and undernutrition Laure Bindels, PhD Copenhagen September 7, 2016 - PowerPoint PPT Presentation

NuGO week 2016 Role of the gut microbiota in over- and undernutrition Laure Bindels, PhD Copenhagen September 7, 2016

Adapted from Delzenne et al ., Nat Rev Endocrinol 2011

Outline 1. Gut microbiota as a nutritional target 2. Metabolic disorders associated with obesity 3. Metabolic disorders associated with cancer 4. Gut microbiota in alcohol-dependent patients

The gut microbiota 40 000 000 000 000 microbes 30 000 000 000 000 human cells for a 'reference man' (70 kilograms, 20 – 30 years old and 1.7 meters tall) Numbers from Sender et al, preprint on bioRxiv, 2016. Neish , Gastroenterology 2009

Gut microbiota-host crosstalk Delzenne et al ., Nat Rev Endocrinol 2011

Prebiotics FMT Probiotics Bacteriocins i.e. inulin-type fructans i.e. lactobacilli Antibiotics Gut microbiota Experimental tools to study our microbial partners Adapted from Bindels & Delzenne , Int J Biochem Cell Biol 2013

Prebiotics Beneficial physiological effects Gibson & Roberfroid, J Nutr 1995

Future research on prebiotics Figure 1 Current and proposed definitions for the concept of prebiotics Bindels et al, Nat Rev Gastroenterol Hepatol 2015

Resistant starches Resistant starches (RS) include all starch and starch degradation products not absorbed in the small intestine of healthy individuals. Asp, Trends Food Sci Technol, 1992; Birt et al., Adv Nutr 2013; Martinez et al, Plos ONE 2010.

Outline 1. Gut microbiota as a nutritional target 2. Metabolic disorders associated with obesity 3. Metabolic disorders associated with cancer 4. Gut microbiota in alcohol-dependent patients

Cancer cachexia Bindels & Thissen, Clin Nutr Exp 2015

Cancer cachexia • Up to 80% of cancer patients, depending of the tumor site • Reduces quality and length of life • May be a cause of cancer therapy discontinuation • No valid treatment Giacometti, Walking man Fearon et al. , Cell Metab 2012; Fearon et al., Nat Rev Oncol 2013; Argiles et al, Nat Rev Cancer 2014.

A microbial signature in cancer cachexia Community-wide approach to characterize the gut microbiota in two mouse models of cancer cachexia C26 BaF BaF3 cells with Bcr-Abl Bindels et al, The ISME J 2016

A microbial signature in cancer cachexia BaF C26 16S rRNA genes from the caecal microbiota analysed by Illumina MiSeq. Logarythmic LDA score. ↑ Enterobacteriaceae ↑ Parabacteroides goldsteinii ↓ Lactobacilli With Inès Martinez and Jens Walter Bindels et al, The ISME J 2016

… independent of the food intake * 125 11 ** (% initial food intake) Enterobacteriaceae *** *** 100 10 Daily food intake (log 10 [cells/g]) 75 9 # CT (BaF) 50 8 # BaF * CT (DR) 25 7 DR 0 6 CT(BaF) BaF CT(DR) DR 0 2 4 6 8 10 12 14 Days after BAF injection * *** *** *** 11 Parabacteroides goldsteinii ASF519 (log 10 [AU/g]) 10 Lactobacillus spp. (log 10 [cells/g]) 10 9 9 8 8 7 CT(BaF) BaF CT(DR) DR CT(BaF) BaF CT(DR) DR Bindels et al, The ISME J 2016

BaF intact BaF3/w ell B c r - A b l control 1.0  10 7 propionate 2mM Number of 1 . 0 5.0  10 6 § * * Acetate ls e n ) v io 0 . 5 0 le s A s N re 0 24 48 72 R p m x e e tiv la (re N D Propionate 0 . 0 CT BaF3 aF3-IT Butyrate ITF Acetate P r o p i o n a t e Propionate Butyrate § 6 0 Acetate 4 0 M µ Propionate 2 0 Butyrate 0 BaF CT F3- Bindels et al, Br J Cancer 2012; Bindels*, Neyrinck* et al, Plos ONE 2015.

Selected synbiotic approach D0 D1 D13 † L. reuteri 100-23 ITF Bcr-Abl-expressing BaF3 cells With Bruno Pot & Corinne Grangette Bindels et al, The ISME J 2016

BaF 16S rRNA genes from the caecal microbiota analysed by Illumina MiSeq. LEfSe cladogram. Bindels et al, The ISME J 2016

Benefits of the synbiotic approach Bcr-Abl BaF 0.50 * # 0.45 * (relative expression) 1.0 (% body weight) 0.40 Organ weight mRNA levels # 0.35 0.20 * $ 0.5 * 0.15 0.10 0.05 ND 0.0 0.00 CT BaF BaF-LrI CT BaF BaF-LrI CT BaF BaF-LrI tibialis gastrocnemius Morbidity score Survival 5 1.2 BaF BaF-LrI 1.0 4 Fraction survival p = 0.007 0.8 median survival + 2 days 3 Score # # 0.6 2 0.4 1 0.2 0 0.0 BaF BaF-LrI 0 5 10 15 20 Days after BaF injection Bindels et al, The ISME J 2016

Hypothetical role of the gut barrier Bindels & Thissen, Clin Nutr Exp 2015

Hypothetical role of the gut barrier Gut permeability BaF 1.5 CT (relative expression) BaF # # # mRNA levels BaF-LrI 1.0 * * * 0.5 ↘ Gut permeability ↗ 0.0 occludin ZO-1 Muc2 proglucagon Paneth cell differentiation and antimicrobials 1.5 (relative expression) ↘ Antimicrobial peptides ↗ # # mRNA levels 1.0 * # * * * 0.5 ↘ Immune system ↗ * 0.0  -defensins Reg3  TCF4 Lysozyme Pla2g2 2.0 * # (relative expression) # 1.5 mRNA levels # # # 1.0 ↘ Decreased in leukemic mice $ * * * 0.5 ↗ Increased by synbiotics * 0.0 CD3  Tbet IL-17A Foxp3 IL-10 Ebi3 Bindels et al, The ISME J 2016 lymphocytes

Current working model Gut barrier Cancer function cachexia Probiotics ? Propionate Prebiotics New metabolites ?

Outline 1. Gut microbiota as a nutritional target 2. Metabolic disorders associated with obesity 3. Metabolic disorders associated with cancer 4. Gut microbiota in alcohol-dependent patients

A role for the gut permeability? Leclercq et al, Brain Behav Immun 2012; Leclercq et al, Biol Psychiatry 2014.

A role for the gut permeability ? Leclercq et al, PNAS 2014

Dysbiosis Leclercq et al, PNAS 2014

Altered fecal metabolite profil Analysis of Volatile organic compounds by gas-chromatography-mass spectrometry (K. Verbeke, Kuleuven B) Bi-plot analysis reveals ADT1 HP- versus LP are differentiated (14 metabolites) Leclercq et al, PNAS 2014

Conclusions • Importance of the prebiotic concept. • Microbiota-dependent and independent effects of functional foods: strategies to demonstrate causality exist. • Underexplored areas could benefit from targeted prebiotic or synbiotic approaches.

UNL Gnotobiotic Carlos Hatem Rafael Mouse Facility Gomes Neto Kittana Segura Munoz Robert Schmaltz Brandon White Prof. Amanda Ramer-Tait FSR Fellowship Liz Cody Dr. Inés Martínez Maria Isabel Quintero Junyi Yang Prof. Jens Walter Maria Ximena Maldonado-Gomez

Post-doc position in July 2017 : laure.bindels@uclouvain.be Prof N. Delzenne Dr A. Neyrinck Prof P. Cani UCL (BE) ULG (BE) Rowett Institute, Aberdeen (UK) UCL (BE) Katholieke Universiteit Prof G. Muccioli Dr B. Taminiau Dr K.P. Scott and J.C. Martin Prof E. Hermans Leuven (BE) Prof G. Daube Prof P. Buc Calderon Dr B. Koener Dr H. Schoemans Prof J.P. Thissen Dr O. Schakman E. François Prof J. Maertens University of Reading (UK) Prof O. Feron Prof C. Blecker Prof S. P. Claus and C. Leroy Prof J. Mahillon Prof P. Sonveaux Prof J.B. Demoulin Prof A. Richel University of Alberta Dr P. Porporato Institut Pasteur, Lille (FR) Dr V. Havelange (CA) Prof B. Pot and Dr C. Grangette Dr J. Verrax Dr Fl. Bindels Prof J. Walter Dr R. Beck Dr I. Martinez