Moshe Szyf, PhD Presentation at 2016 Science of HOPE The workshop will discuss basic epigenetic mechanisms and their role in setting up gene expression programs as well as review evidence from animal and human studies illustrating how epigenetic processes might be mediating the impact of the social and physical environment on the mental and physical health of children as well as adults. The implications of epigenetics for developing social policies, new interventions, diagnostics and therapeutics will be presented.

1. M M M MM
M
Epigenetic mechanisms mediating between
experience and mental and physical health
Moshe Szyf
Department of Pharmacology and Therapeutics
McGill University, Montreal Canada
The Science of Hope
Foundation for healthy generations Seattle, Washington,
April 19 2016

2. CH3
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DNA METHYLATION REACTIONS

3. Epigenetic marks (i.e. DNA methylation) at critical positions silence
genes;
same gene could be either active or silent based on the state of methylation
X
Tet H2OH
X

4. DNA methylation inhibits RNA Pol2
complex binding
.
TBP
TBP

5. CH3
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methylase
demethylase
active
inactive
The gene is an equilibrium between activity and lack of activity that is defined by
epigenetic marks. Epigenetic marks program how genes work in different organs
in our body;
This equilibrium could be reset by environmental signals
nutrients
toxins
social
behavioral pathological
physiological

6. DNA METHYLATION a mechanism of adaptation of the genome
in response to changing environments without changing the
sequence of DNA
•Evolutionary time scale
•Trans-generational time scale
•Life long time scale
•Life cycle stations time scale
•Seasonal time scale
•Physiological proximal time scale

7. Bioenvironment Physical environment Social environment
phenotype
Bioenvironment Physical environment Social environment
Health challenges
CH3
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CH3
System wide
CH3
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Signaling pathways
Adaptive response of the genome in early life

8. LOW LG HIGH LG
STRESS RESPONSE STRESS RESPONSE
A bar in Madrid, licking rats and the birth of the field of “behavioral epigenetics”
Pups
Adults

9. Biological mother Adoptive mother
Low LG
Low LGHigh LG
High LG
Behavioral programming by the mother is epigenetic not genetic;
The fostering mother and not the biological mothers transfers the phenotype
Cross-fostering /adoption
experiment

10. PKA
Behavioral gene programming is stable but nevertheless reversible
5-HT
cAMP
NGFI-A
HAT
Demethylase
MBD2?
DNMT
TSA
Methionine

11. The signature of maternal care in non-
human primates
Mother Surrogate
Stephen Suomi

12. Genome wide view of the differential methylation between
maternal and nursery reared rhesus monkeys in T cells and
prefrontal cortex

13. PROMOTER METHYLATION RESPONSE TO early life stress Maternal deprivation in
T CELLS AND PFC
Rearing = mother
/ nursery
high
low
methylation
PFC T cells

14. Quebec ice storm 1998
Storm32 score
Threat
Loss
Scope
Change

15. DNA methylation signatures of randomized objective stress during pregnancy in
human adolescent T cells Quebec ice storm of 1998
storm32
449
30
1206
879
709
948
1368
135
287
51
1210
384
39
8
1191
1248
204
826
9
115
1365
827
1391
1157
405
1316
1315
56
1427
1060
105
1030
1036
363
100CpGsites
storm32 heatmap
storm32
storm32 heatmap
stress%
low%
medium%
high%
high%
low%
methyla1on%
The$DNA$methylation$signature$of$maternal$stress$in$chord$blood$T$cells:

16. Correlation between objective randomized stress during pregnancy and CG methylation
in the SCG5 gene in T cells of 15 year old
Objective stress during ice storm 1998
Methylation

17. Ice storm data suggested enrichment of type 1 diabetes pathway
Type 1 diabetes signaling pathway enriched with differentially
methylated genes that correlate with objective stress

18. DNA methylation mediates the long term impact of
adult experience, for example, chronic pain, addiction;
Diagnostic and therapeutic implications

19. Transcription
factor
XCH3 CH3
X
Transcription
factor
X
AcAc Ac
Transcription
Machinery
Transcription
machinery
DNMT
Demethylases
Protein A Protein A
X
Phenotype A Phenotype B
RG108
SAM
Behavioral intervention

20. Transcription
factor
XCH3 CH3
X
Transcription
factor
X
AcAc Ac
Transcription
Machinery
Transcription
machinery
DNMT
Demethylases
Protein A Protein A
X
Phenotype A Phenotype B
Figure 1
RG108
SAM
Behavioral intervention

21. Incubation model of cocaine craving
Copyright 2013, Western Washington University,
Behavioral Neuroscience Program/Grimm lab
RG108, SAM
or aCSF
injections
Days
cocaine (Coc.) or saline (Sal.)
self-administration (SA)
withdrawal
10 days
0 1 Day 30
Day 1 groups:
- Sal. SA: Sal.
- Coc. SA no cue: 1W
- Coc. SA cue: 1C
Day 30 groups:
- Coc. SA no cue: 30W
- Coc. SA cue: 30C
+
- Coc. SA cue-aCSF: aCSF
- Coc. SA cue-RG108 cue: RG108
- Coc. SA cue-SAM: SAM
- Coc. SA cue-PPT
- Coc. SA cue-roscovitine
RG108 or aCSF
injections
Days
10 days
0-2cocaine
self-administration
a
b
FIGURE 1
Day 60
withdrawal
Day 60 groups:
- Coc. SA cue-aCSF: aCSF-60
- Coc. SA cue-RG108 cue: RG108-60
- Coc. SA cue-SAM: SAM-60
EPM, locomotion,
memory tests
Withdrawal intensifies craving
Yadid

22. DNA methylation changes following cocaine
administration, cues and withdrawal

23. Epigenetic modulation reprograms craving behavior only at
the end of a long withdrawal period when paired with a cue
Combined behavioral and epigenetic therapy
b
0
500
1000
1500
0
50
100
150
200
250
Training Test
0
20
40
60
80
aCSF
RG108
Distance(cm)
Timeinopen
arm(sec)
caCSF
RG108
aCSF
RG108
Exploration(%)
d
0
20
40
60
80
** *
0
30
60
90
120 ***
*
a
Leverpresses
D30 D60
D30 D60
aCSF
SAM
Leverpresses
f
aCSF
RG108
0
30
60
90
120
e
Leverpresses
Before injection
After injection
*
*
**
b
0
500
1000
1500
0
50
100
150
200
250
Distance(cm)
Timeinopen
arm(sec)
caCSF
RG108
0
30
60
90
120 ***
*
D30 D60
aCSF
SAM
Leverpresses
f
aCSF
RG108
0
30
60
90
120
e
Leverpresses
Before injection
After injection
b
FIGURE 5
0
500
1000
1500
0
50
100
150
200
250
Training Test
0
20
40
60
80
aCSF
RG108
Distance(cm)
Timeinopen
arm(sec)
caCSF
RG108
aCSF
RG108
Exploration(%)d
0
20
40
60
80
** *
0
30
60
90
120 ***
*
a
Leverpresses
D30 D60
D30 D60
a
S
Leverpresses
f
aCSF
RG1
0
30
60
90
120
e
Leverpresses
Before injection
After injection
*
*
**
RG108 DNA methyl transferase inhibitor
After long withdrawal immediately after training
SAM DNA methylation stimulator

24. ESR1 and CDK5 are modulated by DNA methylation inhibitors in the cocaine craving model; ESR1 agonist propyl
pyrazole triol (PPT) and CDK5 antagonist (roscovitine) reverse cocaine craving
Esr1mRNA
0.0
0.4
0.8
1.2 *
aCSF roscovitine
CDK5 inhibitor
0
400
800
1200
0
30
60
90
120
150
**
Leverpresses
Locomotion
0.0
0.4
0.8
1.2
1.6
aCSF PPT
ESR1 agonist
0
400
800
1200
0
20
40
60
80
100
Leverpresses
****
Locomotion
Cdk5mRNA
FIGURE 7
Cdk5
chr4
0
-0.78
500 b
Esr1
27 kb
chr1
0
-0.39
Esr1mRNA
0.0
0.4
0.8
1.2 *
aCSF ros
CDK5 inhib
4
8
12
0
30
60
90
120
150
**
Leverpresses
Locomotion
0.0
0.4
0.8
1.2
1.6
aCSF
ESR1 agon
4
8
12
0
20
40
60
80
100
Leverpresses
****
Locomotion
Cdk5mRNA
aCSF RG108
DNMT inhibitor
aCSF RG108
DNMT inhibitor
DNAmethylationDNAmethylation
a
b

25. What are the implications of a life-long dynamic
epigenome?
chemical social
Signaling pathways
phenotype
biosphere
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26. Acknowledgements
Anne McKiney
Tony Huang
Ziv Machnes
Gustavo Turecki
Michael Meaney
Ian Weaver
Patrick McGowan
Michael Hallett,
Matt Suderman
Stephen Suomi, Allyson Bennett, Peter Pierre
Nadine Provencal
Renaud Massart
Laura Stone
Lei Cao
Suzan King
Gal Yadid (BIU)
Renaud Massart
Royi Barnea (BIU)
Yahav Dikshtein (BIU)
Oren Meir (BIU)
Peter Gass
Michaela Schmidt
THIS RESEARCH IS SUPPORTED BY
GRANTS FROM CIHR MRC, the Sackler program in
epigenetics and psychobiology