyth Institute, 245 First Street, Cambridge, Massachusetts 02142 USA.
Keerthana
K
Krishnan
Forsyth Institute, 245 First Street, Cambridge, Massachusetts 02142 USA ; Harvard School of Dental Medicine, 188 Longwood Ave, Boston, MA 02115 USA.
Jorge
J
Frias-Lopez