How life events get under the skin: Implications for suicide - - PowerPoint PPT Presentation

How life events get “under the skin”: Implications for suicide prevention

Gustavo Turecki MD PhD

William Dawson Chair Douglas Hospital Research Centre McGill University

Turecki et al Trends in Neurosciences 2012

CSA, CPA and Suicide Attempts

Fergusson et al. Child Abuse & Neglect, 2008

5 10 15 20 25

Suicide attempts at age 16-18 according to CSA and CPA types

None Contact Penetration None Regular Severe

CSA CPA

Suicide and Childhood Adversity

Suicide completers

41.4 29.3 6.8 5 10 15 20 25 30 35 40 45 Any abuse Suicides Psychiatric controls Community Controls % Ostovar et al, 2009

Abuser identity and suicide risk

Brezo et al, British J Psychiatry 2008

HOW CAN EVENTS OCCURRING DURING CHILDHOOD INCREASE LIFETIME SUICIDE RISK?

The effect of age and experience on children’s ability to discriminate 2 non-English contrasts

Werker & Tees, 2005

Sensitive periods in musical training

Difference in anterior corpus callosum volume according to age of musical training Penhune, 2011

Glucocorticoid Receptor (GR) regulation, quality of maternal care, and cognitive function in rats

Regulation of hippocampal GR expression by maternal care…. … affects behavioral phenotype in adult rats.

Francis et al., Science 1999; Caldji et al., PNAS 1998

Ecker, Nature 2012

Gene

Transcription Factors RNA polymerase II

DNA chr chromosome

• mosome

Gene ene 1 Gene ene 2

DNA CYTOSINE METHYLATION

C mC

SAM-CH3 SAM DNMT

H3C

Environmental influences

Environmental influences modify epigenetic marks around our genes and modify their expression

Promoter Promoter

CH3 CH3 CH3 CH3 CH3 CH3

Turner and Muller, Mol. Endo. 2005

The GR Gene: complex promoter in rats and humans

GR promoter determine in which tissue GR is expressed. Exon 1-7 (in rat) and 1F (in human) are specific to the hippocampus. Exon 1-7 and its homologue 1-F posses binding sites for the transcription factor NGFIA.

NGFIA transcription factor binding site

Weaver et al., Nat:Neurosci 2004

GR methylation, GR expression and cognitive function in rats

Nerve growth factor-inducible protein A (NGFI-A)

HPA Axis

HPA and suicide risk

N = 78 inpatients DST = 1mg DST

Coryell & Schlesser, AJP 2001

Risk = 26.8%

Risk = 2.9% 1990-2008 – primary health settings N = 786,868 oral glucocorticoid treatment courses N = 372,696 patients

Fardet et al, AJP 2012

MGSS Brain Bank

– Ongoing brain bank collection

• Suicides and sudden-death controls without medical

intervention

– Psychological autopsies

• Semi-structured interviews with an average of two

informants per case

• Clinical vignette
• Assessment by a blind panel for best-consensus

DSM IV diagnoses

• Validation with medical records

www.douglasrecherche.qc.ca/suicide

GR expression is decreased in the hippocampus

• f abused suicide completers

McGowan et al, Nature Neurosci 2009

Site-specific hypermethylation in hGR1F promoter in abused suicide completers

NGFI-A transcription factor’s binding site

McGowan et al, Nature Neuroscience 2009

Human

15 Maternal gestational stress Gestational and birth-related complications History of childhood adversity Psychiatric diagnosis

Studies:

• 1. Hompes et al. 2013
• 2. Hogg et al. 2013
• 3. Oberlander et al. 2008
• 4. Radtke et al. 20 1

1

• 5. Mulligan et al. 2012
• 6. Filiberto et al. 20 1

1

• 7. Bromer et al. 2012
• 8. El Hajj et al. 2013
• 9. McGowan et al. 2009

1

b

Footnotes:

• a. CpG 12 & 13with fear of delivery all trimesters
• b. CpG 6 with fear of delivery
• c. site- and parameter-specific:

CpG 38/39 (near NGFI- A site) ↓ with fear of changes T1 & 2; CpG 36 ↑ with fear of integrity T1 & 2; CpG 36 ↑ with fear of delivery T3

• d. CpG 1-3, NGFI- A

site

• e. CpG 1 & 3 only (NGFI- A

site is CpG 3 & 4)

• f. CpG 35 (NGFI- A

site is CpG 37 & 38)

• g. CpG 1 and 5
• 10. Perroud et al. 20 1

1 1

• 1. T

yrka et al. 2012

• 12. Melas et al. 2013
• 13. Perroud et al. 2013
• 14. Dammann et al. 20 1

1

• 15. Labonte et al. 2012

1

c

, 3

d

, 4, 5 6, 7 9, 10, 1 1

e

, 12

f

, 13 14

g

1

c

Direction of Findings

8 1

a

2

NR3C1 First Exon V ariants

Cortisol Amygdala Hippocampus Hypothalamus

Normal development Normal levels of GR in the hippocampus Exposure to early –life adversity Low levels of GR in the hippocampus Stress Stress activates HPA axis Stress activates HPA axis Secretion of cortisol Secretion of cortisol Binding of cortisol to GR in the hippocampus Binding of cortisol to GR in the hippocampus (poor binding) Lack of inhibition of the HPA axis

Overactive HPA axis

Inhibition of the HPA axis (negative feed back) GR

Adrenal cortex Anterior Pituitary CRH AVP ACTH

Comparative methylation patterns associated with early environment across species

Suderman et al, PNAS 2012

Genome-wide study Immunoprecipitation (MeDIP)-chip

Weber et al, Nature Genet 2005

N abused suicide: 27 N controls: 17

Differentially methylated sequences and chromosomal distribution

Hypermethylated Hypomethylated

Chromosome Hypermethylated Hypomethylated

N = 110 31% N = 245 69%

N = 355

P = 0.67

Inverse correlation between whole-genome expression and promoter methylation r = -0.14, p≤4.6x10-247

Labonté et al, Archives of General Psychiatry, 2012

Category Enrichment Score: 2,106 Count % P-value MF protein homodimerization activity 13 4,39 0,0054 MF protein dimerization activity 17 5,74 0,0093 MF identical protein binding 19 6,42 0,0095 Category Enrichment Score: 1,683 Count % PValue BP negative regulation of immune response 4 1,35 0,0062 BP negative regulation of immune system process 6 2,03 0,0085 BP negative regulation of response to stimulus 6 2,03 0,0180 BP regulation of immune effector process 4 1,35 0,1975 Category Enrichment Score: 1,625 Count % PValue BP cell-cell adhesion 11 3,72 0,0095 BP cell adhesion 18 6,08 0,0373 BP biological adhesion 18 6,08 0,0378 Category Enrichment Score: 1,545 Count % PValue CCs cell soma 8 2,70 0,0091 CCs cell projection 18 6,08 0,0174 CCs neuron projection 11 3,72 0,0215 CCs dendrite 5 1,69 0,1926 Category Enrichment Score: 1,481 Count % PValue BP positive regulation of biosynthetic process 21 7,09 0,0044 BP positive reg. of macromolecule biosynthetic process 19 6,42 0,0106 MF transcription activator activity 14 4,73 0,0106 BP positive reg. of macromolecule metabolic process 23 7,77 0,0107 BP positive reg. of nitrogen compound metabolic process 18 6,08 0,0186 BP positive regulation of transcription 16 5,41 0,0248 MF transcription factor activity 24 8,11 0,0254 BP positive r. of transcription from RNA pol. II promoter 12 4,05 0,0258 BP positive reg. of nucleobase, nucleoside, nucleotide and nucleic acid metabolic process 17 5,74 0,0280 BP positive regulation of transcription, DNA-dependent 14 4,73 0,0299 BP positive regulation of gene expression 16 5,41 0,0311 BP positive regulation of cellular biosynthetic process 18 6,08 0,0313 BP positive regulation of RNA metabolic process 14 4,73 0,0316 MF transcription regulator activity 33 11,15 0,0361 MF sequence-specific DNA binding 16 5,41 0,0466 BP regulation of transcription 49 16,55 0,0768 BP regulation of transcription from RNA pol. II promoter 17 5,74 0,0860 BP regulation of transcription, DNA-dependent 35 11,82 0,0863 BP transcription 40 13,51 0,0999 BP regulation of RNA metabolic process 35 11,82 0,1087 MF DNA binding 42 14,19 0,1861

Most significantly enriched functional annotation clusters

Protein dimerization Immune response Cell adhesion Cell/neuronal plasticity Gene-expression regulation

MAO IGR

MAOA DMRs Methylation levels 7150 bp after MAOA 3’ end 12300 bp after MAOB 3’ end

MAOA: The “warrior” gene

FITC-A

A B

Non Neuronal Fraction (56.1%) Neuronal Fraction (32.7%)

D C F E

SSC-A

1500 1000 500

Cell Count

100K 200K

102 103 104 105 102 103 104 105

Fluorescence-assisted cell sorting

20 40 60 80 100 1 2 3 4 5 6 7 8 9 10 11 12- 13

% of Methylation

CpGs

% of Methylation in MAOA IGR Non Neuronal Fraction (+)

Suicide CTRL 20 40 60 80 100 1 2 3 4 5 6 7 8 9 10 11 12-13

% of Methylation

CpGs

% of Methylation in MAOA IGR Neuronal Fraction (+)

Suicide CTRL 15 30 45 60 75

% of Methylation Total % of Methylation in MAOA IGR Non Neuronal Fraction (+)

Suicide CTRL 30 45 60 75 90

% of Methylation Total % of Methylation in MAOA IGR Neuronal Fraction (+)

Suicide CTRL

* * * *

MAO findings

MAOB MAOA

100% 0%

*

Hypomethylation in suicide completers Hypomethylation in the neuronal cell fraction Low Exp. in Suicide

*

Low levels of H3K27me3 in suicide

*

Suicide CTRL

High Exp. in Suicide

*

8 kb

• lincRNA in MAOA/B intergenic region
• Repressed by epigenetic mechanisms
• DNA methylation and chromatin states

change in suicides

• Increased lincRNA expression
• lincRNA regulates MAOA expression in the brain

LincRNA

Ulitsky & Bartel, Cell 2013

• RNAs longer than 200bp
• No open reading frame
• Poorly conserved

evolutionarily

• Highly expressed in the CNS
• Play a role in neurogenesis

Viral-Mediated Gene Transfer

IGR

Technical Validation

• Cannulation
• Transfection (GFP)

Expression

• MAOA/MAOB
• lincRNA

Behavioral Assessment

• Depression/Anxiety
• Impulsivity
• Aggression

HSV HSV

Intra DG lincRNA HSV Infection

Pre Screening Agressive behavior PT1 PT2 PT3 PI1 PI2 PI3 PI4 PI5 PI6 PI7 PI8 Re Sreening Agressive behavior PI0

Sacrificing and collecting tissue

Peak lincRNA expression

20 40 60 80 100 day1 day2 day3 % Animal Attacking

% Animal Attacks

GRIP Prevalence Cohort

• longitudinal school cohort representative of the Quebec general

popluation

• 1509 boys & 1509 girls
• multiple informants

Wave I Childhood Ages: 6-12

Assessment frequency: yearly Informants: teachers & parents Variables : childhood personality, family socioeconomic circumstances

Wave II Adolescence ages: 15-18

Assessment frequency:

• nce

Informants: adolescents & parents Variables: psychiatric history & suicidality

Wave III Adulthood ages: 19-24

Assessment frequency:

• nce

Informants: young adults Variables: psychopathology, suicidality, social support, partner violence, childhood abuse, etc

Wave IV Adulthood ages: mid-20s Informants: subjects

• DNA
• Attempt Hx
• Personality variants
• Psychopathology

Suicide Attempts Childhood Adversity 2.11 (1.51-2.97) Ages 6 – 12 years Age 21 years

Childhood Adversity and Risk of Suicide Attempts

P<0.001

Wanner et al, Psych Medicine 2012

Life Trajectories

Brezo et al, Arch Pediatr Adolesc Med. 2008

Adjusted risk = 60% increase in likelihood of suicide attempts

A Presence of externalizing disorders: Full mediation

Nagelkerke R2=.14 OR=1.60a (1.19-2.15) OR=2.43c (1.49-3.97) OR=2.07b (1.29-3.31)

B Absence of externalizing disorders: Partial mediation

OR=2.07b (1.29-3.31)

a.01<P<.05, b.001<=P<=.01, cP<.001

Nagelkerke R2=.14 OR=11.52c (2.14-61.91)

Childhood Adversity Suicide Attempts High Anxiousness Childhood Adversity Suicide Attempts High Anxiousness

Anxiousness mediates the effect of childhood adversity on suicide attempts differently according to the presence of disruptive disorders

Wanner et al, Psych Medicine 2012

Impaired decision making in SA: The Iowa Gambling Test

Jollant et al, AJP 2005

Delis-Kaplan Executive Function Test:

TSST effect on Word Color Inhibition (Stroop Effect) and Trail Making Test

Group*TSST = F(1,25)=5.51, p<.05 McGirr et al, JPN 2010 Group*TSST = F(1,25)=6.13, p<.05

0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 0.5 Drug Dep Drug Abuse Alcohol Dep Alcohol Ab SCZ MD Bipolar Anxiety Dis Suicides Controls

6-month Axis I Prevalence rates in Suicides and Controls from Montreal

N = 197 Kim et al, Psych Med 2003

Summary

Increased suicide risk

Early life adversity

CH3 CH3 CH3 CH3 CH3 CH3 CH3 CH3 CH3

Stable gene function regulation ( mRNA)

Genes involved in the regulation of behavior (i.e. GR)

Behavioral and emotional trait regulation

Anxiety  Impulsivity

Chronic substance use Poor decision making

Impaired regulation

• f emotional and

cognitive responses Faciliation of acts

Depressive psychopathology Suicidal ideation

Turecki et al Trends in Neurosciences 2012

Acknowledgements

• MGSS

– Naguib Mechawar – Volodymyr Yerko – Benoit Labonté – Gilles Maussion – Jennie Yang – Marie-Martine Beaulieu – Gang Chen – Carl Ernst

• McGill University

– Moshe Szyf – Michael Meaney – Michael Hallett – Matt Suderman

• University of Montreal

– Richard Tremblay – Frank Vitaro – Brigitte Wanner

• Johns Hopkins

– Robert Casero

• University of Texas

– Howard Gershenfeld

• Cleveland Clinic

– Veronique Lefebvre

• Support: CIHR, NIMH,

FRSQ, AFSP, NARSAD

• Thanks to the Families