NuGO week 2016

Role of the gut microbiota in overand undernutrition Laure Bindels, PhD Copenhagen September 7, 2016

Adapted from Delzenne et al., Nat Rev Endocrinol 2011

Outline 1. 2. 3. 4.

Gut microbiota as a nutritional target Metabolic disorders associated with obesity Metabolic disorders associated with cancer 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 i.e. inulin-type fructans

Probiotics i.e. lactobacilli

Bacteriocins Antibiotics

FMT

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. 2. 3. 4.

Gut microbiota as a nutritional target Metabolic disorders associated with obesity Metabolic disorders associated with cancer 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

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

C26

… independent of the food intake Enterobacteriaceae (log10 [cells/g])

Daily food intake (% initial food intake)

*

11

125 100 75

#

CT (BaF) BaF CT (DR) DR

50 25

#

*

0

10

***

***

**

9 8 7 6

0

2

4

6

8

10

12

14

CT(BaF)

BaF

CT(DR)

DR

CT(BaF)

BaF

CT(DR)

DR

Days after BAF injection

Lactobacillus spp. (log10 [cells/g])

***

***

*** Parabacteroides goldsteinii ASF519 (log10 [AU/g])

* 10

9

8 CT(BaF)

BaF

CT(DR)

Bindels et al, The ISME J 2016

DR

11 10 9 8 7

1 .0

5.0 10 6

§ *

*

0 0

24

48

Acetate

72

mRNAlevels (relativeexpression)

Number of intact BaF3/w ell

BaF

B c r-A b l

control propionate 2mM

1.0 10 7

Propionate Butyrate

0 .5

0 .0

N D

CT BBaF3-ITF aF3

ITF

Acetate P r o p i o n a t e Propionate §Butyrate

6 0

µM

4 0

2 0

Acetate Propionate Butyrate

0

BaF3-ITF BaF3 CT

Bindels et al, Br J Cancer 2012; Bindels*, Neyrinck* et al, Plos ONE 2015.

Selected synbiotic approach D0

D1

D13

L. reuteri 100-23

Bcr-Abl-expressing BaF3 cells

Bindels et al, The ISME J 2016

†

ITF

With Bruno Pot & Corinne Grangette

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

0.45

1.0

# 0.5

Organ weight (% body weight)

mRNA levels (relative expression)

0.50

*

0.40 0.35 0.20

*

0.15

*$

0.10 0.05

ND 0.0

0.00

CT

CT

BaF BaF-LrI

BaF BaF-LrI CT

tibialis Morbidity score 5

1.2

4

1.0

3

## 2 1 0

BaF BaF-LrI

gastrocnemius

Survival

Fraction survival

Score

*#

BaF BaF-LrI p = 0.007 median survival + 2 days

0.8 0.6 0.4 0.2 0.0

BaF

BaF-LrI

0

5

10

15

Days after BaF injection

Bindels et al, The ISME J 2016

20

Hypothetical role of the gut barrier

Bindels & Thissen, Clin Nutr Exp 2015

Hypothetical role of the gut barrier Gut permeability

BaF mRNA levels (relative expression)

↘ Gut permeability ↗

1.5

#

#

1.0

*

*

CT BaF BaF-LrI

#

*

0.5

0.0

occludin

ZO-1

Muc2

proglucagon

Paneth cell differentiation and antimicrobials

↘ Antimicrobial peptides ↗

↘ Immune system ↗

mRNA levels (relative expression)

1.5

#

#

1.0

*

*#

*

*

0.5

* 0.0

Lysozyme -defensins Reg3

TCF4

Pla2g2

↘ Decreased in leukemic mice ↗ Increased by synbiotics

mRNA levels (relative expression)

2.0

*# #

1.5

0.5

*

$

*

*

*

0.0

CD3

Bindels et al, The ISME J 2016

#

#

#

1.0

Tbet

IL-17A

Foxp3

lymphocytes

IL-10

Ebi3

Current working model

Gut barrier function

Cancer cachexia

Probiotics ?

Propionate Prebiotics

New metabolites ?

Outline 1. 2. 3. 4.

Gut microbiota as a nutritional target Metabolic disorders associated with obesity Metabolic disorders associated with cancer 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.

Carlos Gomes Neto

Hatem Kittana

Rafael Segura Munoz

UNL Gnotobiotic Mouse Facility Robert Schmaltz Brandon White

Prof. Amanda Ramer-Tait

Liz Cody

Prof. Jens Walter

Dr. Inés Martínez

FSR Fellowship

Maria Isabel Quintero Junyi Yang Maria Ximena Maldonado-Gomez

Post-doc position in July 2017 : [email protected] Prof N. Delzenne Dr A. Neyrinck

Prof P. Cani

UCL (BE) Prof G. Muccioli Prof P. Buc Calderon Prof J.P. Thissen Prof O. Feron Prof P. Sonveaux Dr P. Porporato Dr J. Verrax Dr R. Beck

UCL (BE) Prof E. Hermans Dr B. Koener Dr O. Schakman Prof J. Mahillon Prof J.B. Demoulin Dr V. Havelange Dr Fl. Bindels

ULG (BE) Dr B. Taminiau Prof G. Daube E. François Prof C. Blecker Prof A. Richel

Katholieke Universiteit Leuven (BE) Dr H. Schoemans Prof J. Maertens University of Alberta (CA) Prof J. Walter Dr I. Martinez

Rowett Institute, Aberdeen (UK) Dr K.P. Scott and J.C. Martin University of Reading (UK) Prof S. P. Claus and C. Leroy Institut Pasteur, Lille (FR) Prof B. Pot and Dr C. Grangette