127: Characterization of the anti-hyperalgesic potential of probiotics from strains of Faecalibacterium

Topic: Basic Science
Subtopic: Pain category (ICD-11), Chronic primary visceral / pelvic pain, , , , , Animal imaging (includes Ca2+ imaging, fluorescence microscopy), , , ,

Background and Aims: Faecalibacterium has emerged as a promising next-generation probiotic, attracting considerable interest due to its anti-inflammatory properties. In addition, this strain has been shown to play an important role in the regulation of colonic hypersensitivity (CHS) in a chronic stress model induced by neonatal maternal separation (NMS). Moreover, patients suffering from irritable bowel syndrome (IBS) or inflammatory bowel disease (IBD) commonly exhibit intestinal dysbiosis, characterized by reduced microbial diversity and, in particular, a decreased abundance of Faecalibacterium genus. The present study investigates the therapeutic potential of this genus to modulate colonic sensitivity and the associated neuro-inhibitory mechanisms.

Methods: Several strains belonging to the genus Faecalibacterium were selected to investigate their anti-hyperalgesic effects in vivo using a non-inflammatory mouse model of CHS based on the NMS paradigm. C57Bl/6J mouse pups were subjected to NMS stress between postnatal days. Colonic sensitivity was assessed using a colorectal distension test coupled with a pressure sensor to monitor changes in intracolonic pressure. The neuro-inhibitory properties of different Faecalibacterium strains were evaluated in vitro using high-throughput and single-cell calcium imaging. Bacterial fractions were tested on an immortalized neuronal cell line as well as on primary cultures of neurons derived from dorsal root ganglia.

Results: This study demonstrated that several, but not all, tested strains of the Faecalibacterium genus significantly reduced NMS-induced CHS. Furthermore, analysis of neuronal responses to nociceptive stimulation with various algogenic substances revealed that culture supernatants from certain strains exerted a marked inhibitory effect on neuronal activation, highlighting their neuro-inhibitory potential.

Conclusions: The Faecalibacterium genus exhibits promising therapeutic potential for the treatment of CHS and associated pain in IBS and IBD during remission phases, owing to its anti-inflammatory, anti-hyperalgesic, and neuro-inhibitory properties. The diversity of strains and metabolites produced by this bacterium offers new perspectives for the development of innovative treatments for chronic abdominal pain.

References: 1. Sender, R., Fuchs, S., and Milo, R. (2016). Are We Really Vastly Outnumbered? Revisiting the Ratio of Bacterial to Host Cells in Humans. Cell 164, 337–340. https://doi.org/10.1016/j.cell.2016.01.013.
2. Sekirov, I., Russell, S.L., Antunes, L.C.M., and Finlay, B.B. (2010). Gut Microbiota in Health and Disease. Physiol. Rev. 90, 859–904. https://doi.org/10.1152/physrev.00045.2009.
3. Hugon, P., Dufour, J.-C., Colson, P., Fournier, P.-E., Sallah, K., and Raoult, D. (2015). A comprehensive repertoire of prokaryotic species identified in human beings. Lancet Infect. Dis. 15, 1211–1219. https://doi.org/10.1016/S1473-3099(15)00293-5.
4. Brüssow, H. (2020). Problems with the concept of gut microbiota dysbiosis. Microb. Biotechnol. 13, 423–434. https://doi.org/10.1111/1751-7915.13479.
5. Vangay, P., Ward, T., Gerber, J.S., and Knights, D. (2015). Antibiotics, Pediatric Dysbiosis, and Disease. Cell Host Microbe 17, 553–564. https://doi.org/10.1016/j.chom.2015.04.006.
6. Donohoe, D.R., Garge, N., Zhang, X., Sun, W., O’Connell, T.M., Bunger, M.K., and Bultman, S.J. (2011). The Microbiome and Butyrate Regulate Energy Metabolism and Autophagy in the Mammalian Colon. Cell Metab. 13, 517–526. https://doi.org/10.1016/j.cmet.2011.02.018.
7. Farhadi, A., Banan, A., Fields, J., and Keshavarzian, A. (2003). Intestinal barrier: An interface between health and disease. J. Gastroenterol. Hepatol. 18, 479–497. https://doi.org/10.1046/j.1440-1746.2003.03032.x.
8. Levy, O. (2007). Innate immunity of the newborn: basic mechanisms and clinical correlates. Nat. Rev. Immunol. 7, 379–390. https://doi.org/10.1038/nri2075.
9. Viola, M.F., Chavero-Pieres, M., Modave, E., Delfini, M., Stakenborg, N., Estévez, M.C., Fabre, N., Appeltans, I., Martens, T., Vandereyken, K., et al. (2023). Dedicated macrophages organize and maintain the enteric nervous system. Nature 618, 818–826. https://doi.org/10.1038/s41586-023-06200-7.
10. Cohen, S.P., Vase, L., and Hooten, W.M. (2021). Chronic pain: an update on burden, best practices, and new advances. The Lancet 397, 2082–2097. https://doi.org/10.1016/S0140-6736(21)00393-7.
11. LeBars, D. (2004). Physiologie de la douleur. EMC - Anesthésie-Réanimation 1: 2004th ed.
12. Defaye, M., Gervason, S., Altier, C., Berthon, J.-Y., Ardid, D., Filaire, E., and Carvalho, F.A. (2020). Microbiota: a novel regulator of pain. J. Neural Transm. 127, 445–465. https://doi.org/10.1007/s00702-019-02083-z.
13. Paré, P., Gray, J., Lam, S., Balshaw, R., Khorasheh, S., Barbeau, M., Kelly, S., and McBurney, C.R. (2006). Health-related quality of life, work productivity, and health care resource utilization of subjects with irritable bowel syndrome: Baseline results from logic (longitudinal outcomes study of gastrointestinal symptoms in Canada), a naturalistic study. Clin. Ther. 28, 1726–1735. https://doi.org/10.1016/j.clinthera.2006.10.010.
14. Marx, M., Maye, H., Abdelrahman, K., Hessler, R., Moschouri, E., Aslan, N., Godat, S., Nichita, C., Wiesel, P., Perez, L., et al. (2018). [Functional gastrointestinal disorders: update on the Rome IV criteria]. Rev. Med. Suisse 14, 1512–1516.
15. Martín, R., Laval, L., Chain, F., Miquel, S., Natividad, J., Cherbuy, C., Sokol, H., Verdu, E.F., Van Hylckama Vlieg, J., Bermudez-Humaran, L.G., et al. (2016). Bifidobacterium animalis ssp. lactis CNCM-I2494 Restores Gut Barrier Permeability in Chronically Low-Grade Inflamed Mice. Front. Microbiol. 7. https://doi.org/10.3389/fmicb.2016.00608.
16. Dionne, J., Ford, A.C., Yuan, Y., Chey, W.D., Lacy, B.E., Saito, Y.A., Quigley, E.M.M., and Moayyedi, P. (2018). A Systematic Review and Meta-Analysis Evaluating the Efficacy of a Gluten-Free Diet and a Low FODMAPS Diet in Treating Symptoms of Irritable Bowel Syndrome: Am. J. Gastroenterol. 113, 1290–1300. https://doi.org/10.1038/s41395-018-0195-4.
17. Uhlig, F., Grundy, L., Garcia-Caraballo, S., Brierley, S.M., Foster, S.J., and Grundy, D. (2020). Identification of a Quorum Sensing-Dependent Communication Pathway Mediating Bacteria-Gut-Brain Cross Talk. iScience 23, 101695. https://doi.org/10.1016/j.isci.2020.101695.
18. Marotto, D., Atzeni, F., Ardizzone, S., Monteleone, G., Giorgi, V., and Sarzi-Puttini, P. (2020). Extra-intestinal manifestations of inflammatory bowel diseases. Pharmacol. Res. 161, 105206. https://doi.org/10.1016/j.phrs.2020.105206.
19. Norton, C., Czuber‐Dochan, W., Artom, M., Sweeney, L., and Hart, A. (2017). Systematic review: interventions for abdominal pain management in inflammatory bowel disease. Aliment. Pharmacol. Ther. 46, 115–125. https://doi.org/10.1111/apt.14108.
20. Regueiro, M., Greer, J.B., and Szigethy, E. (2017). Etiology and Treatment of Pain and Psychosocial Issues in Patients With Inflammatory Bowel Diseases. Gastroenterology 152, 430-439.e4. https://doi.org/10.1053/j.gastro.2016.10.036.
21. Sokol, H., Seksik, P., Furet, J.P., Firmesse, O., Nion-Larmurier, I., Beaugerie, L., Cosnes, J., Corthier, G., Marteau, P., and Doré, J. (2009). Low counts of Faecalibacterium prausnitzii in colitis microbiota: Inflamm. Bowel Dis. 15, 1183–1189. https://doi.org/10.1002/ibd.20903.
22. Meynier, M., Daugey, V., Mallaret, G., Gervason, S., Meleine, M., Barbier, J., Aissouni, Y., Lolignier, S., Bonnet, M., Ardid, D., et al. (2024). Pasteurized Akkermansia muciniphila improves irritable bowel syndrome-like symptoms and related behavioral disorders in mice. Gut Microbes 16, 2298026. https://doi.org/10.1080/19490976.2023.2298026.
23. McCallum, S., Obata, Y., Fourli, E., Boeing, S., Peddie, C.J., Xu, Q., Horswell, S., Kelsh, R.N., Collinson, L., Wilkinson, D., et al. (2020). Enteric glia as a source of neural progenitors in adult zebrafish. eLife 9, e56086. https://doi.org/10.7554/eLife.56086.
24. Azpiroz, F., Bouin, M., Camilleri, M., Mayer, E.A., Poitras, P., Serra, J., and Spiller, R.C. (2007). Mechanisms of hypersensitivity in IBS and functional disorders. Neurogastroenterol. Motil. 19, 62–88. https://doi.org/10.1111/j.1365-2982.2006.00875.x.
25. Drewes, A.M., Olesen, A.E., Farmer, A.D., Szigethy, E., Rebours, V., and Olesen, S.S. (2020). Gastrointestinal pain. Nat. Rev. Dis. Primer 6, 1. https://doi.org/10.1038/s41572-019-0135-7.
26. Riba, A., Olier, M., Lacroix-Lamandé, S., Lencina, C., Bacquié, V., Harkat, C., Gillet, M., Baron, M., Sommer, C., Mallet, V., et al. (2017). Paneth Cell Defects Induce Microbiota Dysbiosis in Mice and Promote Visceral Hypersensitivity. Gastroenterology 153, 1594-1606.e2. https://doi.org/10.1053/j.gastro.2017.08.044.
27. Cenac, N., Altier, C., Chapman, K., Liedtke, W., Zamponi, G., and Vergnolle, N. (2008). Transient Receptor Potential Vanilloid-4 Has a Major Role in Visceral Hypersensitivity Symptoms. Gastroenterology 135, 937-946.e2. https://doi.org/10.1053/j.gastro.2008.05.024.
28. Cenac, N. (2013). Protease-Activated Receptors as Therapeutic Targets in Visceral Pain. Curr. Neuropharmacol. 11, 598–605. https://doi.org/10.2174/1570159X113119990039.
29. Cook, A.D., Christensen, A.D., Tewari, D., McMahon, S.B., and Hamilton, J.A. (2018). Immune Cytokines and Their Receptors in Inflammatory Pain. Trends Immunol. 39, 240–255. https://doi.org/10.1016/j.it.2017.12.003.
30. Pérez-Berezo, T., Pujo, J., Martin, P., Le Faouder, P., Galano, J.-M., Guy, A., Knauf, C., Tabet, J.C., Tronnet, S., Barreau, F., et al. (2017). Identification of an analgesic lipopeptide produced by the probiotic Escherichia coli strain Nissle 1917. Nat. Commun. 8, 1314. https://doi.org/10.1038/s41467-017-01403-9.
31. Martín, R., and Langella, P. (2019). Emerging Health Concepts in the Probiotics Field: Streamlining the Definitions. Front. Microbiol. 10, 1047. https://doi.org/10.3389/fmicb.2019.01047.
32. Plaza-Díaz, J., Ruiz-Ojeda, F., Vilchez-Padial, L., and Gil, A. (2017). Evidence of the Anti-Inflammatory Effects of Probiotics and Synbiotics in Intestinal Chronic Diseases. Nutrients 9, 555. https://doi.org/10.3390/nu9060555.
33. Ohland, C.L., and MacNaughton, W.K. (2010). Probiotic bacteria and intestinal epithelial barrier function. Am. J. Physiol.-Gastrointest. Liver Physiol. 298, G807–G819. https://doi.org/10.1152/ajpgi.00243.2009.
34. Miquel, S., Martín, R., Rossi, O., Bermúdez-Humarán, L., Chatel, J., Sokol, H., Thomas, M., Wells, J., and Langella, P. (2013). Faecalibacterium prausnitzii and human intestinal health. Curr. Opin. Microbiol. 16, 255–261. https://doi.org/10.1016/j.mib.2013.06.003.
35. Breyner, N.M., Michon, C., De Sousa, C.S., Vilas Boas, P.B., Chain, F., Azevedo, V.A., Langella, P., and Chatel, J.M. (2017). Microbial Anti-Inflammatory Molecule (MAM) from Faecalibacterium prausnitzii Shows a Protective Effect on DNBS and DSS-Induced Colitis Model in Mice through Inhibition of NF-κB Pathway. Front. Microbiol. 8. https://doi.org/10.3389/fmicb.2017.00114.
36. Sokol, H., Pigneur, B., Watterlot, L., Lakhdari, O., Bermúdez-Humarán, L.G., Gratadoux, J.-J., Blugeon, S., Bridonneau, C., Furet, J.-P., Corthier, G., et al. (2008). Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients. Proc. Natl. Acad. Sci. 105, 16731–16736. https://doi.org/10.1073/pnas.0804812105.
37. Meleine, M., Boudieu, L., Gelot, A., Muller, E., Lashermes, A., Matricon, J., Silberberg, C., Theodorou, V., Eschalier, A., Ardid, D., et al. (2016). Comparative effects of α2δ-1 ligands in mouse models of colonic hypersensitivity. World J. Gastroenterol. 22, 7111. https://doi.org/10.3748/wjg.v22.i31.7111.
38. Sakamoto, M., Sakurai, N., Tanno, H., Iino, T., Ohkuma, M., and Endo, A. (2022). Genome-based, phenotypic and chemotaxonomic classification of Faecalibacterium strains: proposal of three novel species Faecalibacterium duncaniae sp. nov., Faecalibacterium hattorii sp. nov. and Faecalibacterium gallinarum sp. nov. Int. J. Syst. Evol. Microbiol. 72. https://doi.org/10.1099/ijsem.0.005379.
39. Quévrain, E., Maubert, M.-A., Sokol, H., Devreese, B., and Seksik, P. (2016). The presence of the anti-inflammatory protein MAM, from Faecalibacterium prausnitzii , in the intestinal ecosystem. Gut 65, 882.1-882. https://doi.org/10.1136/gutjnl-2015-311094.
40. Miquel, S., Martín, R., Lashermes, A., Gillet, M., Meleine, M., Gelot, A., Eschalier, A., Ardid, D., Bermúdez-Humarán, L.G., Sokol, H., et al. (2016). Anti-nociceptive effect of Faecalibacterium prausnitzii in non-inflammatory IBS-like models. Sci. Rep. 6, 19399. https://doi.org/10.1038/srep19399.
41. Auger, S., Kropp, C., Borras-Nogues, E., Chanput, W., Andre-Leroux, G., Gitton-Quent, O., Benevides, L., Breyner, N., Azevedo, V., Langella, P., et al. (2022). Intraspecific Diversity of Microbial Anti-Inflammatory Molecule (MAM) from Faecalibacterium prausnitzii. Int. J. Mol. Sci. 23, 1705. https://doi.org/10.3390/ijms23031705.
42. Le Berre‐Scoul, C., Chevalier, J., Oleynikova, E., Cossais, F., Talon, S., Neunlist, M., and Boudin, H. (2017). A novel enteric neuron–glia coculture system reveals the role of glia in neuronal development. J. Physiol. 595, 583–598. https://doi.org/10.1113/JP271989.
43. Larauche, M., Mulak, A., Yuan, P.-Q., Kanauchi, O., and Taché, Y. (2012). Stress-induced visceral analgesia assessed non-invasively in rats is enhanced by prebiotic diet. World J. Gastroenterol. 18, 225–236. https://doi.org/10.3748/wjg.v18.i3.225.
44. Gervason, S., Meleine, M., Lolignier, S., Meynier, M., Daugey, V., Birer, A., Aissouni, Y., Berthon, J.-Y., Ardid, D., Filaire, E., et al. (2023). Antihyperalgesic properties of gut microbiota: Parabacteroides distasonis as a new probiotic strategy to alleviate chronic abdominal pain. Pain. https://doi.org/10.1097/j.pain.0000000000003075.
45. Caillaud, M., Le Dréan, M.E., De-Guilhem-de-Lataillade, A., Le Berre-Scoul, C., Montnach, J., Nedellec, S., Loussouarn, G., Paillé, V., Neunlist, M., and Boudin, H. (2023). A functional network of highly pure enteric neurons in a dish. Front. Neurosci. 16, 1062253. https://doi.org/10.3389/fnins.2022.1062253.
46. Quévrain, E., Maubert, M.A., Michon, C., Chain, F., Marquant, R., Tailhades, J., Miquel, S., Carlier, L., Bermúdez-Humarán, L.G., Pigneur, B., et al. (2016). Identification of an anti-inflammatory protein from Faecalibacterium prausnitzii , a commensal bacterium deficient in Crohn’s disease. Gut 65, 415–425. https://doi.org/10.1136/gutjnl-2014-307649.
47. Gervason, S., Meleine, M., Lolignier, S., Meynier, M., Daugey, V., Birer, A., Aissouni, Y., Berthon, J.-Y., Ardid, D., Filaire, E., et al. (2024). Antihyperalgesic properties of gut microbiota: Parabacteroides distasonis as a new probiotic strategy to alleviate chronic abdominal pain. Pain 165, e39–e54. https://doi.org/10.1097/j.pain.0000000000003075.
48. Petitfils, C., Maurel, S., Payros, G., Hueber, A., Agaiz, B., Gazzo, G., Marrocco, R., Auvray, F., Langevin, G., Motta, J.-P., et al. (2023). Identification of bacterial lipopeptides as key players in IBS. Gut 72, 939–950. https://doi.org/10.1136/gutjnl-2022-328084.
49. Defaye, M., Gervason, S., Altier, C., Berthon, J.-Y., Ardid, D., Filaire, E., and Carvalho, F.A. (2020). Microbiota: a novel regulator of pain. J. Neural Transm. 127, 445–465. https://doi.org/10.1007/s00702-019-02083-z.
50. Defaye, M., Abdullah, N.S., Iftinca, M., Hassan, A., Agosti, F., Zhang, Z., Cumenal, M., Zamponi, G.W., and Altier, C. (2022). Gut-innervating TRPV1+ Neurons Drive Chronic Visceral Pain via Microglial P2Y12 Receptor. Cell. Mol. Gastroenterol. Hepatol. 13, 977–999. https://doi.org/10.1016/j.jcmgh.2021.12.012.

Presenting Author: PERAUT CAMILLE
Submitting Author: Camille PERAUT

Senior Author: PERAUT CAMILLE

IASP 2026 Global Year theme of Neuropathic Pain: Yes

• CARVALHO FREDERIC: Research Director INSERM - Neuro-Dol
• CHATEL Jean-Marc: Director of Research - MICALIS