Project description:Fusobacterium polymorphum has been implicated in oral squamous cell carcinoma, yet its ubiquitous presence in healthy individuals complicates its role as a disease driver and raises the possibility that some clinical isolates may possess greater pathogenic potential than others. To explore potential functional differences among isolates, we performed transcriptomic profiling of F. polymorphum strains obtained from across the oral epithelial disease spectrum. Despite being genetically indistinguishable, isolates derived from dysplastic lesions exhibited distinct transcriptional signatures, including significant upregulation of genes involved in heme uptake compared with isolates from healthy sites. These findings suggest that transcriptional heterogeneity among clinical isolates may contribute to functional differences that could influence their pathogenic potential.
Project description:Interspecies coaggregation promotes transcriptional changes of oral bacteria, contributing to the development of structurally balanced biofilms as well as oral diseases such as periodontitis. Streptococcus gordonii (S. gordonii) is an early colonizer of the oral cavity, and Fusobacterium nucleatum (F. nucleatum) may act as a bridge adhering to both early and late oral colonizers. These two species were commonly detected in healthy and periodontitis-diseased oral sites and could interact with immune cells such as macrophages. However, little research explored how intergeneric coaggregation affected transcriptional changes in S. gordonii and F. nucleatum subsp. polymorphum and how these gene changes might affect both species’ pathogenicity. The present study investigated transcriptional changes of both species in response to dual-species physical association using dual RNA-seq. Results indicated that after 30-min dual-species coaggregation, 148 genes were significantly up-regulated, and 124 genes were significantly down-regulated in S. gordonii. A total of 154 genes were significantly down-regulated, and 10 genes were significantly up-regulated in F. nucleatum subsp. polymorphum. A majority of up-regulated S. gordonii genes were involved in the biosynthesis and export of cell-wall proteins and the pathway of carbohydrate metabolism, and a group of down-regulated S. gordonii genes were associated with fatty acid biosynthesis and peptidoglycan biosynthesis. The transcriptome profiles indicated that the interspecies coaggregation led to a reduced level of DNA repair and lipopolysaccharides virulence in F. nucleatum subsp. polymorphum. The present study revealed that dual-species coaggregation induced a wide array of gene changes in S. gordonii and F. nucleatum subsp. polymorphum, enhancing S. gordonii’s adherence ability and attenuating F. nucleatum subsp. polymorphum's ability to produce LPS.
Project description:Fusobacterium nucleatum is a prominent member of the oral microbiota and is a common cause of human infection. F. nucleatum includes five subspecies: polymorphum, nucleatum, vincentii, fusiforme, and animalis. F. nucleatum subsp. polymorphum ATCC 10953 has been well characterized phenotypically and, in contrast to previously sequenced strains, is amenable to gene transfer. We sequenced and annotated the 2,429,698 bp genome of F. nucleatum subsp. polymorphum ATCC 10953. Plasmid pFN3 from the strain was also sequenced and analyzed. When compared to the other two available fusobacterial genomes (F. nucleatum subsp. nucleatum, and F. nucleatum subsp. vincentii) 627 open reading frames unique to F. nucleatum subsp. polymorphum ATCC 10953 were identified. A large percentage of these mapped within one of 28 regions or islands containing five or more genes. Seventeen percent of the clustered proteins that demonstrated similarity were most similar to proteins from the clostridia, with others being most similar to proteins from other gram-positive organisms such as Bacillus and Streptococcus. A ten kilobase region homologous to the Salmonella typhimurium propanediol utilization locus was identified, as was a prophage and integrated conjugal plasmid. The genome contains five composite ribozyme/transposons, similar to the CdISt IStrons described in Clostridium difficile. IStrons are not present in the other fusobacterial genomes. These findings indicate that F. nucleatum subsp. polymorphum is proficient at horizontal gene transfer and that exchange with the Firmicutes, particularly the Clostridia, is common.
Project description:Fusobacterium polymorphum has been implicated in oral squamous cell carcinoma, yet its widespread presence in the oral cavity raises questions about whether different clinical isolates exert distinct effects on host cells. To explore host transcriptional responses, we performed transcriptomic profiling of dysplastic oral keratinocytes (DOK) following exposure to F. polymorphum isolates derived from across the oral epithelial disease spectrum. The host transcriptomic response revealed a broadly shared pro-inflammatory signature across all isolates, regardless of their clinical origin. However, expression of genes and pathways associated with cell proliferation and related signaling processes varied among isolates and correlated with the dysplasia severity of the lesions from which the strains were derived. These findings suggest that while F. polymorphum isolates elicit a common inflammatory response in epithelial cells, variation in host transcriptional programs linked to proliferation may reflect differences in their biological impact.
Project description:Adhesin-mediated bacterial interspecies interactions are important elements in oral biofilm formation. They often occur on a species-specific level, which could determine health or disease association of a biofilm community. Among the key players involved in these processes are the ubiquitous fusobacteria that have been recognized for their ability to interact with numerous different binding partners. Fusobacterial interactions with Streptococcus mutans, an important oral cariogenic pathogen, have previously been described but most studies focused on binding to non-mutans streptococci and specific cognate adhesin pairs remain to be identified. Here, we demonstrated differential binding of oral fusobacteria to S. mutans. Screening of existing mutant derivatives indicated SpaP as the major S. mutans adhesin specific for binding to Fusobacterium nucleatum ssp. polymorphum but none of the other oral fusobacteria tested. We inactivated RadD, a known adhesin of F. nucleatum ssp. nucleatum for interaction with a number of gram-positive species, in F. nucleatum ssp. polymorphum and used a Lactococcus lactis heterologous SpaP expression system to demonstrate SpaP interaction with RadD of F. nucleatum ssp. polymorphum. This is a novel function for SpaP, which has mainly been characterized as an adhesin for binding to host proteins including salivary glycoproteins. In conclusion, we describe an additional role for SpaP as adhesin in interspecies adherence with RadD-SpaP as the interacting adhesin pair for binding between S. mutans and F. nucleatum ssp. polymorphum. Furthermore, S. mutans attachment to oral fusobacteria appears to involve species- and subspecies-dependent adhesin interactions.