Development of the First Tractable Genetic System for Parvimonas micra, a Ubiquitous Pathobiont in Human Dysbiotic Disease

Higashi, Dustin L.; McGuire, Sean E.; AbdelRahman, Yasser M.; Zou, Zhengzhong; Qin, Huashu; Anderson, David A.; Palmer, Elizabeth; Kreth, Jens; Merritt, Justin

doi:10.1128/spectrum.00465-22

Cited by 10 publications

(11 citation statements)

References 56 publications

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“…Periodontal disease has previously been described as having inflammophilic bacterial communities, meaning the species present thrive under inflammatory conditions, allowing them to outcompete other species 34, 35 . Parvimonas and Fusobacterium , as part of microbial cohorts, were shown to reduce neutrophil killing via dampening of reactive oxygen species production, and produced proteases to degrade immunoglobulins and complement proteins, reducing phagocytosis by host immune cells 35 . Both Parvimonas and Fusobacterium have been shown to induce inflammatory cytokines like IL-6 and IL-8 35, 36 .…”

Section: Discussionmentioning

confidence: 99%

“…Parvimonas and Fusobacterium , as part of microbial cohorts, were shown to reduce neutrophil killing via dampening of reactive oxygen species production, and produced proteases to degrade immunoglobulins and complement proteins, reducing phagocytosis by host immune cells 35 . Both Parvimonas and Fusobacterium have been shown to induce inflammatory cytokines like IL-6 and IL-8 35, 36 . Fusobacterium was shown to invade the epithelial barrier in appendicitis 25 .…”

Section: Discussionmentioning

confidence: 99%

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Germinal center BCR maturation in appendicitis reveals a role for antigen-specific adaptive immune responses during disease

Stewart,

Taghvaei,

Leon

et al. 2024

Preprint

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Appendicitis is one of the most common abdominal emergencies globally, yet little is understood about the inflammatory mechanisms or potential drivers of disease. Neutrophil inflammation and increased cytokine expression such as IL-6 and IL-8 are hallmarks of appendicitis inflammation. However, early histological studies identified increased T and B cell infiltration during appendicitis, providing support for adaptive immune activation as well, although this has never been investigated in depth. We hypothesized that antigen-dependent activation of the adaptive immune response contributes to appendicitis pathology, in addition to the known innate-mediated processes. Via a series of transcriptomic approaches and lymphocyte repertoire analysis in human appendiceal tissue, we identified evidence of antigen-dependent B cell activation. Increased somatic hypermutation in the germinal center and plasma cell compartment was comprised of presumed high-affinity IgG and IgA B cells. We propose that the appendiceal microbiome acts as a source of antigen, as significant microbial dysbiosis was observed during appendicitis. This dysbiosis was characterized by outgrowth of pathobionts such as Parvimonas and oral biofilm-formers such as Fretibacterium and Fusobacterium, in line with previous reports. We also identified potential loss of epithelial barrier integrity via spatial transcriptomic analysis of the appendiceal epithelium, supporting the possibility of microbial invasion into the tissue during appendicitis. This study provides insight into the inflammatory mechanisms of a common disease and helps to define the immune and microbial compartment of an often-ignored organ, the appendix.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Germinal center BCR maturation in appendicitis reveals a role for antigen-specific adaptive immune responses during disease

Stewart,

Taghvaei,

Leon

et al. 2024

Preprint

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“…We identified a highly diverse MecA interactome and were able to use this information to develop and validate a new split luciferase complementation assay (SLCA) useful for monitoring PPI in live bacteria. Our SLCA approach employed a synthetic red-shifted Renilla luciferase variant of RLuc8 (61–64) called Green Renilla (RenG), which we have previously demonstrated to yield exceptionally high activity in a variety of bacterial species (46,65). Besides its robust luminescence output, RenG is also particularly well suited for SLCA applications due to its small size (36 kDa) and monomeric functionality.…”

Section: Discussionmentioning

confidence: 99%

Mass spectrometry and split luciferase complementation assays reveal the MecA protein interactome ofStreptococcus mutans

Qin,

Anderson,

Zou

et al. 2023

Preprint

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MecA is a highly conserved adaptor protein encoded by prokaryotes from theBacillotaphylum. MecA mutants exhibit similar pleiotropic defects in a variety of organisms, although most of these phenotypes currently lack a mechanistic basis. MecA mediates ClpCP-dependent proteolysis of its substrates, but only several such substrates have been reported in the literature and there are suggestions that proteolysis-independent regulatory mechanisms may also exist. Here, we provide the first comprehensive characterization of the MecA interactome and further assess its regulatory role in Clp-dependent proteolysis. Untargeted coimmunoprecipitation assays coupled with mass spectrometry revealed that the MecA ortholog from the oral pathobiontStreptococcus mutanslikely serves as a major protein interaction network hub by potentially complexing with >100 distinct protein substrates, most of which function in highly conserved metabolic pathways. The interactome results were independently verified using a newly developed prokaryotic split luciferase complementation assay (SLCA) to detect MecA protein-protein interactionsin vivo. In addition, we further develop a new application of SLCA to supportin vivomeasurements of MecA relative protein binding affinities. SLCA results were independently verified using targeted coimmunoprecipitation assays, suggesting the general utility of this approach for prokaryotic protein-protein interaction studies. Our results indicate that MecA indeed regulates its interactome through both Clp-dependent proteolysis as well as through an as yet undefined proteolysis-independent mechanism that may affect more than half of its protein interactome. This suggests a significant aspect of MecA regulatory function still has yet to be discovered.

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“…Carlsson et al ( 1993 ) investigated the proteolytic ability 37 oral species and found that Peptostreptococcus micros ( Parvimonas micra ) and Fusobacterium nucleatum have a high capacity to degrade tripeptides and produce H 2 S. This behaviour is only found in opportunistic pathogens including Escherichia coli and Pseudomonas aeruginosa . Parvimonas micra is also the most common source of sepsis produced by GPAC (Higashi et al, 2022 ). The proteolytic capabilities of Parvimonas micra and Fusobacterium nucleatum are found in acute apical abscesses and seem to support mucosal dysbiosis also related to colon rectal cancer (George et al, 2016 ; Higashi et al, 2022 ; Siqueira & Rôças, 2009 ).…”

Section: Discussionmentioning

confidence: 99%

“…Parvimonas micra is also the most common source of sepsis produced by GPAC (Higashi et al, 2022 ). The proteolytic capabilities of Parvimonas micra and Fusobacterium nucleatum are found in acute apical abscesses and seem to support mucosal dysbiosis also related to colon rectal cancer (George et al, 2016 ; Higashi et al, 2022 ; Siqueira & Rôças, 2009 ). Interestingly, the combination of Parvimonas micra and Peptostreptococcus stomatis in combination with Fusobacterium nucleatum , and Solobacterium morei are potential biomarkers of colorectal cancer (Yu et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

Taxonomic abundance in primary and secondary root canal infections

et al. 2022

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Aim To evaluate the root canal microbiome composition in cases of primary and secondary apical periodontitis. Methodology Thirty‐nine samples from patients with primary root canal infections obtained before root canal treatment, and 40 samples obtained during root‐end resection procedures from previously filled cases with apical periodontitis were evaluated using 16S rRNA next‐generation sequencing analysis (NGS). Demographic and clinical factors included age, sex, infection type, percussion sensitivity, and presence of pain. Differences in abundances of genera were evaluated using Kruskal‐Wallis test. Alpha and beta diversity indices were calculated using mothur. The Shannon and Chao1 indices were used to measure alpha diversity. The Bray–Curtis dissimilarity was used to measure beta diversity. Differences in community composition were evaluated using analysis of similarity (ANOSIM) with Bonferroni correction for multiple comparisons. Results Significantly fewer operational taxonomic units values were observed from samples from secondary infections (p < .0001). While no significant differences were observed in the Chao 1 index between primary and secondary infections, the Shannon alpha diversity was significantly lower in secondary relative to primary infections (p = .008). Among samples, sex, age (adult vs. older adult), percussion sensitivity, and presence of pain all showed no significant effects on community composition via an analysis of similarity (ANOSIM). However, community composition was significantly different depending on whether the sample was from a primary or secondary infection (R = .051, p = .03). Nine microbial genera comprised the predominant taxa observed among samples (>3.3%) and included Parvimonas, Fusobacterium, Campylobacter, Arachnia, Eubacterium, Prevotella, Peptostreptococcus, Fretibacterirum, and Pseudoramibacter. Significantly greater relative abundances of Prevotella, Peptostreptococcus, Veillonella, Lactucaseibacillus, and Dialister were observed in primary infections. Conclusions Primary endodontic infections are more diverse than secondary infections. The microbial composition is not associated with the clinical manifestations of apical periodontitis.

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Development of the First Tractable Genetic System for Parvimonas micra, a Ubiquitous Pathobiont in Human Dysbiotic Disease

Abstract: Parvimonas micra is among the most highly enriched species at numerous sites of mucosal dysbiotic disease and is closely associated with numerous cancers. Despite this, little is known about P. micra pathobiology, which is directly attributable to its longstanding genetic intractability.

Cited by 10 publications

References 56 publications

Germinal center BCR maturation in appendicitis reveals a role for antigen-specific adaptive immune responses during disease

Germinal center BCR maturation in appendicitis reveals a role for antigen-specific adaptive immune responses during disease

Mass spectrometry and split luciferase complementation assays reveal the MecA protein interactome ofStreptococcus mutans

Taxonomic abundance in primary and secondary root canal infections

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