P300 COUNTERBALANCING DYSBIOSIS IN CROHN'S DISEASE: FAECALIBACTERIUM PRAUSNITZII, A MAJOR COMMENSAL BACTERIUM, EXHIBITS IN VITRO AND IN VIVO ANTI-INFLAMMATORY EFFECTS

Abstract BackgroundThe commensal bacterium Faecalibacterium prausnitzii plays a key role in inflammatory bowel disease (IBD) pathogenesis and serves as a general health biomarker in humans. However, the host molecular mechanisms that underlie its anti-inflammatory effects remain unknown.MethodsA transcriptomic approach on human intestinal epithelial cells (HT-29) that were stimulated with TNF-α and exposed to F. prausnitzii culture supernatant (SN) was used. Modulation of the most upregulated gene after F. prausnitzii SN contact was validated both in vitro and in vivo.ResultsF. prausnitzii SN upregulates the expression of Dact3, a gene linked to the Wnt/JNK pathway. Interestingly, when we silenced Dact3 expression, the effect of F. prausnitzii SN was lost. Butyrate was identified as the F. prausnitzii effector responsible for Dact3 modulation. Dact3 upregulation was also validated in vivo in both healthy and inflamed mice treated with either F. prausnitzii SN or the live bacteria, respectively. Finally, we demonstrated by colon transcriptomics that gut microbiota directly influences Dact3 expression.ConclusionsOur results provide new clues about the host molecular mechanisms involved in the anti-inflammatory effects of the beneficial commensal bacterium F. prausnitzii.*Contributed equally to this work

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P020 MAM, an anti-inflammatory protein derived from Faecalibacterium prausnitzii as a biomarker in Crohn’s disease?

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjz203.149 ◽

2020 ◽

Vol 14 (Supplement_1) ◽

pp. S140-S141

Author(s):

P McLellan ◽

A Lavelle ◽

L Brot ◽

M Straube ◽

L de Sordi ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Predictive Value ◽

Healthy Controls ◽

Relative Expression ◽

Faecalibacterium Prausnitzii ◽

Inflammatory Protein ◽

Faecal Samples ◽

Anti Inflammatory

Abstract Background The intestinal microbiota of patients with Crohn’s disease (CD) is characterised by a specific dysbiosis, including the loss of Faecalibacterium prausnitzii (F. prau), a major bacterium with anti-inflammatory properties. MAM protein (microbiotia anti-inflammatory molecule) produced by F. prau has recently been identified as partially carrying the anti-inflammatory activity of this bacterium. The description of microbiota for diagnostic purposes in CD is currently difficult to achieve. MAM could be a biomarker of dysbiosis. Our goal was to study the expression of MAM in the intestinal ecosystem and to evaluate its interest as a biomarker. Methods An in silico approach was used to determine primers of MAM gene sequences specific to two currently known phylogroups (I and II) of F. prau. Faecal samples were collected from a monocentric cohort of CD patients in flare (n = 24) and in remission (n = 24) and from healthy controls (n = 12). To measure MAM expression in vivo in the microbiota, we extracted total RNA and DNA from each sample and performed quantitative PCR (qPCR) with MAM primers (I and II), F. prau primers and universal bacterial primers. We compared the relative expression (ΔCt) of MAM mRNA and DNA with qPCR to the overall amount of bacteria in faecal samples. Results MAM analyses of F. prau phylogroup II were the most informative. We observed that the relative expression of MAM mRNA was statistically lower in active patients 5.84 × 10−5 [95% CI 3.8 × 10−6–2.1 × 10−3] and in remission 5.36 × 10−5 [95% CI 2.5 × 10−8–1.9 × 10−2] compared with healthy controls 5.42 × 10−4 [95% CI 8.5 × 10−5–9.1 × 10−2], (p = 0.004). There was no difference in ΔCt between flare and remission in CD patients. The most relevant relative expression threshold was 1.6 × 10−4. At this threshold, sensitivity was 81%, specificity 76%, positive predictive value 94% and negative predictive value 47%. In multivariate analysis, the relative expression of MAM was an independently associated with the diagnosis of CD; OR 2.961 [95% CI 2.026–4.831] (p = 0.003). Using relative expression of MAM DNA, we found a significant decrease in the flare group compared with remission patients and controls. We found a negative correlation of MAM DNA expression with CRP (p = 0.0053) and HBI (p = 0.025). Conclusion Our results showed that the loss of MAM RNA expression is observed in CD patients. Moreover, MAM DNA detection was associated with CD activity parameters. This opens up the prospect of an ecological biomarker of CD based on the expression of molecules from the intestinal ecosystem. These results will need to be validated in larger independent cohorts and their specificities compared with other pathological situations.

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Creatine Supplementation for Patients with Inflammatory Bowl Diseases: A Scientific Rationale for a Clinical Trial

Nutrients ◽

10.3390/nu13051429 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1429

Author(s):

Theo Wallimann ◽

Caroline H. T. Hall ◽

Sean P. Colgan ◽

Louise E. Glover

Keyword(s):

Ulcerative Colitis ◽

Clinical Trial ◽

Crohn’S Disease ◽

Pilot Study ◽

Crohn's Disease ◽

Single Case ◽

Initial Period ◽

Creatine Supplementation

Based on theoretical considerations, experimental data with cells in vitro, animal studies in vivo, as well as a single case pilot study with one colitis patient, a consolidated hypothesis can be put forward, stating that “oral supplementation with creatine monohydrate (Cr), a pleiotropic cellular energy precursor, is likely to be effective in inducing a favorable response and/or remission in patients with inflammatory bowel diseases (IBD), like ulcerative colitis and/or Crohn’s disease”. A current pilot clinical trial that incorporates the use of oral Cr at a dose of 2 × 7 g per day, over an initial period of 2 months in conjunction with ongoing therapies (NCT02463305) will be informative for the proposed larger, more long-term Cr supplementation study of 2 × 3–5 g of Cr per day for a time of 3–6 months. This strategy should be insightful to the potential for Cr in reducing or alleviating the symptoms of IBD. Supplementation with chemically pure Cr, a natural nutritional supplement, is well tolerated not only by healthy subjects, but also by patients with diverse neuromuscular diseases. If the outcome of such a clinical pilot study with Cr as monotherapy or in conjunction with metformin were positive, oral Cr supplementation could then be used in the future as potentially useful adjuvant therapeutic intervention for patients with IBD, preferably together with standard medication used for treating patients with chronic ulcerative colitis and/or Crohn’s disease.

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Sonographic features of colonic Crohn's disease: Comparison of in vivo and in vitro studies

Journal of Clinical Ultrasound ◽

10.1002/jcu.1870180304 ◽

1990 ◽

Vol 18 (3) ◽

pp. 161-166 ◽

Cited By ~ 20

Author(s):

Bernd Limberg

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

In Vitro Studies

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The commensal bacterium Rothia mucilaginosa has anti-inflammatory properties in vitro and in vivo, and negatively correlates with sputum pro-inflammatory markers in chronic airway disease

10.1183/13993003.congress-2021.pa873 ◽

2021 ◽

Author(s):

Charlotte Rigauts ◽

Juliana Aizawa ◽

Steven Taylor ◽

Geraint Rogers ◽

Matthias Govaerts ◽

...

Keyword(s):

Inflammatory Markers ◽

Airway Disease ◽

Commensal Bacterium ◽

Chronic Airway Disease ◽

Rothia Mucilaginosa ◽

Anti Inflammatory ◽

Chronic Airway

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Mesenchymal Stromal Cell Therapy in Crohn's Disease

Digestive Diseases ◽

10.1159/000449091 ◽

2017 ◽

Vol 35 (1-2) ◽

pp. 115-122 ◽

Cited By ~ 15

Author(s):

Geoffrey M. Forbes

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Adult Stem Cells ◽

Hla Class I ◽

Haematopoietic Stem Cell ◽

Dose Frequency ◽

Serious Adverse Events ◽

Considerable Work

Background: Mesenchymal stromal cells (MSC) are multipotent adult stem cells with immunomodulatory properties. They uniquely express HLA class I antigen at a low level, and do not express HLA class II. Hence, for allogeneic administration, donor to recipient matching is not required; yet a prolonged chimeric state does not occur. Contrary to haematopoietic stem cell transplantation, cytotoxic drug therapy is not required to harvest, or administer, cells. Key Messages: MSC are obtained from marrow, adipose tissue or placenta. In our centre, MSC are isolated from a 10 ml donor marrow aspirate, by virtue of their adherence to plastic. They are expanded in culture, cryopreserved, and subjected to strict quality controls before release for intravenous administration. These activities occur in a dedicated, nationally accredited, laboratory. Initial observations of allogeneic MSC efficacy were in graft-versus-host disease. Both autologous and allogeneic MSC have since been evaluated in biologic refractory luminal and fistulising Crohn's disease (CD). Data from early-phase studies have suggested efficacy for luminal disease when allogeneic MSC were given intravenously and also suggested efficacy for fistulising disease when either allogeneic or autologous MSC were administered into fistulas. MSC treatment is not reported to have caused serious adverse events. Although in vitro criteria for defining MSC exist, a major challenge lies in how to define MSC for clinical use. MSC function in vivo is likely to be dependent upon donor immunological characteristics, and widely varying manufacturing processes between laboratories. MSC dose, frequency of administration, stage of disease, and presence of concomitant immunosuppression also require to be defined. Conclusions: MSC therapy may have future utility in CD, but considerable work is first required to determine appropriate phenotypic and functional characteristics of administered cells.

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Identification of an anti-inflammatory protein fromFaecalibacterium prausnitzii, a commensal bacterium deficient in Crohn’s disease

Gut ◽

10.1136/gutjnl-2014-307649 ◽

2015 ◽

Vol 65 (3) ◽

pp. 415-425 ◽

Cited By ~ 267

Author(s):

E Quévrain ◽

M A Maubert ◽

C Michon ◽

F Chain ◽

R Marquant ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Commensal Bacterium ◽

Inflammatory Protein ◽

Anti Inflammatory

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The effect of marine oil-derived n-3 fatty acids on transepithelial calcium transport in Caco-2 cell models of healthy and inflamed intestines

British Journal Of Nutrition ◽

10.1017/s0007114507201758 ◽

2007 ◽

Vol 97 (2) ◽

pp. 281-288 ◽

Cited By ~ 4

Author(s):

Jennifer Gilman ◽

Kevin D. Cashman

Keyword(s):

Fatty Acids ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Human Subjects ◽

Healthy Human ◽

Marine Oil ◽

Ca Transport ◽

Tnf Α

Marine oil-derived n-3 fatty acids have been shown to stimulate intestinal Ca absorption in animal studies, but the effects of such fatty acids on Ca absorption in human subjects are relatively unknown. In particular, n-3 fatty acids may be of therapeutic value for some Crohn's disease patients who experience Ca malabsorption. Therefore, the aim of the present study was to investigate the effect of 20 : 5n-3 and 22 : 6n-3 on transepithelial Ca transport across monolayers of healthy Caco-2 cells as well as of TNF-α-treated Caco-2 cells (an in vitro model of Crohn's disease). Caco-2 cells were seeded onto permeable filter supports and allowed to differentiate into monolayers, which were treated with 80 μm-20 : 5n-3, 80 μm-22 : 6n-3, or 40 μm-20 : 5n-3+40 μm-22 : 6n-3 for 6 or 8 d, with or without co-treatment with TNF-α (10 ng/ml) (n 11–15 monolayers per treatment). On day 16, transepithelial and transcellular transport of 45Ca and fluorescein transport (a marker of paracellular diffusion) were measured. Treatment of healthy and inflamed Caco-2 cells with 20 : 5n-3, 22 : 6n-3 and both fatty acids combined for 8 d significantly (P < 0·005–0·01) increased total transepithelial Ca transport compared with that in control, effects which were mediated by an enhanced rate of transcellular Ca transport. The effects of n-3 fatty acids on Ca absorption after 6 d were less clear-cut. In conclusion, the present in vitro findings highlight the need to investigate the effect of marine oil-based n-3 fatty acids on Ca absorption in vivo in studies of healthy human subjects as well as of Crohn's disease patients.

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Co-Culture with Bifidobacterium catenulatum Improves the Growth, Gut Colonization, and Butyrate Production of Faecalibacterium prausnitzii: In Vitro and In Vivo Studies

Microorganisms ◽

10.3390/microorganisms8050788 ◽

2020 ◽

Vol 8 (5) ◽

pp. 788 ◽

Cited By ~ 2

Author(s):

Heejung Kim ◽

Yunju Jeong ◽

Sini Kang ◽

Hyun Ju You ◽

Geun Eog Ji

Keyword(s):

Culture Supernatant ◽

Short Chain ◽

In Vivo Studies ◽

Fatty Acid Production ◽

Gut Colonization ◽

Faecalibacterium Prausnitzii ◽

Anti Inflammatory ◽

Butyrate Production

Faecalibacterium prausnitzii is a major commensal bacterium in the human gut. It produces short-chain fatty acids that promote intestinal health. However, the bacterium is extremely oxygen-sensitive, making it difficult to develop as a probiotic. To facilitate practical application of F. prausnitzii, we investigated factors that affect its growth and mammalian gut colonization. We evaluated cross-feeding interactions between F. prausnitzii and seven Bifidobacterium strains, and the anti-inflammatory properties of bacterial metabolites produced in co-culture, in vitro and in vivo. Co-culture of F. prausnitzii and Bifidobacterium catenulatum, with fructooligosaccharides as an energy source, resulted in the greatest viable cell-count and butyrate production increases. Further, the co-culture supernatant reduced the amount of proinflammatory cytokines produced by HT-29 cells and RAW 264.7 macrophages, an effect that was similar to that of butyrate. Furthermore, feeding mice both Faecalibacterium and Bifidobacterium enhanced F. prausnitzii gut colonization. Finally, feeding the co-culture supernatant decreased interleukin 8 levels in the colon and increased butyrate levels in the cecum in the dextran sodium sulfate-induced colitis mouse model. These observations indicate that the Faecalibacterium-Bifidobacterium co-culture exerts an anti-inflammatory effect by promoting F. prausnitzii survival and short-chain fatty acid production, with possible implications for the treatment of inflammatory bowel disease.

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