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Microbial metabolite production
by gut microbes and implications
for host health
Catherine Stanton
APC Microbiome Institute (APC) & Teagasc Food
Research Centre, Moorepark, Fermoy, Co. Cork,
Ireland.
E-mail: [email protected]
Microbiome is defining in terms of human health
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Human
~10 trillion cells
Bacterial Community
~23 thousand genes
…a hidden organ
Microbiota
~100 trillion cells
~3 million genes
Intestinal Microbiota
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Complex and dynamic community
1014 microorganisms/g contents
10 times more bacteria than human cells
100-fold more unique genes than our own genome
Important functions:
METABOLIC
MORPHOLOGIC
PROTECTIVE
Guarner et al., 2011
• Fermentation of food components
• Production of SCFAs, vitamins, bioactives
• Control of epithelial cell proliferation
• Development of the immune system
• Barrier effect
• Production of antimicrobial substances/pathogen
inhibition
Microbiota Impacts Host Metabolism
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e.g. SCFA
Ryan & Delzenne, 2016.
Microbiome has intimate ties in almost every facet of host health
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5
4
2
1
3
Gut Microbes and Obesity
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Felix Sommer & Fredrik Bäckhed
Nature Reviews Microbiology 11, 227-238 (April 2013)
Gut Microbes & Diabetes
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Brain-Gut-Microbiota Axis
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Human Gut Microbiota
-large potential for health
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Bioactive
metabolites
PERTURBED MICROBIOTA
• Necrotizing enterocolitis
• Obesity
• Diabetes
• Metabolic syndrome
• Irritable bowel syndrome
• Inflammatory bowel disease
•Colon cancer
• Altered CNS development
GI microbiota over lifetime
Your microbiota assembles in the first years of life
and remains relatively stable until old age
Assembly
Stability
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Collapse
Opportunities for interventions/diagnostics, for food, biotech & pharma
APC Hypothesis
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We can mine
the gut
microbiota for
products that
promote
health or treat
disease
•
•
•
•
•
Diagnostics
Antibiotics
Anti-inflammatories
Medical foods
Functional foods
Mining the Gut Microbiota for Bioactives
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Pharmabiotics
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The gut
microbiota
plays a role in
determining
health - we can
mine for
pharmabiotics
•
•
•
•
•
•
Faecal transplants
Microbial consortia
Probiotics
Prebiotics
Metabolites
Bacteriophage
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Microbiota
Immune System
Modulation of Intestinal Microbiota
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“Open window of opportunity”
(Fouhy et al., 2012)
Importance of the correct establishment of intestinal microbiota
Early postnatal period  constitutes key moment
Modulation to the establish a healthy microbial profile
Early life: Setting
the la
immune
balance forinmune
life
Edad temprana:
Base de
homeostasis
para toda la vida
-9 months
-9 meses
Nacimiento
Birth
~ 36 meses
~ 36 months
Alergia
Allergy
Obesidad
Obesity
Enfermedades
Coronary
Heart
coronarias
Diabetes
Diseases
Enfermedades
Diabetes
autoinmunes
Autoimmunity
Salud
HEALTHY
LIFE
Bajo riesgo
de
enfermedad
↓ Risk
disease
Kerperiem et al., 2012
Mode of Delivery: Phylum level
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Hill et al (2017) (MICROBIOME)
Increase in Diversity over Time
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Hill et al (2017) (MICROBIOME)
By Year 2 Microbiota Becomes More Adult-like
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C)
A)
100%
Unclassified
d = 0.05
Other
90%
Clostridium_IV
Coprococcus
80%
23.9%
Alistipes
Erysipelotrichaceae incertae sedis
70%
Clostridium sensu stricto
Gemmiger
60%
Clostridium XI
4.7%
Prevotella
50%
B)
Clostridium XVIII
Ruminococcus
Shannon Diversity
40%
Clostridium XlVa
Roseburia
30%
Anaerostipes
20%
Lachnospiracea incertae sedis
Blautia
10%
Faecalibacterium
Bacteroides
Year 1
Year 2
***
0%
Year 1
Year 2
***
Bifidobacterium
(data unpublished)
Pharmabiotics
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Conjugated
Isomers
Bacteriocins
SCFA
Gut
Bacterial
Exopolysaccharides
Metabolites
Vitamin B
Bioactive
Peptides
Neuroactive
Compounds
Antihypertensive
Antithrombitic
Caseinophosphopeptides
Immunomodulatory
Antioxidant
Opioid
Cytomodulatory
Conjugated Linoleic Acid (CLA)
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Anti-Cancer
Improves
Heart Health
Improves
Immune
System
Regulating
Improves Body
Composition
Bone
Formation
Fat
Recombinant Lb. paracasei producing t10, c12
CLA in vivo
GLC
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LA
Lactoacillus paracasei
t10, c12 CLA
Expressing CLA isomerase (P. acnes)
Increased t10, c12 CLA content of adipose tissue following dietary intervention
0.03
g/100 g FAME
***
0.02
n=8
0.01
8 weeks
0.00
Vector
Control
CLA
Producing
Rosberg-Cody et al (2011) Microbiology
Production of Bioactive Lipids - CLA
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90
80
Bifidobacterium breve
70
60
LA
50
%
49
Conversion
to CLA 30
20
10
GLC
CLA
Bifidobacterium longum
Linoleic acid (g/100g FAME)
Administered B. breve is metabolically active in
the gut
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Linoleic acid + B. breve
Linoleic acid
r= -0.863
c9, t11 CLA (g/100g FAME)
Wall et al., AJCN 2009 89: 1393-1401
Impact of administered B. breve on FA composition
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***
LA + B. breve
LA + B. breve
LA
Control
*
*
BALB/c mice
SCID mice
Wall et al., AJCN 2009, 2012
Pigs
Elevated Lc-PUFA in Adipose Tissue and Brain
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EPA (Eicosapentaenoic Acid)
DHA (Docosahexaenoic Acid)
Unsupplemented
*
***
B. breve 702258 supplemented
n=8
* = p<0.05
*** = p<0.001
Wall et al. Am. J. Clin. Nutr. 2009
Microbiome-mediated therapies for heart health?
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Probiotic
• BSH
• EPS
Prebiotic
• Arabinoxylan
• Oat β-glucan
Archaebiotics
• TMADepletion
EPS Producing LAB: Probiotic for Heart Health?
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EPS
Lb. mucosae 6426: Genome Sequence
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The genome revealed a plethora of
homologues linked to carbohydrate
metabolism (EPS operon) and mucin
Lactobacillus
mucosae DPC6426
binding. In addition, several putative
bile salt hydrolase genes (BSH) were
identified, along with a potential novel
phage and CRISPR elements.
DPC6426 Reduces Atherosclerosis – In Vivo
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High Fat Control
Heat Killed
n = 14
n = 14
n = 14
A 24-week intervention with either Live (lDPC) or Heat-Killed (hkDPC) Lb. mucosae DPC6426 fed in
conjunction with a high-fat diet significantly attenuated cholesterol (and in particular LDL-C) accumulation
when compared to the high-fat control (HFC). In turn, the strain reduced the development of aortic plaque
in the atherosclerosis-prone apolipoprotein-E-deficient (apo-E-/-) mouse model.
Bile Salt Hydrolase Activity & Specificity
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Porcine
Murine

(Ryan et al., unpublished)
Live Lactobacillus mucosae
Alters Microbiome Composition & Functionality
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hkDPC
(Ryan et al., unpublished)
HFC
16S MiSeq
U/w UniFrac

liveDPC
Faecal 1H-NMR OPLS-DA
liveDPC
HFC
*
hkDPC
*
*
*
Antiatherogenic Activity of DPC6426
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Brain-gut-microbiota axis
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•
Brain gut axis is a bi-directional
signalling between the brain and the
GI tract (O’Mahony 2011; Dinan and
Cryan, 2016)
• Gut microbiota may play a role in
neuronal development and plasticity,
modulating pain perception and even
behaviour (Forsythe et al., 2010; Dina
et al., Biol Psychiatry. 2013)
•
Microbes produce a wide spectrum of
neuroactive compounds, including fatt
acids, GABA (Lyte, 2011; Barrett et
al., 2012)
•
Studies have shown the ability of
probiotics to alleviate distress in
humans and an anxiolytic-like activity
in rats (Lyte, 2011)
Microbes and neurotransmitters
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Genus
Neurochemical
Lactobacillus, Bifidobacterium
GABA
Streptococcus, Escherichia, Enterococcus,
Lactococcus, Lactobacillus
Serotonin
Escherichia, Bacillus
Escherichia, Bacillus, Lactococcus, Lactobacillus,
Streptococcus
Lactobacillus, Bacillus
Norepinephrine
Dopamine
Lactobacillus, Lactococcus, Streptococcus,
Enterococcus
Histamine
Acetylcholine
From Wall et al., 2014; Bacterial Neuroactive Compounds Produced by Psychobiotics
Adv Exp Med & Biol 817, pp 221-241
GABA = Microbial Metabolite
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• γ-aminobutyric acid (GABA) is synthesised through
decarboxylation of glutamic acid by glutamic acid
decarboxylase (GAD)
• GABA: main inhibitory neurotransmitter in the CNSbiological functions of GABA are mediated by activation
of GABA receptors in the brain and periphery
91 intestinally-derived bacteria assessed, one
Lactobacillus brevis strain (Lb. brevis DPC 6108)
could convert up to 90% MSG to GABA in vitro
Anti-anxiety effect of GABA producing
Lb brevis 6108
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Unsupplemented
GABA powder
L. brevis DPC 6108
Inner Zone duration
Inner Zone distance moved
0.0511
50
600
500
*
400
cm
30
300
20
200
10
100
0
0
Total distance moved
5000
4000
*P<0.05
cm
sec
40
3000
2000
1000
0
(Marques et al., unpublished)
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Sunday 28th June 2015
Gut Microbes, Metabolites and Health
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• Gut Microbiome-central role in host metabolism
• Microbial metabolism-range of signalling molecules,
which directly and indirectly impact on host lipid
metabolism/cholesterol synthesis/brain function
• Probiotics have potential, but understanding
mechanisms critical
• Future studies required to fully comprehend role of
gut microbiome in: host-lipid metabolism, CNS
development, ageing
• Targeted gut microbiome modulation dietary
interventions for optimised health throughout life
Acknowledgments
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COLLABORATORS
Paul Ross
Paul O’Toole
Ger Fitzgerald
Fergus Shanahan
Ted Dinan
John Cryan
Noel Caplice
Eamonn Quigley
Tony Ryan
Gene Dempsey
RESEARCHERS
Paul Ryan
Rebecca Wall
Elaine Patterson
Ruairi
Robertson
Lis London
Tatiana Marques
Mairead Coakley
Eva Rosberg
Alan Hennessy
Eoin Barrett