Commensal gut bacteria modulate phosphorylation-dependent PPARγ transcriptional activity in human intestinal epithelial cells

In healthy subjects, the intestinal microbiota interacts with the host’s epithelium, regulating gene expression to the benefit of both, host and microbiota. The underlying mechanisms remain poorly understood, however. Although many gut bacteria are not yet cultured, constantly growing culture collections have been established. We selected 57 representative commensal bacterial strains to study bacteria-host interactions, focusing on PPARγ, a key nuclear receptor in colonocytes linking metabolism and inflammation to the microbiota. Conditioned media (CM) were harvested from anaerobic cultures and assessed for their ability to modulate PPARγ using a reporter cell line. Activation of PPARγ transcriptional activity was linked to the presence of butyrate and propionate, two of the main metabolites of intestinal bacteria. Interestingly, some stimulatory CMs were devoid of these metabolites. A Prevotella and an Atopobium strain were chosen for further study, and shown to up-regulate two PPARγ-target genes, ANGPTL4 and ADRP. The molecular mechanisms of these activations involved the phosphorylation of PPARγ through ERK1/2. The responsible metabolites were shown to be heat sensitive but markedly diverged in size, emphasizing the diversity of bioactive compounds found in the intestine. Here we describe different mechanisms by which single intestinal bacteria can directly impact their host’s health through transcriptional regulation.

[1]  G. Zhou,et al.  Insulin- and Mitogen-activated Protein Kinase-mediated Phosphorylation and Activation of Peroxisome Proliferator-activated Receptor γ* , 1996, The Journal of Biological Chemistry.

[2]  R. Hughes,et al.  Fecal Water as a Non-Invasive Biomarker in Nutritional Intervention: Comparison of Preparation Methods and Refinement of Different Endpoints , 2007, Nutrition and cancer.

[3]  J. Gustafsson,et al.  Decreased Fat Storage by Lactobacillus Paracasei Is Associated with Increased Levels of Angiopoietin-Like 4 Protein (ANGPTL4) , 2010, PloS one.

[4]  U. Vogel,et al.  Polymorphisms in NF-κB, PXR, LXR, PPARγ and risk of inflammatory bowel disease. , 2011, World journal of gastroenterology.

[5]  M. Mohammadi,et al.  Faculty Opinions recommendation of Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. , 2018, Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature.

[6]  J. Doré,et al.  Identification of NF-κB Modulation Capabilities within Human Intestinal Commensal Bacteria , 2011, Journal of biomedicine & biotechnology.

[7]  G. Macfarlane,et al.  Short chain fatty acids in human large intestine, portal, hepatic and venous blood. , 1987, Gut.

[8]  F. Bäckhed,et al.  Gut microbiota regulates bile acid metabolism by reducing the levels of tauro-beta-muricholic acid, a naturally occurring FXR antagonist. , 2013, Cell metabolism.

[9]  Malcolm J. McConville,et al.  MR1 presents microbial vitamin B metabolites to MAIT cells , 2012, Nature.

[10]  Wei Sun,et al.  The microbiome and butyrate regulate energy metabolism and autophagy in the mammalian colon. , 2011, Cell metabolism.

[11]  Patrice D Cani,et al.  Role of gut microflora in the development of obesity and insulin resistance following high-fat diet feeding. , 2008, Pathologie-biologie.

[12]  N. Cerf-Bensussan,et al.  The immune system and the gut microbiota: friends or foes? , 2010, Nature Reviews Immunology.

[13]  Philippe Chavatte,et al.  PPARγ as a new therapeutic target in inflammatory bowel diseases , 2006, Gut.

[14]  M. Lazar,et al.  A novel therapy for colitis utilizing PPAR-gamma ligands to inhibit the epithelial inflammatory response. , 1999, The Journal of clinical investigation.

[15]  M. Lazar,et al.  Modulating nuclear receptor function: may the phos be with you. , 1999, The Journal of clinical investigation.

[16]  P. Bork,et al.  Richness of human gut microbiome correlates with metabolic markers , 2013, Nature.

[17]  S. Ganguly,et al.  Gut microbiota in health and disease , 2019, APIK Journal of Internal Medicine.

[18]  J. Doré,et al.  ANGPTL4 expression induced by butyrate and rosiglitazone in human intestinal epithelial cells utilizes independent pathways. , 2013, American journal of physiology. Gastrointestinal and liver physiology.

[19]  Anders K. Haakonsson,et al.  Short-Chain Fatty Acids Stimulate Angiopoietin-Like 4 Synthesis in Human Colon Adenocarcinoma Cells by Activating Peroxisome Proliferator-Activated Receptor γ , 2013, Molecular and Cellular Biology.

[20]  Samuel Singer,et al.  Differentiation and reversal of malignant changes in colon cancer through PPARγ , 1998, Nature Medicine.

[21]  C. Huttenhower,et al.  Expansion of intestinal Prevotella copri correlates with enhanced susceptibility to arthritis , 2013, eLife.

[22]  M. Moeschberger,et al.  New Bacterial Species Associated with Chronic Periodontitis , 2003, Journal of dental research.

[23]  生島 仁史,et al.  放射線治療--state of the art and in future , 2009 .

[24]  P. Sansonetti,et al.  A Crypt-Specific Core Microbiota Resides in the Mouse Colon , 2012, mBio.

[25]  R. Wolff,et al.  PPARγ and Colon and Rectal Cancer: Associations with Specific Tumor Mutations, Aspirin, Ibuprofen and Insulin-Related Genes (United States) , 2006, Cancer Causes & Control.

[26]  Christine Dreyer,et al.  Control of the peroxisomal β-oxidation pathway by a novel family of nuclear hormone receptors , 1992, Cell.

[27]  V. Annese,et al.  PPARγ in Inflammatory Bowel Disease , 2012, PPAR research.

[28]  K. Morimura,et al.  Peroxisome proliferator activated receptor gamma in colonic epithelial cells protects against experimental inflammatory bowel disease. , 2006, Gut.

[29]  Jonathan R. Brestoff,et al.  Commensal bacteria at the interface of host metabolism and the immune system , 2013, Nature Immunology.

[30]  J. V. Vanden Heuvel,et al.  Modulation of PPAR activity via phosphorylation. , 2007, Biochimica et biophysica acta.

[31]  M. Icaza-Chávez,et al.  Gut microbiota in health and disease , 2013 .

[32]  S. Ferrari,et al.  Author contributions , 2021 .

[33]  Jiahuai Han,et al.  Pro-inflammatory Cytokines and Environmental Stress Cause p38 Mitogen-activated Protein Kinase Activation by Dual Phosphorylation on Tyrosine and Threonine (*) , 1995, The Journal of Biological Chemistry.

[34]  J. Cummings Microbial Digestion of Complex Carbohydrates in Man , 1984, Proceedings of the Nutrition Society.

[35]  J. Gustafsson,et al.  Enterococcus faecalis from newborn babies regulate endogenous PPARγ activity and IL-10 levels in colonic epithelial cells , 2008, Proceedings of the National Academy of Sciences.

[36]  X. Thuru,et al.  PPARgamma as a new therapeutic target in inflammatory bowel diseases. , 2006, Gut.

[37]  J. Doré,et al.  Commensal Streptococcus salivarius Modulates PPARγ Transcriptional Activity in Human Intestinal Epithelial Cells , 2015, PloS one.

[38]  Ting Wang,et al.  The gut microbiota as an environmental factor that regulates fat storage. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[39]  W. Wahli,et al.  Fat poetry: a kingdom for PPAR gamma. , 2007, Cell research.

[40]  S. Lalevée,et al.  Phosphorylation control of nuclear receptors. , 2010, Methods in molecular biology.

[41]  J. Auwerx,et al.  The Organization, Promoter Analysis, and Expression of the Human PPARγ Gene* , 1997, The Journal of Biological Chemistry.

[42]  Jeffrey I. Gordon,et al.  Mechanisms underlying the resistance to diet-induced obesity in germ-free mice , 2007, Proceedings of the National Academy of Sciences.

[43]  M. Lazar,et al.  Transcriptional Activation by Peroxisome Proliferator-activated Receptor γ Is Inhibited by Phosphorylation at a Consensus Mitogen-activated Protein Kinase Site* , 1997, The Journal of Biological Chemistry.

[44]  S. Pettersson,et al.  Commensal anaerobic gut bacteria attenuate inflammation by regulating nuclear-cytoplasmic shuttling of PPAR-γ and RelA , 2004, Nature Immunology.

[45]  J. Gordon,et al.  How host-microbial interactions shape the nutrient environment of the mammalian intestine. , 2002, Annual review of nutrition.

[46]  M. Delday,et al.  Microbes and microbial effector molecules in treatment of inflammatory disorders , 2012, Immunological reviews.

[47]  J. Doré,et al.  Human intestinal metagenomics: state of the art and future. , 2013, Current opinion in microbiology.

[48]  C. Marshall,et al.  Specificity of receptor tyrosine kinase signaling: Transient versus sustained extracellular signal-regulated kinase activation , 1995, Cell.

[49]  Jeremy K. Nicholson,et al.  Gut microbiota: a potential new territory for drug targeting , 2008, Nature Reviews Drug Discovery.

[50]  W. Wahli,et al.  Fat poetry: a kingdom for PPARγ , 2007, Cell Research.

[51]  B. Spiegelman,et al.  Loss-of-function mutations in PPAR gamma associated with human colon cancer. , 1999, Molecular cell.

[52]  B. Spiegelman,et al.  Loss-of-Function Mutations in PPARγ Associated with Human Colon Cancer , 1999 .

[53]  K. Morimura,et al.  Peroxisome proliferator activated receptor γ in colonic epithelial cells protects against experimental inflammatory bowel disease , 2005, Gut.

[54]  A. Nakajima,et al.  PPARγ and inflammatory bowel disease: a new therapeutic target for ulcerative colitis and Crohn's disease , 2001 .

[55]  V. Mathan,et al.  Vitamin B12 synthesis by human small intestinal bacteria , 1980, Nature.

[56]  Joël Doré,et al.  Butyrate Produced by Commensal Bacteria Potentiates Phorbol Esters Induced AP-1 Response in Human Intestinal Epithelial Cells , 2012, PloS one.