Download Genetics of Post Traumatic Stress Disorder and Other

Genetics of Post Traumatic
Stress Disorder and Other
Disorders in a
Multigenerational Family
Study of Traumatized
Armenians
UCLA
Julia N. Bailey, Ph.D
Armen Goenjian, M.D.
Ernie Noble, M.D., Ph.D.
Alan Steinberg, Ph.D.
Terry Ritchie, Ph.D.
Uma Dandert, Ph.D.
David Walling, Ph.D.
Armen Goenjian, M.D.
Haig A. Goenjian
Devon Schmidt
Ida S Karayan, Psy.
Armenia
Spitak Earthquake
LENINAKAN, DECEMBER 1988
LENINAKAN, 1988
Leninakan (Gumri), School No. 10
1990 Memorial. {743 out of 1010 had died}
Leninakan 1989
Mayor allocating land to build a
psychiatric clinic
PTSD
History of PTSD
The Anglo Saxon
Chronicle recounts a battle in
1003 A.D. between the
English and the Danes in
which the English commander
Alfred reportedly became so
violently ill that he began to
vomit and was not able to
lead his men.
Shell shock (Traumatic war
neurosis, neurasthenia)
• The Russians in the Russo-Japanese War
(1904-1905) were the first to diagnose mental
disease as a result of war stress
• During World War I shell shock was
considered a psychiatric illness resulting from
injury to the nerves
• The horrors of trench warfare meant that
about 10% of the fighting soldiers were killed
(compared to 4.5% during World War II)
Post Traumatic Stress Disorder
PTSD
A. Being exposed to a traumatic event
1. Experienced, witnessed or confronted
actual or threatened death or serious
injury
2. Felt Intense fear, helplessness or horror
Examples: War, hurricane, earthquake, rape,
beating, auto accident.
PTSD
B. The traumatic event is re-experienced
1. Recurrent intrusive thoughts, images
2. Distressing dreams
3. Act or feel as if the traumatic experience is
recurring experiences, illusions,
hallucinations , flashbacks
4. Psychological reactivity
. Hyper-arousal
5. Physical reactivity
1. Difficulty to sleep
2. Irritability
3. Difficulty to concentrate
4. Hyper-vigilance
Avoidance and Numbing
5. Startle response
1. Avoid thoughts, feelings
(Jumpiness)
2. Avoid people, places
3. Inability to recall aspects of the trauma
4. Diminished interest
5. Detachment, estrangement
6. Restricted range of affect
7. Sense of foreshortened future
D
C.
Prevalence
• Exposure to trauma
– Up to 90% in general population
• Lifetime rates of PTDS
– 1 in 9 for women
– 1 in 20 for men
PTSD Genetics
• Twin Studies
– Vietnam Era Twin (VET)
– Vancouver Twin Volunteers
• Family studies showed increase in PTSD,
anxiety and depression
Candidate Genes Studied
DiGangi et al. Biology of Mood & Anxiety Disorders 2013, 3:2
Systematic Gene Hunting
• No genome wide linkage scan
• GWAS (295 cases vs 195 controls)
– RORA retinoid-related orphan receptor alpha
gene.
– Replicated in 43 cases/41 controls African
American vets, not replicated in independent
100 cases/431 controls
Armenian
FAMILY STUDY OF PTSD
Psychiatric Outreach Program Staff,
Leninakan 1992
Ascertainment
• 12 nonrelated individuals from the four major
sections of Gumri, Armenia were invited to
participate in the study.
• Participants were acquaintances of the staff from
the Armenian Relief Society’s Psychiatric
Outreach Program.
• Chosen because they had large families willing
to participate.
Clinical Interview
Quantitative traits
• UCLA Post-traumatic Stress Disorder Reactive
Index (PTSD-RI)
• Beck Depressive Inventory (depression)
• Beck Anxiety Inventory (anxiety)
Dichotomous traits
• MINI (panic disorder)
• MINI (psychosis)
Earthquake-related profile
– Objective experiences
• including destruction of residence, deaths
of relatives, seeing dead bodies, being hurt,
and seeing someone else injured.
• Pre & post-earthquake traumatic
experiences.
– Subjective experiences
• fear of the earthquake, fear of getting badly
hurt or dying, and fear that someone else
will be badly hurt or die.
Subjects
• 212 participants from 12 multigenerational
families were recruited – 200 in study
– 7 of whom were not in Gumri during the earthquake
while 5 were unable to complete the testing.
• Families were 3-5 generations
• Mean and modal number of participants per
family was 16, range 13-19.
• 121 females, 79 males
Family 1
Family 2
Family 3
Family 4
Family 5
Family 7
Family 6
Family 9
Family 8
Family 10
Family 11
Family 12
Relative Pairs for 200 subjects
•
•
•
•
•
•
•
•
108 parent–offspring
137 full siblings
48 grandparent–grandchild
306 avuncular
64 grand avuncular
306 first cousins
225 first cousins once removed
111 second cousins.
ANALYSES & RESULTS
Experience During Earthquake
Objective
• 100% saw destroyed buildings throughout the city
• 90% witnessed dead bodies left lying in the streets
• 92% witnessed severely injured people.
Subjective experiences of fear during the earthquake
• 89% responded ‘a whole lot’ (very severe)
• 11% responded ‘a lot’ (severe)
Quantitative Genetic Variance
Component Model
phenotype of the j-th individual is assumed to be:
quantitative trait
loci (QTL)
effects
n
Additive polygenic effect
y j     q ji   x j  g j  e j
'
i 1
grand
trait
mean
covariates
environment
Significant Trauma Co-variates
Seen Death
Prior Trauma
Hurt
Secondary
Trauma
Panic
PTSD-RI
Sex
Anxiety
Age
Death of a Family
Member
Depression
Heritabilities
Goenjian et al. Psychiatric Genetics 2008, 18:261–266
Heritabilities of B,C,D PTSD
categories
Bailey et. al., Psychiatry Research 2010
Heritability
100%
80%
60%
Environm.
Genetic
40%
20%
66%
61%
Depress'n
Anxiety
41%
0%
PTSD
Goenjian, et. al. Psych. genetics, 2008; 18: 261 - 266
Bi-Variate Analyses
(Two Traits at once)
Phenotypic Correlation of PTSD
and Depression
Goenjian, Journal of Affective Disorders, 2012
Bi-Variate Analyses
Shared
environment
Vp
=
Vg
+
VH
Unique
environment
+
VE
2
  h21 h12 
  e21 e12 
 1121    g1 g12 
 
 I
 
  
 
2
2
2








 1222    g12 g 2 
 h12 h 2 
 e12 e 2 
the phenotype matrix
of covariance between
two traits
The genetic correlation indexes the similarity of
genes that contribute to variation in the two
traits and estimates the proportion of genes
shared in common between the two traits.
Bi-variate Correlations
Goenjian et al. Psychiatric Genetics 2008, 18:261–266
Candidate Genes
Candidate Genes Studied
to date
•
•
•
•
•
•
•
•
DRD2
DRD4
DAT
5HTP
TPH1
TPH2
COMT
DBH
(non) Association of DAT and
DRD2 with PTSD and subcategories
Bailey et. al., Psychiatry Research 2010
Association of TPH1, TPH2 and
SLC6A4 with PTSD symptoms
and depressive symptoms
Goenjian, Journal of Affective Disorders, 2012
Significant Associations
TPH1 –
TPH2SLC6A4
RS2108977 RS11178998
PTSD-RI
0.004
Depression 0.08
0.03
0.96
0.77
0.03
Goenjian, Journal of Affective Disorders, 2012
Future
DEEP SEQUENCING
(NEXT GENERATION)
Quicker & less expensive
Can now realistically sequence
individuals for their entire
genome.
Sanger Sequencing
Deep Sequencing
Overview of Deep Sequencing
• Deep sequence individual
– Just exoms
– Whole genome
– Whole genome under linkage peak
• Compare sequence to reference and identify
‘variants’.
– SNPS
– indels
– deletions
Variant Filtration Steps
(example)
1. Identify variants shared amongst affecteds in family but not with
family control*
2. Identify missense, nonsense, and splicing variants
3. Identify variants in conserved regions from 46-species alignment
4. Remove variants in 1000/5000 Genomes project and dbSNP
(excluding clinically associated SNPs)
5. Remove variants with SIFT score > 0.05
6. Remove variants with PolyPhen2 score <0.85
7. Compile a list of candidate genes based on disease model
* Can only be done in family studies, not case/control studies
Samples Selected for WES
• PTSD score (high for cases/ low for
control)
• Position in pedigree
• DNA amount
• Other affecteds in pedigree to test
candidate variants.
PTSD Armenian Family 7
Individuals chosen for Deep Sequencing
PTSD Armenian Family 4
Individuals chosen for Deep Sequencing
PTSD Armenian Family 12
Deep Sequencing Plans
•
•
•
•
Deep sequence 18 individuals
Sift for variants
Test candidates in other affecteds in family
Test candidates in other affecteds in other
families
• Test candidates in other Traumatized Armenian
Populations
• Test function
Summary
• PTSD is a complex, genetic trait that is difficult
to study due to exposure requirements,
difficulties of collecting samples in areas that
suffered severe trauma.
• Armenian family study is unique and may be
very useful in finding genes.