Gut Microbiome alterations in Major Depressive Disorder ... · 4 5 6 % Fibre in Diet in MDD & Beck...
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J. KELLY 1,3 , Y. BORRE 3 , S. EL AIDY 3 ,C. O' BRIEN 4 , J. DEANE 4 , P. KENNEDY 3 , G. MOLONEY 3 , L. SCOTT 1 , E. PATTERSON 4 , P. ROSS 4 , C. STANTON 4 , J. CRYAN 2,3 , G. CLARKE 1 , 3 T. DINAN 1,3 ; 1 Dept. of Psychiatry & Neurobehavioural Science, UCC 2 Dept. Anatomy & Neuroscience, UCC 3 Lab. of Neurogastroenterology, Microbiome Institute, Cork; 4 Teagasc Food Research Ctr., Cork 1. Dowlati, Y., et al., A meta-analysis of cytokines in major depression. Biol Psychiatry, 2010. 67(5): p. 446-57. 2. Lupien, S.J., et al., Effects of stress throughout the lifespan on the brain, behaviour and cognition. Nat Rev Neurosci, 2009. 10(6): p. 434-45. 3. Cryan, J.F. and T.G. Dinan, Mind-altering microorganisms: the impact of the gut microbiota on brain and behaviour. Nat Rev Neurosci, 2012. 13(22968153): p. 701-712. Methods The APC Microbiome institute is funded by Science Foundation Ireland (SFI). This publication has emanated from research conducted with the financial support of Science Foundation Ireland (SFI) under Grant Number SFI/12/RC/2273. The authors and their work were also supported by the Health Research Board (HRB) through Health Research Awards (grants no HRA_POR/2011/23; TGD, JFC and GC, HRA_POR/2012/32; JFC, TGD and HRA-POR-2-14-647: GC, TGD) and through EU GRANT 613979 (MYNEWGUT FP7-KBBE-2013-7). The Centre has conducted studies in collaboration with several companies including GSK, Pfizer, Wyeth and Mead Johnson. GC is supported by a NARSAD Young Investigator Grant from the Brain and Behaviour Research Foundation (Grant Number 20771). Conclusions Acknowledgements & Disclosure Email: [email protected] johnkelly@kellyjr83 UCCpsychiatry@UCCpsychiatry Introduction Aim Results • Alterations in the gut microbiota in patients with MDD are pronounced and may drive the prominent pathophysiological features of this disorder. The mechanisms underpinning these effects require further investigation. Ultimately, these findings may pave the way for therapeutic targeting of the gut microbiome as a viable strategy for novel antidepressant development. Gut Microbiome alterations in Major Depressive Disorder: Relevance to Pathophysiology • The biological mechanisms underlying the pathophysiology of MDD involve immune [1], endocrine [2] and neurotransmitter dysregulation. • Pre-clinical findings suggest that the gut microbiota can modulate brain development, function and behaviour by recruiting the same neuroimmune, neuroendocrine and neural pathways of the brain-gut-axis which are dysfunctional in MDD [3]. • However, the extent to which these pre-clinical findings translate to clinical populations is currently unknown. Determine the composition of the gut microbiota in patients with MDD compared to healthy control participants and its relationship to: • Short Chain Fatty acids (SCFAs) • Immune activity (plasma cytokines) • Hypothalamic-Pituitary-Adrenal axis (HPA-axis) function and • Tryptophan metabolism Demographics & Health Measures Healthy Controls (n=33) Depression (n=34) p-value Age mean (s.d.) 45.8 (11.9) 45.8 (11.5) 0.98 Sex Male (%) 19 (57.6) 21 (61.8) 0.73 BMI mean (s.d.) 24.58 (2.7) 26.2 (4.5) 0.07 Education, degree level (%) 26 (78.8) 8 (23.5) <0.001*** Relationship status (% yes) 26 (78.8) 8 (23.5) <0.001*** Employed (% yes) 31 (93.9) 16 (47.1) <0.001*** Smoker (% current) 3 (9.1) 13 (38.2) 0.003 Dyslipidaemia (%) 4 (12.1) 7 (20.6) 0.51 Hypertension (%) 3 (9.1) 3 (8.8) 0.97 IPAQ Low (%) 7 (21.2) 13 (39.4) 0.18 IPAQ Moderate (%) 16 (48.5) 14 (42.4) 0.62 IPAQ High (%) 10 (30.3) 6 (18.2) 0.26 Metabolic Equivalent Task Units (MET) median, range 1386 (7287) 693 (7722) 0.10 HAMD 17 median (range) NA 19.5 (14) NA Beck Depression mean, (s.d) NA 32.4 (9.92) NA Beck Anxiety median, (range) NA 25.5 (45) NA Duration of Depressive sx (months) median, (range) NA 3.0 (72) NA Number of Depressive episodes median (range) NA 1.0 (8) NA Depression in 1 st degree relative (%) 2 (6.1) 21 (61.8) <0.001*** Perceived Stress Scale (PSS) mean, (s.d) 7.5 (4.9) 27.7 (6.0) <0.001*** Pittsburgh Sleep Quality Index (PSQI) mean, (s.d) 2.8 (1.8) 11.7 (4.3) <0 .001*** Study Population: 34 patients with DSM IV MDD (MINI International Neuropsychiatric Interview) 33 healthy subjects matched for gender, age & ethnicity (see Table 1 for demographics & clinical characteristics) Measures: Gut Microbiota Structure & Diversity 16s rRNA gene sequencing Hypothalamic-Pituitary-Adrenal (HPA) Axis Salivary Cortisol (ELISA) Inflammatory Plasma Cytokines & C-Reactive Protein/CRP (Meso Scale Discovery) Tryptophan Metabolites Plasma tryptophan & kynurenine (HPLC) Subjective Mood & Stress Hamilton Depression rating scale (HAMD 17) Beck Depression & Anxiety scales (BDI & BAI) Perceived Stress scale (PSS) Pittsburgh Sleep Quality Index (PSQI) Diet & Exercise Food Frequency Questionnaire (FFQ) International Physical Activity Questionnaire (IPAQ) Kynurenine/Tryptophan Controls Depression 0.00 0.02 0.04 0.06 * Ratio CRP Controls Depression 0.0 0.5 1.0 1.5 2.0 2.5 3.0 ** ng/ml IL-6 Controls Depression 0.0 0.5 1.0 1.5 2.0 2.5 * pg/ml IL-2 Controls Depression 0.0 0.2 0.4 0.6 0.8 * pg/ml IL-8 Controls Depression 0 1 2 3 4 5 6 7 * pg/ml Cortisol Awakening Response 0 50 100 150 0 2 4 6 8 10 Depression Controls Timepoint (minutes from wakening) Cortisol concentration (nmol/l) Area under the Curve (AUCg) Controls Depression 0 200 400 600 800 1000 1200 * Cortisol (AUC G nmol/l) Death family member/friend Parental Upheaval Traumatic Sexual Experience Violence Illness/injured Other 0 10 20 30 40 50 Controls Depression * * Early Life Events (<17 years) a. * % Death family member/friend Relationship Upheaval Traumatic Sexual Experience Violence Ill/Injured Work related Other 0 10 20 30 40 50 Recent Life Events (last 3 years) b. ** * * % Psychotropic Medication in MDD group none SSRI only SNRI only TCA only MAOI only Bupropion only SSRI + NaSSA SNRI + NaSSA TCA + SARI SSRI + Bupropion SNRI + Bupropion Mood stabilizer Antipsychotic Benzodiazepine Z drug 0 5 10 15 20 25 30 35 40 c. % Altered Stress Response in MDD compared to Controls Fibre, Short Chain Fatty Acids (SCFAs) & Intestinal Permeability in MDD compared to Controls Inflammatory Profile in MDD compared to Controls Controls Depression 0 5000 10000 15000 Tryptophan ng/ml Controls Depression 0 200 400 600 800 Kynurenine ng/ml Controls Depression 0 2 4 6 Kynurenic Acid ng/ml Altered Gut Microbiota Composition, Diversity & Richness in MDD compared to Controls References Sample Characteristics Scan QR Code for link to UCC Psychiatry website poster and publication section Figure 1: Signaling pathways involved in The Brain Gut Microbiota Axis. Cryan & Dinan, Nat Rev Neuroscience, 2012 Figure 2: Brain Gut Microbiota Communication in Health & Disease. Grenham & Clarke et al., Frontiers in Physiology, 2011 Table 1: Comparisons between MDD patients & Healthy controls on demographics and clinical characteristics. Study participants were matched on the basis of age, BMI and ethnicity. Figure 2: a. Differences at Genus level between MDD and controls. b. Redundancy Analysis Plot differentiating MDD from Controls at the Genus level. c. The MDD group had significantly reduced richness p = 0.002, Chao1, p = 0.004 and Phylogenetic Diversity, p = 0.002. There was a trend reduction in the Shannon Index in the MDD group, p = 0.07. Percieved Stess Scale Controls Depression 0 10 20 30 40 Controls Depression *** Percieved Stress Scale % Fibre in Diet in MDD compared to Controls Controls Depression 0 1 2 3 4 % Fibre in Diet SCFAs in MDD compared to Controls Acetate Propionate Iso-Butyrate Butyrate 0 10 20 30 40 50 Depression Controls mMol/Kg 0 10 20 30 40 50 1 2 3 4 5 6 % Fibre in Diet in MDD & Beck Depression Scale Beck Depression Scale % Fiber in Diet Tryptophan Metabolite Profile in MDD compared to Controls Figure 3: Procedural stages of gut microbiota sampling & sequencing. Figure 5: Procedural stages of HPLC. Figure 4: Biomarker collection and analysis. Richness Controls Depression 0 200 400 600 ** Phylogenetic Diversity Controls Depression 0 10 20 30 40 ** Chao1 Controls Depression 0 200 400 600 ** Shannon Controls Depression 0 2 4 6 8 p = 0.07 Figure 1: a. Life events prior to the age of 17 years. The depressed group experienced significantly more parental upheaval (X 2 (1, N = 66) = 5.35, p = 0.02), traumatic sexual experiences X 2 (1, N = 66) = 4.24, p = 0.04, and violence X 2 (1, N = 66) = 4.70, p = 0.03 compared to the controls. b. Life events within the last 3 years. The depressed group experienced significantly more relationship upheaval X 2 (1, N = 66) = 7.76, p = 0.01, illness/injury X 2 (1, N = 66) = 4.24, p = 0.04. c. Psychotropic medication in the depressed group. a. b. c. Lipopolysaccharide binding protein Controls Depression 0 10 20 30 40 50 ng/ml a. b. c. d. Figure 3: a. There no significant differences in percentage fibre consumed between the groups as measured by a Food Frequency Questionnaire. b. There was a negative correlation between % Fibre consumed in diet and Beck Depression Scale in the MDD group. r = -0.43, p = 0.01. c. There were no significant differences in fecal SCFAs; acetate, propionate, iso-butyrate or butyrate between the groups. d. There were no significant differences in intestinal permeability as measured by Lipopolysaccharide binding protein. Figure 4: a. Salivary Cortisol Awakening Response. Repeated measures ANOVA across all sample collection time points revealed a significant main effect of group F (1,50) = 5.30, p=0.03). b. There was a significant increase in salivary cortisol as measured by Area under the Curve with respect to ground (AUCg) in the MDD group. p = 0.05. c. The MDD group had significantly higher PSS scores compared to the Control group. p = <0.001. a. b. c. Figure 5: a. The MDD had significantly higher levels of IL-6, p = 0.02, IL-2, p = 0.02, IL-8, p = 0.02 and CRP, p = 0.01 in the MDD group compared to the Control group. Figure 6: There was a significant difference in the tryptophan: Kynurenine ratio, p = 0.05. There were no significant differences in Tryptophan, p = 0.36, Kynurenine, p = 0.32 or Kynurenic Acid levels p = 0.70 between the groups.
Transcript of Gut Microbiome alterations in Major Depressive Disorder ... · 4 5 6 % Fibre in Diet in MDD & Beck...
J. KELLY1,3, Y. BORRE3, S. EL AIDY3,C. O' BRIEN4, J. DEANE4, P. KENNEDY3, G. MOLONEY3, L. SCOTT1, E. PATTERSON4, P. ROSS4, C. STANTON4, J. CRYAN2,3, G. CLARKE1,3 T. DINAN1,3; 1Dept. of Psychiatry & Neurobehavioural Science, UCC 2Dept. Anatomy & Neuroscience, UCC 3Lab. of Neurogastroenterology, Microbiome Institute, Cork; 4Teagasc Food Research Ctr., Cork
1. Dowlati, Y., et al., A meta-analysis of cytokines in major depression. Biol Psychiatry, 2010. 67(5): p. 446-57.2. Lupien, S.J., et al., Effects of stress throughout the lifespan on the brain, behaviour and cognition. Nat Rev Neurosci, 2009. 10(6): p. 434-45.3. Cryan, J.F. and T.G. Dinan, Mind-altering microorganisms: the impact of the gut microbiota on brain and behaviour. Nat Rev Neurosci, 2012. 13(22968153): p. 701-712.
Methods
The APC Microbiome institute is funded by Science Foundation Ireland (SFI). This publication has emanated from research conducted with the financial support of Science Foundation Ireland (SFI) under Grant Number SFI/12/RC/2273. The authors and their work were also supported by the Health Research Board (HRB) through Health Research Awards (grants no HRA_POR/2011/23; TGD, JFC and GC, HRA_POR/2012/32; JFC, TGD and HRA-POR-2-14-647: GC, TGD) and through EU GRANT 613979 (MYNEWGUT FP7-KBBE-2013-7). The Centre has
conducted studies in collaboration with several companies including GSK, Pfizer, Wyeth and Mead Johnson. GC is supported by a NARSAD Young Investigator Grant from the Brain and Behaviour Research Foundation (Grant Number 20771).
Conclusions
Acknowledgements & Disclosure
Email: [email protected]@kellyjr83UCCpsychiatry@UCCpsychiatry
Introduction
Aim
Results
• Alterations in the gut microbiota in patients with MDD are pronounced and may drive the prominent pathophysiological features of this disorder. The mechanisms underpinning these effects require further investigation. Ultimately, these findings may pave the way for therapeutic targeting of the gut microbiome as a viable strategy for novel antidepressant development.
Gut Microbiome alterations in Major Depressive Disorder: Relevance to Pathophysiology
• The biological mechanisms underlying the pathophysiology of MDD involve immune [1], endocrine [2] andneurotransmitter dysregulation.
• Pre-clinical findings suggest that the gut microbiota can modulate brain development, function andbehaviour by recruiting the same neuroimmune, neuroendocrine and neural pathways of the brain-gut-axiswhich are dysfunctional in MDD [3].
• However, the extent to which these pre-clinical findings translate to clinical populations is currentlyunknown.
Determine the composition of the gut microbiota in patients with MDD compared to healthy control participantsand its relationship to:
• Short Chain Fatty acids (SCFAs)• Immune activity (plasma cytokines)• Hypothalamic-Pituitary-Adrenal axis (HPA-axis) function and• Tryptophan metabolism
Demographics & Health Measures Healthy Controls (n=33) Depression (n=34) p-value
Age mean (s.d.) 45.8 (11.9) 45.8 (11.5) 0.98
Sex Male (%) 19 (57.6) 21 (61.8) 0.73
BMI mean (s.d.) 24.58 (2.7) 26.2 (4.5) 0.07
Education, degree level (%) 26 (78.8) 8 (23.5) <0.001***
Relationship status (% yes) 26 (78.8) 8 (23.5) <0.001***
Employed (% yes) 31 (93.9) 16 (47.1) <0.001***
Smoker (% current) 3 (9.1) 13 (38.2) 0.003
Dyslipidaemia (%) 4 (12.1) 7 (20.6) 0.51
Hypertension (%) 3 (9.1) 3 (8.8) 0.97
IPAQ Low (%) 7 (21.2) 13 (39.4) 0.18
IPAQ Moderate (%) 16 (48.5) 14 (42.4) 0.62
IPAQ High (%) 10 (30.3) 6 (18.2) 0.26
Metabolic Equivalent Task Units (MET) median, range 1386 (7287) 693 (7722) 0.10
HAMD 17 median (range) NA 19.5 (14) NA
Beck Depression mean, (s.d) NA 32.4 (9.92) NA
Beck Anxiety median, (range) NA 25.5 (45) NA
Duration of Depressive sx (months) median, (range) NA 3.0 (72) NA
Number of Depressive episodes median (range) NA 1.0 (8) NA
Depression in 1st degree relative (%) 2 (6.1) 21 (61.8) <0.001***
Perceived Stress Scale (PSS) mean, (s.d) 7.5 (4.9) 27.7 (6.0) <0.001***
Pittsburgh Sleep Quality Index (PSQI) mean, (s.d) 2.8 (1.8) 11.7 (4.3) <0 .001***
Study Population:34 patients with DSM IV MDD(MINI International Neuropsychiatric Interview)
33 healthy subjects matched for gender, age & ethnicity(see Table 1 for demographics & clinical characteristics)
Measures:Gut Microbiota Structure & Diversity16s rRNA gene sequencing
Hypothalamic-Pituitary-Adrenal (HPA) AxisSalivary Cortisol (ELISA)
InflammatoryPlasma Cytokines & C-Reactive Protein/CRP (MesoScale Discovery)
Tryptophan MetabolitesPlasma tryptophan & kynurenine (HPLC)
Subjective Mood & StressHamilton Depression rating scale (HAMD 17)Beck Depression & Anxiety scales (BDI & BAI)Perceived Stress scale (PSS)Pittsburgh Sleep Quality Index (PSQI)
Diet & ExerciseFood Frequency Questionnaire (FFQ)International Physical Activity Questionnaire (IPAQ)
Kynurenine/Tryptophan
Controls Depression0.00
0.02
0.04
0.06 *
Ra
tio
CRP
Controls Depression0.0
0.5
1.0
1.5
2.0
2.5
3.0 **
ng
/m
l
IL-6
Controls Depression0.0
0.5
1.0
1.5
2.0
2.5
*
pg
/m
l
IL-2
Controls Depression0.0
0.2
0.4
0.6
0.8
*
pg
/m
l
IL-8
Controls Depression0
1
2
3
4
5
6
7
*
pg
/m
l
Cortisol Awakening Response
0 50 100 1500
2
4
6
8
10Depression
Controls
Timepoint (minutes from wakening)
Co
rtis
ol c
on
cen
tra
tio
n (
nm
ol/
l)
Area under the Curve (AUCg)
Controls Depression0
200
400
600
800
1000
1200
*
Co
rtis
ol
(AU
CG
nm
ol/
l)
Death fa
mily
mem
ber/fri
end
Parenta
l Upheava
l
Traum
atic Se
xual E
xperie
nce
Viole
nce
Illness
/inju
red
Other
0
10
20
30
40
50
Controls
Depression
* *
Early Life Events (<17 years)a.
*
%
Death fa
mily
mem
ber/fri
end
Relatio
nship
Upheava
l
Traum
atic Se
xual E
xperie
nce
Viole
nce
Ill/In
jure
d
Work
rela
ted
Other
0
10
20
30
40
50
Recent Life Events (last 3 years)b.
**
*
*
%
Psychotropic Medication in MDD group
none
SSRI o
nly
SNRI o
nly
TCA only
MAOI o
nly
Bupropio
n only
SSRI +
NaSS
A
SNRI +
NaSSA
TCA + SA
RI
SSRI +
Bupro
pion
SNRI +
Bupro
pion
Mood st
abilize
r
Antipsy
chotic
Benzodia
zepin
e
Z dru
g
0
5
10
15
20
25
30
35
40
c.
%
Altered Stress Response in MDD compared to Controls
Fibre, Short Chain Fatty Acids (SCFAs) & Intestinal Permeability in MDD compared to Controls
Inflammatory Profile in MDD compared to Controls
Controls Depression0
5000
10000
15000
Tryptophan
ng
/ml
Controls Depression0
200
400
600
800
Kynurenine
ng
/ml
Controls Depression0
2
4
6
Kynurenic Acid
ng
/ml
Altered Gut Microbiota Composition, Diversity & Richness in MDD compared to Controls
References
Sample Characteristics
Scan QR Code for link to UCC
Psychiatry website poster and
publication section
Figure 1: Signaling pathways involved in The Brain Gut Microbiota Axis. Cryan & Dinan, Nat Rev Neuroscience, 2012
Figure 2: Brain Gut Microbiota Communication in Health & Disease. Grenham & Clarke et al., Frontiers in Physiology, 2011
Table 1: Comparisons between MDD patients & Healthy controls on demographics and clinical characteristics. Study participants were matched on the basis of age, BMI and ethnicity.
Figure 2: a. Differences at Genus level between MDD and controls. b. Redundancy Analysis Plot differentiating MDD from Controls at the Genus level. c. The MDD group had significantly reduced richness p = 0.002, Chao1, p = 0.004 and Phylogenetic Diversity, p = 0.002. There was a trend reduction in the Shannon Index in the MDD group, p = 0.07.
Percieved Stess Scale
Controls Depression0
10
20
30
40Controls
Depression***
Pe
rcie
ve
d S
tre
ss S
ca
le
% Fibre in Diet in MDD compared to Controls
Controls Depression0
1
2
3
4
% F
ibre
in
Die
t
SCFAs in MDD compared to Controls
Acetate Propionate Iso-Butyrate Butyrate0
10
20
30
40
50
Depression
Controls
mM
ol/
Kg
0 10 20 30 40 50
1
2
3
4
5
6
% Fibre in Diet in MDD & Beck Depression Scale
Beck Depression Scale
% F
ibe
r in
Die
t
Tryptophan Metabolite Profile in MDD compared to Controls
Figure 3: Procedural stages of gut microbiota sampling & sequencing.
Figure 5: Procedural stages of HPLC.
Figure 4: Biomarker collection and analysis.
Richness
Controls Depression0
200
400
600
**
Phylogenetic Diversity
Controls Depression0
10
20
30
40
**
Chao1
Controls Depression0
200
400
600
**
Shannon
Controls Depression0
2
4
6
8
p = 0.07
Figure 1: a. Life events prior to the age of 17 years. The depressed group experienced significantly more parental upheaval (X2 (1, N = 66) = 5.35, p = 0.02), traumatic sexual experiences X2 (1, N = 66) = 4.24, p = 0.04, and violence X2 (1, N = 66) = 4.70, p = 0.03 compared to the controls. b. Life events within the last 3 years. The depressed group experienced significantly more relationship upheaval X2 (1, N = 66) = 7.76, p = 0.01, illness/injury X2 (1, N = 66) = 4.24, p = 0.04. c. Psychotropic medication in the depressed group.
a. b.
c.
Lipopolysaccharide binding protein
Controls Depression0
10
20
30
40
50
ng
/m
l
a. b. c. d.
Figure 3: a. There no significant differences in percentage fibre consumed between the groups as measured by a Food Frequency Questionnaire. b. There was a negative correlation between % Fibre consumed in diet and Beck Depression Scale in the MDD group. r = -0.43, p = 0.01. c. There were no significant differences in fecal SCFAs; acetate, propionate, iso-butyrate or butyrate between the groups. d. There were no significant differences in intestinal permeability as measured by Lipopolysaccharide binding protein.
Figure 4: a. Salivary Cortisol Awakening Response. Repeated measures ANOVA across all sample collection time points revealed a significant main effect of group F (1,50) = 5.30, p=0.03). b. There was a significant increase in salivary cortisol as measured by Area under the Curve with respect to ground (AUCg) in the MDD group. p = 0.05. c. The MDD group had significantly higher PSS scores compared to the Control group. p = <0.001.
a. b. c.
Figure 5: a. The MDD had significantly higher levels of IL-6, p = 0.02, IL-2, p = 0.02, IL-8, p = 0.02 and CRP, p = 0.01 in the MDD group compared to the Control group.
Figure 6: There was a significant difference in the tryptophan: Kynurenine ratio, p = 0.05. There were no significant differences in Tryptophan, p = 0.36, Kynurenine, p = 0.32 or Kynurenic Acid levels p = 0.70 between the groups.
