Shackman Psyc210 Module11 NatureNurture Part2 032415

We
wish to suggest a structure for the salt of

deoxyribonucleic acid...This structure has novel
features which are of considerable biological
interest...It has not escaped our no;ce that the
specic pairing we have postulated
immediately suggests a possible copying
mechanism for the gene;c material

Watson & Crick Nature 1953

we should remain unabashed about the ul;mate

[goal] of genomic medicine, which [is] to transform
the health of our children and our childrens children

Lander Nature 2011

Plan for Today

Lecture on Nature & Nurture, Part 2
The genomics revolu;on
Part of the mo;va;on is to educate you
as ci;zens, taxpayers, and
healthcare consumers
Take-home cri;cal thinking ques;ons
PSYC 210:
The nature & nurture of T&P, Part 2

Molecular substrates of trait-like
dierences with a focus on
Gene-Environment Interac@ons

AJ Shackman
March 2015
Conceptual Roadmap
T&P are somewhat heritable

What molecular mechanisms account for that
heritability?

T&P reects the inuence of both gene;c inheritance
(nature) and the environment/experience (nurture)

Are these separate eects (G and E) or an
interac@on (G x E)?

e.g., Exposure to stress and risky genes
Conceptual Roadmap

heritability?


Are these separate eects (G and E) or an
interac@on (G x E)?

e.g., Exposure to stress and risky genes
Conceptual Roadmap

heritability?


Are these separate and independent eects
(G and E) or an interac@on (G x E)?

e.g., Risky genes AND stress exposure
DNA
Quick Gene;cs Primer
Genes
DNA Chromosomes
- DNA is organized into chromosomes, the vectors of heredity
- Human cells have 23 pairs of chromosomes (46 / cell), one pair
descended from mom and one from dad
DNA Chromosomes
- DNA is organized into chromosomes, the vectors of heredity
- Human cells have 23 pairs of chromosomes (46 / cell), one pair
descended from mom and one from dad
What about genes?
hQps://en.wikipedia.org/wiki/Human_genome
Genes
- Gene: a region of DNA/RNA sequence, corresponding to a unit of

inheritance or single basic instrucZon (recipe)
- Human genome: ~21,000-23,000 genes
- Genes hold the informaZon to build and maintain an organism's
cells and pass geneZc traits to ospring
- Genes are transcribed to RNA and used
to code protein synthesis, e.g., build
neurons, axons, transporters, vesicles,
neurochemicals, myelin, etc.

DNA/Gene messenger RNA protein
Genes

- Human genome: ~22,000 protein-coding genes

Genes


Genes


What makes us dierent?
Alleles = Individual Dierences

- Allele: a variant of a gene (geneZc polymorphism)

- Dierent alleles can result in dierent observable phenotypic traits,
such as dierent hair color
- Most genes do not dier across individuals; only a single allele
- When people talk about good or bad genes, they actually mean
allele (individual dierences in geneZc instrucZons)


such as dierent hair color (cf. Weasley clan)




SNPs = Individual Dierences

- SNP: single nucleoZde polymorphism (so, a parZcular kind of allele)
- 10-30 million SNPs in humans (~10M common [>1% frequency])
- Most common type of sequence
variaZon (90% total variaZon)
SNPs = Individual Dierences

- SNP: single nucleoZde polymorphism (so, a parZcular kind of allele)
- ~20 million SNPs in humans (~10M common [>1% frequency])
- Most common type of sequence
variaZon (90% total variaZon)
How might we try to discover the

alleles that predict trait-like
dierences in T&P (N, E, C)
or intermediate phenotypes (e.g.,
amygdala ac@va@on)?
Lets start with a

simplied example
PaZents (Cases)
Allele #1
Allele #2
% Allele #1
90%
hQp://vassarstats.net/odds2x2.html
PaZents (Cases)
Controls
Allele #1
Allele #2
% Allele #1
90%
10%
hQp://vassarstats.net/odds2x2.html
PaZents (Cases)
Controls
Allele #1
Allele #2
% Allele #1
90%
10%
Allele #1 is associated with a 9x

increased risk of disease!
(risk ra;o)
hQp://vassarstats.net/odds2x2.html (0.90 allele #1 = paZents; 0.10 allele #2 = paZent; 0.90/0.10 = 9 )
Now we need to scale this up
GWAS
- GWAS: genome-wide associaZon study (repeat for >1M common
SNPs)

- Example: Individuals with the G-allele of SNP1 (rs1333049) were
overrepresented amongst pa;ents (upper row) vs. controls
GWAS
- GWAS: genome-wide associaZon study (repeat for >1M common
SNPs)

SNP1
Pa;ents
Controls
SNP2
Etc.
How the heck are we going to

eciently measure >1M SNPs?
SNP Chip
Details Are Not Important
SNP Chip
SNP Chip
Fragment of one strand
SNP Chip
Binds with complement
SNP Chip
Array of probes for

dierent gene variants
SNP Chip
Measure binding
Array of probes for
SNP Chip
Measure binding
Array of probes for
GWAS
- Brute force approachtesZng one-by one the correlaZon between
traits and hundreds of thousands of common geneZc variants
- GWAS treats every common genomic variant the same, allowing the
discovery of novel trait- or disease relevant geneZc variants in the
absence a priori hypotheses
- The opposite of candidate gene studies that uZlize theories to test
a small number of geneZc variants
- Penalty is low staZsZcal power and sensiZvity, due to correcZon for
millions of tests across the genome
- Furthermore, GWAS will miss rare geneZc variants because they are
not included on the SNP chip arrays
GWAS
GWAS
GWAS
millions of tests across the genome (p<.05 p<.05/1,000,000)
GWAS
millions of tests across the genome (p<.05 p<.05/1,000,000)
not included on the chip
Why bother?
The Premise
We are precy ignorant about the biology linking genotypes to phenotypes (e.g., N, E, and SC). So its
dicult to generate good hypotheses (candidate genes to examine).

If we could somehow harness gene;cs to understand the molecular neurobiology of T&P and associated
psychiatric disorders, it would enable us to:

-
Redene diagnosZc categories and T&P traits in terms of quanZable eZology (root causes)
Develop novel treatments or prevenZon eorts targeZng links in the eZological chain
IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)
Precision medicine: Predict treatment response or more quickly pick the best treatment (e.g., carriers of a
parZcular polymorphism)
Enhance prognosis: You have 3 months to live
Monitor treatment in terms of changes in the underlying neural systems
Provide a novel discovery tool for addressing some of the most fundamental quesZon about the nature of T&P
- Where does extraversion come from?
-
What is the biological basis of childhood shyness?
How are T&P, on the one hand, and psychopathology, on the other, related?
The Premise


-
Precision medicine: Predict treatment response or more quickly pick the best treatment (e.g., carriers of a
parZcular polymorphism)
Enhance prognosis: You have 3 months to live
Monitor treatment in terms of changes in the underlying neural systems
-
The Premise


-
Predict treatment response or more quickly pick the best treatment (e.g., carriers of a parZcular
polymorphism).personalized or precision medicine
Enhance prognosis: You (and other carriers of SNP X) have 3 months to live
-
The Premise


-
-
The Premise


-
-
The Premise


-
-
The Premise


-
-
The Premise


-
-
Some Success with GWAS

Very recent GWASs have iden;ed sets of polymorphisms that
collec;vely account for much of the heritability of some major
psychiatric disorders

e.g., 32% varia;on in MDD (phenotype) can be explained by
individual polymorphisms (genes)
- Suggests that a substanZal proporZon of geneZc variaZon results
from very large numbers of small eect variants

Very recent GWASs have iden;ed sets of gene;c polymorphisms
that collec;vely account for much of the heritability of some major

- In general, recent work suggests that most geneZc variaZon results




But theres a poten@al problem
The Problem of Small Eects

- Common polymorphisms have, at most, weak eects on brain
funcZon and behavior (e.g., 0-5%)
- Small eects are hard to detect and likely to result in
nonreplicaZons (false negaZves)
- Prompted the development of large-scale consorZums and data-
sharing networksthousands of subjects across dozens of labs
provides the staZsZcal power needed to reliably detect weak eects
- But this also begs the quesZon of so what why bother if the main
eect of individual geneZc polymorphisms is so small

nonreplicaZons (false negaZves) (Study 1 hit, Study 2 null eect,
Study 3 hit, etc.)
sharing networksthousands of subjects across dozens of labs
provides the staZsZcal power needed to reliably detect weak eects
- But this also begs the quesZon of so what why bother if the main
eect of individual geneZc polymorphisms is so small

nonreplicaZons (false negaZves) (Study 1 hit, Study 2 null eect,
Study 3 hit, etc.)
sharing networkstens of thousands of subjects across dozens of
labs provide the staZsZcal power needed to reliably detect weak
eects

Whadya mean small eects?
The Case of Height
The Case of Height

Height is highly heritable (~85%) and highly polygenic, with many
variants contribuZng and therefore small eect sizes for the individual
polymorphisms

Using discovery and validaZon samples of ~130,000 and ~50,000
subjects, Lango Allen et al. discovered 180 variants

AssociaZons were on the order of 14mm (0.02 0.2% of phenotypic
variaZon; ~70 mm or 7 cm)
The Case of Height

Height is highly heritable (~85%) and highly polygenic, with many
variants contribuZng and therefore small eect sizes for the individual
polymorphisms

Using discovery and validaZon samples of ~130,000 and ~50,000
subjects, Lango Allen et al. discovered 180 variants

AssociaZons were on the order of 14mm (0.02 0.2% of phenotypic
variaZon)
The Case of Height

To put 4 mm (per variant) in perspec@ve
Tony Stark (5.7 feet)

Vs.
Black Widow (5.25 feet)

= 15 cm = 150 mm

= 37.5 polymorphisms

Millimeters? What about good old

fashioned inches?
1 inch = 25.4 mm =

6.4 allelic variants
What about T&P?
In contrast to height and psychopathology, GWAS studies of T&P have relied on much
smaller samples
De Moor used a discovery sample of discovery samples of N = 17, 375
Service had a discovery sample of N = 11,000
Not surprisingly, they lacked the power to detect a single geneZc variant that was
signicantly associated with dierences in N/NE or E/PE L

ElucidaZon of geneZc loci signicantly inuencing temperament and personality will
require potenZally very large samples, and/or a more rened phenotype.
smaller samples
De Moor et al.: N = 17, 375 Service et al.: N = 11,000
signicantly associated with dierences in N/NE or E/PE L

smaller samples
signicantly associated with dierences in N/NE or E/PE a|er correcZng for
mulZple comparisonsL

smaller samples
signicantly associated with dierences in N/NE or E/PE a|er correcZng for
mulZple comparisonsL

Are small eects a big problem?

Is the expenditure worth it?
Tiny Eects Can Be

Prac@cally Important

Meyer et al Amer Psychol 2001
Tiny Eects Can Be


Meyer et al Amer Psychol 2001
Tiny Eects Can Be

The Case of Asprin

Rosnow
At a special meeZng held December 18, 1987, it was decided to end prematurely a randomized
double blind experiment (n=22l) on the eects of aspirin on reducing heart aQacks. The reason
for the unusual terminaZon of this experiment was that it had become so clear that aspirin
prevented heart aQacks (and deaths from heart aQacks) that it would be unethical to conZnue
to give half the subjects a placebo. Now what do you suppose was the magnitude of the
experimental eect that was so dramaZc as to call for the terminaZon of this research?

Actually, [it was] r = .034.

Greenwald
This was a signicant (P<0.00001) reducZon [from 2.16% to 1.27%] in the risk of heart aQack
among those in the aspirin group. Applying the studys esZmated risk reducZon of 44% to the
2010 U.S. Census esZmate of about 46 million male U.S. residents 50 or older, regular small
doses of aspirin should prevent approximately 420,000 heart aQacks during a 5-year period

Rosnow Amer Psychol 1990; Greenwald et al JPSP in press; but see also Ferguson Rev Gen Psychol 2009
Tiny Eects Can Be

The Case of Asprin

Rosnow


Greenwald

Tiny Eects Can Be

The Case of Asprin

Rosnow


Greenwald

Tiny Eects Can Be

Take Home Point

Small eects can be very prac;cally important when they are cumulated over many

Individuals (e.g., the popula;on of na;ons)
Individual experiences (e.g., many job applica;ons, in the case of discrimina;on)
What about more direct evidence?
The Case of Cholesterol:

Small Eects, Big Therapeu@c Impact
Lander Nature 2011; PapassoZropoulos TiCS 2015

~1 in 3 deaths
CDC 2010 Data


Cholesterol is a strong predictor of heart disease risk

N = 900,000 subjects; Lancet 2007



Cholesterol is a strong predictor of heart disease risk

1 mmol/L lower total cholesterol = 50%
reducZon in mortality at mid-life

Cholesterol gums up your arteries

N = 900,000 subjects; Lancet 2007

Heart Disease Risk is Inherited
Cholesterol is about 50% heritable
Molecular pathways?

GWAS (N > 100,000)

IdenZed 95 signicant SNPs
Together accounZng for 10% of the phenotypic variance and 25% of
the geneZc (heritable) variance
In short, each SNP only accounted for about 0.1% of the variance in
observed cholesterol levels
Sounds awful, huh?

Molecular pathways?

GWAS (N > 100,000)

Sounds awful, huh?

Molecular pathways?

GWAS (N > 100,000)

Sounds awful, huh?

Molecular pathways?

GWAS (N > 100,000)

Sounds awful, huh?

One of the 95 loci = HMGCR
HMGCR predicts modest variance in cholesterol
2.8 mg/dl
But the pathway coded by this locus is the target of the
StaZns, the most popular drug for reducing cholesterol
Among the best selling drug in history ($12.4B in 05)
Clinically eecZve: Lower LDL cholesterol by
~1.8 mmol/l, which translates into an esZmated 60% decrease in the
number of cardiac events (heart aQack, sudden cardiac death)


One of the 95 loci = HMGCR
HMGCR predicts modest variance in cholesterol
2.8 mg/dl
But the protein coded by this locus is the target of the
StaZns, the most popular drug for reducing cholesterol
Among the best selling drug in history ($12.4B in 05)
Clinically eecZve: Lower LDL cholesterol by
~1.8 mmol/l, which translates into an esZmated 60% decrease in the
number of cardiac events (heart aQack, sudden cardiac death)


Take Home Points
We just discovered 94 other SNPs
Lots of opportuniZes for understanding the molecular
mechanisms underlying one of the biggest killers
More generally small eects in a GWAS do not necessitate small
impact

Now you might be saying to yourself,

Im a scal conserva@ve, what is this
cos@ng me and other taxpayers?

Are these gene@c discoveries
worth it?

The Cost of GWASs: Is it Worth It??
- If we assume that the GWAS results (described in our review) reect
a total of 500,000 SNP chips @ $500/chip = $250M
- ~2,000 variants detected in high-quality biomedical studies (across a
variety of physical and mental disorders)
- $125,000 per discovered variant
- Good investment?
- $250M
- 1-2 stealth ghter jets
- Much less than a single navy submarine
- FracZon of the ~$9 billion cost of the Large Hadron Collider
- Equivalent to ~100 5-year NIH grants to individual labs
Visscher et al 2012

- Been esZmated that all the high-quality GWAS studies (circa 2012)
reect a total of 500,000 SNP chips @ $500/chip = $250M
- Detected ~2,000 geneZc variants across a variety of physical and
mental disorders
- $125,000 per newly discovered variant
- Good investment?
- $250M
- Equivalent to ~100 5-year NIH grants (R01s) to individual labs
Visscher et al 2012

mental disorders
- Good investment?
- $250M
Visscher et al 2012

mental disorders
- Good investment? Judgment call
- $250M
Visscher et al 2012

mental disorders
- Good investment? Judgment call
- $250M
Visscher et al 2012
Perhaps a dierent
strategy would be helpful?
Main Eects and Interac;ons

- TradiZonal GWAS approaches assume that
individual genes directly determine phenotypes
(T&P, disorders)
But what if traits and disorders reect

the interac@on of Genes and
Experience?

What if we inherit a predisposi@on to
be neuro@c, extroverted, or
conscien@ous, given the appropriate
environment?
G-E Interac;ons (G x E)
- Traits cluster in families (remember Draco Malfoy)
- But heritability appears to be probabilisZc and parZcular family
members may not inherit a parZcular trait (e.g., high levels of
evilness, self-control, or depression)
- Neither geneZcs nor environment is solely responsible for
producing trait-like dierences in phenotypes

- Individuals can inherit sensiZvity to the eects of various
environmental and experienZal factors
- Same pathogen, dierent outcomes
e.g, sunlight exposure has a much stronger inuence

on skin cancer risk in the fair-skinned

environmental and experienZal factors


environmental and experienZal factors (diathesis)


environmental and experienZal factors (diathesis)

Can apply this logic to gene hun@ng
Search for combina@ons of genes (those contribu@ng to skin pigmenta@on) and

environments (exposure to sunlight) that predict outcomes of interest
G*E Illustra;ve Examples

What kind of environmental factors are we talking about?

- Family Conict: Individuals who are geneZcally predisposed)to low C/SC are even more impulsive in a
conictual family environment;
-
Marriage and Religiosity: Individuals who are geneZcally predisposed to substance abuse are less likely to
develop drinking problems if they were married or religious; Gene*Marriage also found for MDD
Low Parental Monitoring and Substance-Abusing Peers: Individuals who are geneZcally predisposed to
substance use and anZsocial behavior are more likely to develop problems in these environments
In short, a wide variety of environmental factors can

(a) trigger,
(b) compensate for, or

(c) enhance geneZc predisposiZon


-

(a) trigger,



-

(a) trigger,



-

(a) trigger,



-

(a) trigger,

(c) enhance
par;cular gene;c predisposi;ons
What about molecular gene@cs (SNPs)?
A Famous Illustra;on
monoamine oxidase A (MAOA) gene
Caspi et al Science 2002
Do genes interact with maltreatment to predict an@social behavior?

Do alleles change the slope rela@ng maltreatment to AS behavior?
Yes! Abused kids with the good allele (gray) showed fewer
an@social behaviors protec@ve
Second Famous G*E Example

Caspi (Duke)
Caspi et al Science 2003, Amer J Psychiatry 2010; Monroe Psychol Sci 2008

Caspi (Duke)

Caspi (Duke)
Does the impact of stress on depression depend

on the serotonin transporter (5-HTT) gene?

Caspi (Duke)
Short
Long
Yes! Carriers of the Short allele showed a strong posi@ve

rela@onship between life stress and depression Short allele
confers risk (or the Long allele confers protec@on)
Key Take Home Points
Key Take Home Points for Todays Mee;ng

1. T&P are somewhat heritable
2. We can use GWAS (SNP chips) to search the genome (10M common variants)
for the SNP's that account for that heritability
3. The premise of this genomic approach is nothing short of revoluZonary. We
remain largely ignorant about the biological underpinnings of T&P. GWAS
provide a powerful tool for discovering the molecular origins of T&P
Redene T&P and associated disorders in terms of root causes instead of
self-reported symptoms
Develop novel screening procedures and intervenZons grounded in an
understanding of underlying mechanisms (eZology)
4. Brute force approach. Penalty is low staZsZcal power due to correcZon for
millions of tests(p<.05 vs. p<.05/1,000,000).
5. Heritable traits reect the inuence of many SNPs (polygenic), each with a very
small eect.
6. Not surprisingly, preliminary GWASs for T&P (N<20,000) have failed to detect
signicant genes. Need huge samples.

small eect.

small eect.

small eect.

small eect.

small eect.

7. Small eects
Are challenging and expensive to detect
Can be pracZcally and clinically/therapeuZcally important (e.g., baseball,
smoking, ibuprofen)
Do not necessarily imply mechanisZcally worthless (HMGCR/StaZns
example)
8. Yes, GWAS is expensive, but not as expensive as many other big Zcket items
funded by tax payers
9. O|en the impact of the environment (sunlight) on outcomes (cancer) is
dependent on genes (pigmentaZon). Individuals can inherit sensiZvity/diathesis.
10. In some cases, the importance of a geneZc variant will only be unmasked by
examining the G-E interacZon (there is no main eect of the gene on the
outcome of interest).


7. Small eects
Can be pracZcally important (e.g., baseball, smoking, ibuprofen)
example)


7. Small eects
example)
G-E interacZon.


7. Small eects
example)
G-E interacZon.

To be con@nued
Genome
Intermediate Phenotype
Traits (Evildoing)
Cri;cal Thinking Take Home Ques;ons
Con@nued

1.
Pick any one of the fundamental quesZons about

T&P that we have covered in class since the start of
the semester (e.g., what are the fundamental
dimensions of T&P) and then briey discuss how
geneZc informaZon could prove helpful for
addressing that quesZon
2.
It appears likely that genomics will play an

increasing role in medicine, and hence is likely to
have a direct impact on you and your loved ones
(directly, as parZcipants in a public healthcare
system, or as taxpayers funding R&D). IdenZfy one
way in which the genomics revoluZon could
inuence you (e.g., screening, precision medicine)
and discuss whether this is a posiZve or negaZve
thing. You may nd it helpful to spend a liQle Zme
on Wikipedia (hQps://en.wikipedia.org/wiki/
IntroducZon_to_geneZcs)
Con@nued

1.

dimensions of T&P). Briey discuss how gene;c
informa;on could prove helpful for addressing
that ques;on
2.

Con@nued

1.

that ques;on
2.

Con@nued

1.

that ques;on
2.

system, or as taxpayers funding R&D). Iden;fy one
way in which the genomics revolu;on could
inuence you and discuss whether this is a
posi;ve or nega;ve thing. You may nd it helpful
to spend a liQle Zme on Wikipedia (hQps://
en.wikipedia.org/wiki/IntroducZon_to_geneZcs)
Con@nued

3. In March 2015, researchers at Weill Cornell showed "that a single-nucleoZde polymorphism (SNP) in the
faQy acid amide hydrolase (FAAH) gene of the endocannabinoid system has parallel molecular, neural and
behavioral eects in both humans and mice engineered to express the variant human allele" (Dincheva
Nature Comm 2015)

Available @ hQp://www.nature.com/arZcles/ncomms7395.epdf?
referrer_access_token=mm5H5NOSq1Jk_TvYe8eEzdRgN0jAjWel9jnR3ZoTv0MFY2u7ZdbRZ8yhzdTOJfa-
Q9HKMjv_5rGSvPOZGVRItjvxxd3D6Fuj0xbHtGY8oldf7UYS6IrqEzfzB5TJv1BbzdIXz-
nmzLYfNCGxS84JQNwH25BbPI31H6kc8SFO8TU1kO2mN4-
psU-996CDETtbNnR_CIJf88376uvUOsssvlA2IkZzxf_viisyTH6B8pyD2QRPa1RJvpWrvVaNnjH&tracking_referr
er=blogs.scienZcamerican.com

They note that, "Non-replicaZon of candidate gene associaZon studies has been a major problem within
the eld of behavioral geneZcs. Here, we have used a parallel mousehuman experimental approach that
enables greater control for environmental and geneZc confounds. The convergent ndings reported here
establish that eects of geneZc variaZon in FAAH are evident at neural and behavioral levels in both the
mouse and in humans, providing a persuasive cross-species validaZon."

Con@nued

Nature Comm 2015)



Con@nued

Nature Comm 2015)



Con@nued

The report was highlighted in a provocaZve essay in the NY Times enZtled The Feel-Good Gene (6 March
2015)

Available @ hQp://www.nyZmes.com/2015/03/08/opinion/sunday/the-feel-good-gene.html?_r=0

The author notes that

"For the rst Zme, scienZsts have demonstrated that a geneZc variaZon in the brain makes some people
inherently less anxious, and more able to forget fearful and unpleasant experiences. This lucky geneZc
mutaZon produces higher levels of anandamide the so-called bliss molecule and our own natural
marijuana in our brains. In short, some people are prone to be less anxious simply because they won the
geneZc sweepstakes and randomly got a geneZc mutaZon that has nothing at all to do with strength of
character."

A blogger at ScienZc American

Available @ objected to both the essay and the study, noZng that

"Friedmans [essay] is, in eect, an extremely dumbed down, sensaZonalized press release for a highly
technical arZcle...People are desperate for genuine advances in understanding and treaZng disorders such
as substance abuse and pathological anxiety. Prominent scienZsts such as Richard Friedman and media
such as The New York Times do these people a disservice by oering false hope based on imsy science."
Con@nued

2015)



character."

A blogger at ScienZc American

Available @ objected to both the essay and the study, noZng that

Con@nued

2015)



character."

But a blogger at Scien;c American

Available @
hQp://blogs.scienZcamerican.com/cross-check/2015/03/13/n-y-Zmes-hype-of-feel-good-gene-makes-
me-feel-bad/ objected to both the essay and the study, noZng that

Con@nued

2015)



character."

But a blogger at Scien;c American

Available @
hQp://blogs.scienZcamerican.com/cross-check/2015/03/13/n-y-Zmes-hype-of-feel-good-gene-makes-
me-feel-bad/ objected to both the essay and the study, noZng that

Con@nued

What do you think?

Take a closer look at the 3 documents. Comment on any of the following issues:

Strengths and limitaZons of the study
Sample size
Focus on homologous behavioral and brain phenotypes (markers) in 2 species
Procedures for manipulaZng and measuring anxiety
Were the authors appropriately circumspect and sober in their discussion of the studys
potenZal implicaZons, or did they overreach?
Strengths and weaknesses of the NY Times essay (Over-selling? Dumbed down?)
Strengths and weaknesses of the Scien;c American rebuQal (Too quick to dismiss? Didnt get the
value of combining mechanisZc models in rodents with non-invasive measures of emoZonal
experience and neurophysiology in humans)
The gap between science, the media (NY Times/ScienZc American), and the public. What gets
lost in translaZon?
Con@nued

What do you think?

Take a closer look at any of the 3 documents. Comment on any of the following issues:

Strengths and limitaZons of the study
Sample size
Focus on homologous behavioral and brain phenotypes (markers) in 2 species
Procedures for manipulaZng and measuring anxiety
Were the authors appropriately circumspect and sober in their discussion of the studys
potenZal implicaZons, or did they overreach?
Strengths and weaknesses of the NY Times essay (Over-selling? Dumbed down?)
Strengths and weaknesses of the Scien;c American rebuQal (Too quick to dismiss? Didnt get the
value of combining mechanisZc models in rodents with non-invasive measures of emoZonal
experience and neurophysiology in humans)
The gap between science, the media (NY Times/Scien;c American), and the public. What gets
lost in translaZon?
Con@nued
Time-Permirng
Review Ques;ons
T&P reect
A. Nature
B. Nurture
C. Both
0%
Bo
t
re
0%
Nu
rtu
Na
t
ur
e
0%
Genes (nature) can inuence

A. Environments and
experience
B. Neither. Nature and
nurture are disZnct
and independent
forces
0%
er
. N
at
Ne
ith
En
vir
o
nm
en
t
s a
ur
e a
n
d
nu
nd
e
xp
er
i..
rtu
..
0%
Nature (heritability) is
A. Fixed and
immutable
B. PlasZc and can
change in response
to growing
autonomy or due to
cumulaZve impact
0%
d
ca
n
ic
an
as
t
Pl
Fix
ed

an
d
ch
im
m
ut
an
ge
in
...
ab
le
0%
Heritability is
V
=
G
P
V
al
ot
/
T
GV
0%
nd

/ G
V.
ti.
.
ria
n
of
va
or
tio
e p
ro
p
Th
0%
.
0%
A
a
A. The proporZon of
variaZon in a trait,
such as C/SC, that is
accounted for by
the pedigree (family
tree)
B. PV/GV
C. A and B
EsZmates of heritability
0%
ci.
.
xe
d
0%
Ca
n
be
in
flu
en
ce
d
by
so
Ar
e
fi
A. Are xed
B. Can be inuenced by
social and
environmental
inuences (e.g., living in
a conservaZve religious
community) that
increase or decrease the
amount of variaZon in
the trait (e.g.,
disinhibiZon, partying,
smoking)
Heritability
A. Is the % of variaZon
in a trait, such as E/
PE, that is passed
down from your
parents
B. Reects the
inheritance of
genes, not
phenotypes or traits
0%
e
in
he
rit
a
th
Re
f le
ct
s
Is
t
he
%
o
f v
a
ria
tio
n
in
a
t
nc
e
o
...
...
0%
Heritability describes
ith
in
ty
pi
In
d
ivi
du
als
he
no
o
f p
e %
Th
0%
pu
...
c v
a
s .
..
t i
ha
t t
ra
i
o
f m
y t
e %
Th
0%
ri.
..
0%
p
o
A. The % of my trait that

is inherited (nature)
vs. environmental
(nurture)
B. The % of phenotypic
variaZon across a
group of individuals
that is inuenced by
geneZc factors
C. Individuals within a
populaZon (e.g., Alex)
Highly heritable traits, such as height

A. Are our desZny
B. Can potenZally be
powerfully
inuenced by
intervenZons
(environment)
0%
Ca
n
po
t
en
t
ia
Ar
e
o
lly
b
e
p
ur
d
ow
es
t
er
f..
.
in
y
0%
The End
Things to Consider Adding
SomaZc mosaicism: DNA diers across

neurons
Extra Slides
A few interim take home points

1. Recap: What % of the variance in T&P (phenotype) is due to genes/nature vs.
environment/nurture?
2. What is heritability (h2)? What are the limita;on of this parameter? What are common
misconcep;ons about h2?
3. What has psychiatric gene;cs taught us?
4. What are the long-term prospects for linking heritable traits (e.g., T&P) to dis;nct
neurobiological systems?
5. There is a considerable excitement about so-called neurogene;c approaches that
combine measures of molecular gene;c varia;on (SNPs) with measures of brain func;on
(fMRI). What are the seminal observa;ons? What are some of the key challenges facing
the nascent eld of neurogene;cs?
6. What are G*E interac;ons? How does gene;c inuence depends on the environment?
7. How does the environment get under the skin?
8. What are the 3 kinds of G-E correla;ons? What is the fundamental implica;on of G*E
interac;on and G-E correla;ons for the Nature vs. Nurture dichotomy
What about the brain?
Genome
Intermediate Phenotype
Traits (Evildoing)
Bogdan (Wash U)
The Neurogene;c Strategy

Link gene;c varia;on (polymorphisms) to varia;on in brain structure and func;on (MRI)

Address how genes inuence behavior remember, heritability does not address biological mechanism!
- by correlaZng geneZc variaZon with intermediate biological phenotypes (e.g., amygdala
acZvaZon), we can discover testable mechanisms for geneZc inuence on behavior

Address the molecular mechanisms linking genes to brain to behavior
- its hard to directly measure neurochemistry (e.g., serotonin levels in the amygdala) in humans

- If we measure a geneZc polymorphism with a known funcZon (e.g., serotonin transporter SNP)

- and we are willing to make some assumpZons (dierences in the SNP have predictable eects
on gene expression and ulZmately serotonin levels in the amygdala)

- then we can use geneZc variaZon (polymorphisms), which we can noninvasively measure in
humans, as a proxy for individual dierences in neurochemistry (serotonin in the amygdala),




- can address quesZons such as, Why is the amygdala hyper-reac;ve in behaviorally
inhibited individuals?

Poten;ally address the molecular mechanisms linking genes to brain to behavior

- If we measure a geneZc polymorphism with a known funcZon (e.g., serotonin transporter)

- and we are willing to make some assumpZons (dierences in the poly. have predictable eects

























humans (from peripheral Zssue), as a proxy for individual dierences in neurochemistry
(serotonin in the amygdala),

Seminal Example: Amygdala & 5-HTTLPR

- Threat-related amygdala reacZvity is correlated with variaZon in the
serotonin-transporter linked polymorphic region (5-HTTLPR) on the
SLC6A4 gene
- S allele is bad: Individuals with the less transcripZonally-ecient
short allele (fewer transporter proteins available to clear serotonin
from the synapse) show heightened threat-related amygdala
reacZvity relaZve to individuals with the long allele
- Gene Amygdala: Meta-analyses suggest that the 5-HTTLPR
genotype accounts for 2-5 of the variance in amygdala reacZvity
- Gene Amygdala MDD: Evidence that these geneZcally
conferred dierences in amygdala reacZvity mediate some of the
associaZon between the 5-HTTLPR polymorphism and depression

SLC6A4 gene
reacZvity relaZve to individuals with the long allele
- Gene Amygdala: Meta-analyses suggest that the 5-HTTLPR
genotype accounts for 2-5 of the variance in amygdala reacZvity

SLC6A4 gene
reacZvity relaZve to individuals with the long L allele
- Gene Amygdala: Meta-analyses suggest that 5-HTTLPR accounts
for 2-5% of the variance in amygdala reacZvity

SLC6A4 gene
reacZvity relaZve to individuals with the long L allele
- Gene Amygdala: Meta-analyses suggest that 5-HTTLPR accounts
for 2-5% of the variance in amygdala reacZvity
associaZon between 5-HTTLPR and MDD
The Problem of Assump;ons

These data suggest the following e;ologic chain:
[GENETIC OBSERVATION] 5-HTTLPR

[ASSUMPTION] reduced ecacy of 5HTT (protein)

[ASSUMPTION] too much 5HT in amygdala synapses (chemistry)

[NEURAL OBSERVATION] increased amygdala reacZvity to threat

[EPIDEML OBSERVATION] MDD, especially among individuals exposed
to stress
The Problem of Assump;ons

Kalin (UW)

No rela;on between polymorphism and amygdalar 5HTT expression
when you actually go in and measure the transporter using PET

our ndings are in agreement with the majority of human PET
studiesthat suggest there is not adetectable relaZonship between in
vivo 5-HTT binding and s-allele carrier status our work in the rhesus
monkey, and that of others in humans, calls into quesZon whether this
increased risk is mediated by changes in the expression of the number
of serotonin transporter molecules.
Alex stop here / do epigene;cs next

;me
Brief Aside on How the Environment

Gets Under the Skin

Students:
whats a plausible mechanism?

How might paren@ng or exposure to
other risks inuence behavior
(phenotype)?
How Does E Get Under the Skin?

Epigene;cs provides a biological explana;on for how E (paren;ng, therapy, life events) alters behavior

- The environment (e.g., learning, stress) can alter gene expression (protein synthesis) without altering the
genome (DNA; hence, not heritable)
-
Gene expression is inuenced by transcripZon factors, which bind to sequences of DNA
Binding of transcripZon factors turns genes on or o
EpigeneZc mechanisms involve changes to how readily transcripZon factor can access the DNA
- E.g., methylaZon: addiZon of a methyl group onto a cytosine (1 of the 4 base

pairs that make up DNA) silences the gene because methyl hinders the transcripZon factors

EpigeneZc modicaZons of the genome have long been known to exist e.g., all cells in the body share the
same DNA; accordingly, there must be a mechanism whereby dierent genes are acZve in liver cells vs.
neurons
Work in rodents by Michael Meaneys group demonstrates that maternal behavior can inuence the adult T&P
of ospring and that this is epigeneZc dependent


-


neurons
Work in rodents by Michael Meaneys group demonstrates that maternal behavior (x-fostered) can inuence
the adult T&P of ospring and that this is epigeneZc dependent


-


neurons
Elegant mechanisZc work in rodents by Michael Meaneys group demonstrates that one aspect of the early
environment , maternal behavior (x-fostered), can inuence the T&P of ospring and that this is epigeneZc
dependent
This is exceedingly hard to study in humans because epigeneZc mechanisms vary across the brain and body, so
measuring epigeneZc eects in blood or saliva may not tell you very much about the amygdala
PSYC 612 R08B:

G-E Correla@ons:
How Genes Get Outside the Skin

AJ Shackman
9 December 2013
PSYC 612 R08B:

G-E Correla@ons:
How Genes Get Outside the Skin

Students?
Lemery (ASU)
Jaee (Penn)
G-E Correla;ons Dened (Plomin 77)

Many sources of inuence that we might consider environmental are actually non-random and gene;c

1. Passive G-E correla;on (nature and nurture are confounded)
- among biologically related parents and ospring, the parents provide genotypes AND rearing
environment; thus many parent-child outcome correlaZons may actually reect passive G-E eects
-
E.g., the reason children who are spanked or smacked are more aggressive than children who are not
may be that parents and kids share a geneZc risk for aggressive behavior (common cause)

2. Evoca;ve G-E correla;on
- e.g., a child who is predisposed to having an outgoing, cheerful T&P is more likely to evoke posiZve
aQenZon from others than a child who is predisposed to N/NE
-
E.g., Individuals with a grumpy, abrasive temperament (N/NE) tend to evoke unpleasant responses
from coworkers and others than cheerful, friendly individuals

3. Ac;ve G-E correla;on
- Individuals acZvely select environments
- E.g., individuals predisposed to high E/PE seeking may be more prone to aQend parZes, go to bars, meet
new people, be exposed to or to try substances of abuse


environment; thus many parent-child outcome correlaZons reect passive G-E eects
-

-
E.g., Individuals with a grumpy, abrasive temperament (N/NE) tend to evoke unpleasant responses
from coworkers and others than cheerful, friendly individuals



-

-
E.g., Infant behavioral inhibiZon evokes parental insensiZvity, which then potenZates
maladapZve parentchild interacZons over Zme, exacerbaZng fear of novelty



-

-
E.g., Infant behavioral inhibiZon evokes parental insensiZvity, which then potenZates
maladapZve parentchild interacZons over Zme, exacerbaZng fear of novelty

3. Ac;ve G-E correla;on (Niche Building)
-
E.g., individuals predisposed to high E/PE seeking may be more prone to aQend parZes, go to bars, meet
new people, be exposed to delinquent peers, and try substances of abuse
Evidence for G-E Correla;ons

Mostly from FTA studies demonstra;ng that environmental measures are heritable, including many linked to
psychopathology

e.g., marital quality, social support, parental discipline/warmth, family environment, peer relaZonships,
negaZve life events such as divorce and exposure to trauma

Environments are heritable because genotype inuences behaviors that evoke, select, and modify features of
the environment

- Environments less amenable to behavioral modicaZon are less heritable, e.g., the death of a loved
one, losing ones home in a natural disaster

- Than those that depend on the individuals behavior, e.g., divorce, geng red

Take home: GeneZc risk factors do not necessarily have direct eects on phenotypes (T&P, Dx), but can work
indirectly by modifying sensiZvity to environmental risk factors (acZve G-E) or by inuencing exposure
to risk (passive, evocaZve G-E)


Mostly from FTA studies demonstra;ng that environmental measures are heritable, including many linked to
psychopathology

e.g., marital quality, social support, parental discipline/warmth, family environment, peer relaZonships,
negaZve life events such as divorce and exposure to trauma

Environments are heritable because genotype inuences behaviors that evoke, select, and modify features of
the environment

- Environments less amenable to behavioral modicaZon are less heritable, e.g., the death of a loved
one, losing ones home in a natural disaster

- Than those that depend on the individuals behavior, e.g., divorce, geng red

Take home: GeneZc risk factors do not necessarily have direct eects on phenotypes (T&P, Dx), but can work
indirectly by modifying sensiZvity to environmental risk factors (acZve G-E) or by inuencing exposure
to risk (passive, evocaZve G-E)

G-E Correla;on Take Homes

3 Kinds of G-E Correla;ons: Passive, Evoca;ve, and Ac;ve

Take home: GeneZc risk factors do not necessarily have direct eects
on phenotypes (T&P, Dx), but can work indirectly by modifying
exposure to environmental risks (e.g., stress, substances, delinquent
peers) that reinforce parZcular personality traits or precipitate frank
psychopathology

More fundamentally, these data emphasize that Nature/Genotype and
Nurture/Environment are not mutually exclusive forces, but o|en work
together to increase or decrease the likelihood of important outcomes

Long Term Prospects, The Good

We could get lucky J

In the most opZmisZc scenario, nearly all of the veried risk genes idenZed through GWAS or
sequencing will map to a single coherent inter-connected biological pathway.

This will occur only if the geneZc underpinnings of the disorder reect a high degree of
eZological homogeneity. i.e. a single disease process (equinality)

Intermediate scenarios (neither Ugly nor Good) are possible

Long Term Prospects, The Ugly

There are many many ways to make a neuron hypofunc;on (shrunken
dendrites, too few or dysfuncZonal receptors, downregulated 2nd
messenger systems, or decient transport mechanisms.

There are even more pathways to make a complex
circuit dysfunc;on

This scenario is likely if there are hundreds of disZnct biochemical
changes that individually contribute to Dx (neither necessary nor
sucient) with independent pathways to the phenotype.

Perhaps there are too many ways for the human brain to produce symptoms/signs of
psychiatric disorders (for example, sad mood, auditory hallucinaZons, grandiosity) for a limited
number of biologically coherent pathways to emerge from the 100s or 1000s of genes that
make small contribuZons to risk.

Here, psychiatric disorders (or T&P) arise at such a high level within the mindbrain system
that we have no way to integrate GWAS or sequencing ndings.

If so, then a geneZc mess is a plausible outcome and genes are the wrong level for trying to
understand the biological mechanisms that cause such a trait.
Neurogene;cs Take Homes

1. There is considerable excitement about the neurogeneZcs approach.
2. This reects both clinical interests (Tx) as well as the basic science hope that it can provide clues about the
molecular dierences that inuence the eects seen in fMRI studies (e.g., understand inuence of 5HT
without actually measuring 5HT).
3. But assumpZons may not be warranted; e.g., 5HTTLPR is unrelated to transporter expression in amygdala
4. NeurogeneZcists face all of the problems outlined by Kendler: The Broken Glass, and, The Jet Mechanic. No
guarantee that there are a limited number of funcZonally coherent substrates to be idenZed.
5. The eects of single polymorphisms, such as 5HTTLPR, tend to be weak, necessitaZng large, expensive
samples and begging quesZons about cost/benet.
6. G*E approaches (life stress and 5HTTLPR) have led to much excitement, and may unmask bigger eects and
increased understanding. But at Zmes, it feels like a shing expediZon.
7. Likewise, G*G interacZons (epistasis) and mulZlocus proles that address the aggregate eect of many small-
eect genes may prove helpful, but seem to lead back to the black box of aggregate h2 measures.
8. Combinatorial complexity is daunZng (6M variants!). More sophisZcated modeling and machine learning
approaches will be needed. The primate brain is too stupid to decipher the human brain without help.
9. On the assumpZon that Kendlers Intermediate Scenarios appear most probable, neurogeneZcs appears
useful. Especially when it is integrated with mechanisZc work in nonhuman models (cf. Borsook; Bogdan)
Transla;onal Promise

Sara argues that, in principle, if one could iden;fy with high sensi;vity and specicity at-risk G-E pairs

- At-risk kids paired with risky environments (parental style, peers, adversity, abuse, etc.)

You could target them for precision interven;ons BEFORE the onset of cumula;ve damage

- in eect, she argues for a more nuanced extension of the MoQ PNAS strategy

- instead of idenZfying kids with low C/SC

- idenZfy kids with low C/SC and other environmental risk factors

- this is akin, as I understand it, to what Andreas lab does (ADHD kid + parent with sub-opZmal skill)

- potenZally, one could use biomarkers (gene screens) to idenZfy high-risk parent-kid dyads

More Sophis;cated Approaches:

Gene*Gene & Mul;locus Proles
The phenotype (T&P/Dx) reects the cumulaZve eect of all the genes; traits
are massively polygenic

In principle, it would be helpful to model gene*gene interacZons or develop
more complex addiZve (many main eects) proles (high on this, medium on
that, low on the other and so on)

In pracZce, this is challenging given the combinatorial complexity

Also, prole scores that combine many genes eliminates the possibility of
tesZng specic mechanisZc hypotheses in animal models, back to black box
of aggregate heritability

There is considerable excitement about the development of more
sophisZcated analyZc tools (e.g., machine learning of phenotypically
interesZng gene proles)

More Sophis;cated Approaches:

Gene*Gene & Mul;locus Proles
The phenotype (T&P/Dx) reects the cumulaZve eect of all the genes; traits
are massively polygenic

In principle, it would be helpful to model gene*gene interacZons or develop
more complex addiZve (many main eects) proles (high on this, medium on
that, low on the other and so on)

In pracZce, this is challenging given the combinatorial complexity

Also, prole scores that combine many genes eliminates the possibility of
tesZng specic mechanisZc hypotheses in animal models, back to black box
of aggregate heritability

There is considerable excitement about the development of more
sophisZcated analyZc tools (e.g., machine learning of phenotypically
interesZng gene proles)

Bogdan (Wash U)
The G*E Strategy

- Ryan and Ahmad argues that examining G*E interac;ons is more
realis;c
- insofar as we believe (e.g., the material covered in prior lectures)
that psychopathology and T&P reect the interacZon of
geneZcally endowed diatheses and negaZve life events (e.g.,
stress, adversity, abuse, loss) and learning
- and not the direct consequence or main eect of either G or E
- Ryan argues that the eects are likely to be bigger as well
Some Take Homes

1. There is considerable excitement about the neurogeneZcs approach.
2. This reects both clinical interests (Tx) as well as the basic science hope that it can provide clues about the
molecular dierences that inuence the eects seen in fMRI studies (e.g., understand inuence of 5HT
without actually measuring 5HT).
3. But assumpZons may not be warranted; e.g., 5HTTLPR is unrelated to transporter expression in amygdala
4. NeurogeneZcists face all of the problems outlined by Kendler: The Broken Glass, and, The Jet Mechanic. No
guarantee that there are a limited number of funcZonally coherent substrates to be idenZed.
5. The eects of single polymorphisms, such as 5HTTLPR, tend to be weak, necessitaZng large, expensive
samples and begging quesZons about cost/benet.
6. G*E approaches (life stress and 5HTTLPR) have led to much excitement, and may unmask bigger eects and
increased understanding. But at Zmes, it feels like a shing expediZon.
7. Likewise, G*G interacZons (epistasis) and mulZlocus proles that address the aggregate eect of many small-
eect genes may prove helpful, but seem to lead back to the black box of aggregate h2 measures.
8. Combinatorial complexity is daunZng (6M variants!). More sophisZcated modeling and machine learning
approaches will be needed. The primate brain is too stupid to decipher the human brain without help.
9. On the assumpZon that Kendlers Intermediate Scenarios appear most probable, neurogeneZcs appears
useful. Especially when it is integrated with mechanisZc work in nonhuman models (cf. Borsook; Bogdan)
Strategy for Linking FTA to Molecules

Dick notes that it is useful to rst search for G*E interac;ons in FTA studies

There are a huge number of environmental factors that could potenZally be assessed

Useful to build o the already well-developed developmental literature

In cases where there is an interacZon between Genes (in aggregate) and Environment (e.g., peer delinquency
exposure), that is, a hit in the FTA literature

It makes sense to drill down into specic genes, either candidate variants or GWAS
R08A/B Key Learning Objec;ves

1. Recap: What % of the variance in T&P (phenotype) is due to genes/nature vs.
environment/nurture?
2. What is heritability (h2)? What are the limita;on of this parameter? What are common
misconcep;ons about h2?
3. What has psychiatric gene;cs taught us?
4. What are the long-term prospects for linking heritable traits (e.g., T&P) to dis;nct
neurobiological systems?
5. There is a considerable excitement about so-called neurogene;c approaches that
combine measures of molecular gene;c varia;on (SNPs) with measures of brain func;on
(fMRI). What are the seminal observa;ons? What are some of the key challenges facing
the nascent eld of neurogene;cs?
6. What are G*E interac;ons? How does gene;c inuence depends on the environment?
7. How does the environment get under the skin?
8. What are the 3 kinds of G-E correla;ons? What is the fundamental implica;on of G*E
interac;on and G-E correla;ons for the Nature vs. Nurture dichotomy
Goldsmith Slides
2. Two major types of genomic variants

1. Sequence variants
-SNPs
-single insertions & deletions
2. Structural variants (major categories)

-tandem repeats (also called microsatellites)
-triplet repeats (also called minisatellites)
-CNVs (copy number variants), to be
discussed soon
The genome
3.1-3.2 billion
bases
~21-23,000 genes

The most common type of genetic

variantthe SNP
SNP = single nucleo;de polymorphism
usually diallelic (2/3rd are C/T)
SNPs account for 90% of human genetic
variation
In the human genome there are an estimated
10,000,000 - 11,000,000 common SNPs (with
> 1% frequency) (dbSNP)
Occur once in 300 bases
~7,000,000 SNPs have been identified and
provisionally mapped

Microsatellite Markers: e.g., Tandem Repeats

ATTGTTATCCGCTCACAATTCCACACAAT
CACACACACACACACACACACACAGAGTGAGCTAACTCACATTAATTGCGTTGC
6 repeat units
ATTGTTATCCGCTCACAATTCCACACAAT
CACACACACACACACACACACACACACACACACACAGAGTGAGCTAACTCACATTAATTGCGTTGC
9 repeat units
frequently found within the genome
highly polymorphic
Minisatellite Markers: 10-100 bp repeats (e.g., in Fragile X)

-variaZon in the number of copies is likely due to neighboring
cis-acZng meioZc double-strand break hotspots
Copy Number Variants (CNVs)

A structural variant in the DNA rather than a sequence
variant (such as a SNP)
Regions of genomic DNA (>1000 bases in length, up to 5
megabases) that can be duplicated or deleted
The Human Genome Project showed copy number
variaZon to be common. A comparison of 2 individuals
showed 297 possible CNV dierences across the genome.
CNVs can occur within one gene or involve >1 gene. Thus,
CNVs involving dosage-sensiZve genes can have funcZonal
signicance.
CNVs may be especially important in speciaZon (human vs.
chimp dierences in CNVs are prominent)
Large and rare CNVs seem prominent in neurocogniZve
disorders
3. Three stages of human behaviorgenetic investigation

1. Quantitative genetic approaches to estimate
overall genetic variance
-Family (pedigree), twin, & adoption designs
2. Gene discovery approaches (this week)
3. Gene environment interplay, and other
functional investigations with identified genes
(last unit of course)
-Overlap and integration of approaches; e.g., twin
designs used in all 3 approaches, esp. #1 & #3
4. The two major classifications of disorders

that determine which methods of gene
identification should be pursued
21 rare alleles
3 common alleles

Common Disease-Common
Variant (CDCV)
Models of Common Inherited Disease
Risk alleles are common, > 5%

SelecZon is weak
Risk mutaZons are old
Limited # of alleles
oligogenic
Risk alleles have weak to
moderate eects on
phenotypes
Risk alleles are shared across
populaZons

Common Disease-Rare
Variant (CDRV)
Risk alleles are rare, < 1%

SelecZon is strong
Risk mutaZons are recent
Large # of alleles geneZc
heterogeneity
Range of strengths of eects
Risk alleles are populaZon-
specic

Out of Africa
5. The three major methods of gene

discovery/identification
A. Linkage
B. Allelic Association
-Direct
-Indirect
Next, linkage
Gene Iden;ca;on
Where to look
Targeted
Linkage or CytogeneZc PosiZonal candidate
Pathophysiology Candidate gene
Animal models
Genome-wide
How to look
A. Linkage analysis
B. AssociaZon Studies
Gene;c linkage two loci that reside physically

near each other on the same chromosome.
Linkage can result in a within-family associaZon
between a geneZc marker and disease status
Gene;c marker polymorphic DNA sequence
(typically not funcZonal) of known chromosomal
locaZon
Recombina;on -- rate at which disease status is
assoc. with dierent geneZc marker status in
ospring than parents (range is 0% to 50%)
A. Linkage our experience with

linkage so far in this course
An intui;ve example of linkage

analysis for a polygenic trait
- The aected sib pair method,
(analyzing allele sharing)
Sample, disorder, set of markers
Linkage Analysis: formal

deni;on
StaZsZcal procedure for analyzing within-
family associaZons between geneZc
markers and disease phenotypes that takes
into account the complicaZons of reduced
penetrance and crossing-over.
Test staZsZc = lod score
Lod Score (the measure of linkage)

lod = log10 (odds[linkage:no linkage])
odds = probability linked at ( <.50)
probability unlinked ( =.50)
(theta is the recombinaZon frequency between the marker
and unknown disease gene; theta can be tested at values
between 0 and .50)
lod is considered signicant if > 3.0 (region

posiZve) or < -2.0 (region excluded)
Lod Scores
Lod Score
Odds for Linkage
-3
1:1000
-2
1:100
-1
1:10
10:1
100:1
1000:1
An early linkage study:

Sherrington et al. (1988)
Targeted 5q11-5q13 in 7 pedigrees (104
individuals with 44 aected)
Assumed schizophrenia was autosomal
dominant with 86% penetrance.
Lod score = 6.5 at = .08 (odds beQer
than 3,000,000:1).
Genome-Wide Linkage Analysis

Targeted Linkage Approach
Depends on fortuitous observaZons
Genome-Wide linkage Approach

Need a marker AT LEAST every ~ 10cM
300-400 markers
Summary on Linkage Analysis

Advantages:
Feasible to implement genome-wide (systemaZc &

comprehensive); dont need to have hypotheses
about locaZon or mechanism
As compared to alternaZves, relaZvely low false
posiZve rate
Disadvantages
Requires family-study design
Good for genes of large eect; low power to detect
genes accounZng for 5% or less of variance
Limited resoluZon: Zght linkage can be millions of
bases away
Does not nd the gene, rather idenZes a region
B. Allelic Association
AssociaZon between allele status and
phenotype in unrelated individuals.
Popula;on-level associa;on
Stomach Cancer
OR = 60*60 = 2.25
40*40
Not O
(A,B,AB)
O
60
40
40
60
Challenges of Gene Identification

of the 166 putaZve associaZons
which have been studied three or
more Zmes, only 6 have been
consistently replicated.

Hirschhorn et al. (2002). A comprehensive
review of geneZc associaZon studies.
Gene;cs in Medicine 4:45-61.

Genetic Association Strategies

Non-targeted:
genome-wide
Targeted:
Chromosomal anomaly (e.g., VCFS)
Biologically based hypotheses (e.g., DRD4 and
Novelty-seeking)
Animal models
Posi;ve linkage results
Expression studies
The Age of Discovery: A Gene For ...

Date
New York Times Headline
2-26-1986
Defective gene tied to form of manic-depressive illness
11-10-1988
Schizophrenia study finds strong signs of hereditary cause
4-18-1990
Scientists see linkage between alcoholism and a single

gene
7-16-1993
Report suggests that homosexuality is linked to genes
10-22-1993
Gene tie to male violence is studied
1-02-1996
Variant gene tied to love of new thrills
And the Age of Loss ...

Date
New York Times Headline
2-26-1986
1-13-1993
Defective gene tied to form of manic-depressive illness

Scientists now say they cant find gene for manic-depressive illness
11-10-1988
11-07-1989
Schizophrenia study finds strong signs of hereditary cause

Scientists now doubt they found faulty gene linked to mental illness
4-18-1990
Scientists see linkage between alcoholism and a single gene
7-16-1993
4-23-1999
Report suggests that homosexuality is linked to genes

Study questions gene influence on homosexuality
10-22-1993
2-14-1992
Gene tie to male violence is studied

Genes dont doom anyone to a life of crime
1-02-1996
11-01-1996
Variant gene tied to love of new thrills

Maybe its not a gene behind a persons thrill-seeking ways
Allelic Association
Advantages
Easy design to implement
Compares cases versus controls
In principle, very high staZsZcal power
In principle, can idenZfy causal agent
Disadvantages
Need to know funcZonal polymorphisms in a
candidate gene (for direct associaZon)
Concern about false-posiZves due to mis-matching
cases and controls (straZcaZon bias next slide)
Ethnic Group #1: OR = 1.0

- Hyper
+ Hyper
24
(.40)
16
(.40)
40
(.40)
Not
O
36
24
60
60
40
100
- Hyper
+ Hyper
24
(.40)
16
(.40)
40
(.40)
9
(.10)
1
(.10)
10
(.10)
Not
O
36
24
60
Not
O
81
90
60
40
100
90
10
100
- Hyper + Hyper

- Hyper
+ Hyper
24
(.40)
16
(.40)
40
(.40)
Not
O
36
24
60
60
40
100
Combined:
OR = 1.83

- Hyper
+ Hyper
9
(.10)
1
(.10)
10
(.10)
Not
O
81
90
90
10
100
- Hyper
+ Hyper
33
(.22)
17
(.34)
50
(.25)
Not
O
117
33
150
150
50
200
Why the False-Positive Findings?

Stra;ca;on bias (ethnic mismatching)
Winners Curse
IniZal signicant ndings overesZmate the
magnitude of the eect
Cumula;ve Odds Ra;o as a Func;on of Publica;on Year
Original
OR=8.7
Smith et al. (2008) American Journal of Epidemiology, 167(2): 125-138.
Cumula;ve Odds Ra;o as a Func;on of

Publica;on Year
Pooled 1st &
2nd OR= 3.9
Original
OR=8.7
Cumula;ve Odds Ra;o as a Func;on of

Publica;on Year
Original
OR=8.7
Winners Curse
Cumula;ve Odds Ra;o as a Func;on of Publica;on Year
Original
OR=8.7
Final OR=1.4
Why the False-Posi;ve Findings?

Ethnic mismatching
Winners Curse
GeneZc eects smaller than previously thought
Gene-environment Interac;on
G eect may only exist in certain environments
(reminder: heritability esZmates are specic to the
environment in which they are esZmated)

Why the False-Posi;ve Findings?

Ethnic mismatching
Winners Curse
GeneZc eects smaller than previously thought
Gene-environment Interac;on
Eect may only exist in certain environments

APOE
Polymorphism

Allele-specic Odds Ra;os (ORs) for Polymorphisms Signicantly

Associated in Meta-Analyses of Alzheimers Disease
Recap: Finding genetic effects has

been difficult
Expected eect sizes overes;mated
Winners curse non-replicaZon
Sample sizes were too small
Meta-analyses likely give more accurate esZmates
Assump;on that gene;c variants are common

may not always be accurate
Returning to the big picture of

gene identification
Non-targeted, genome-wide approaches
Possible with linkage
UnZl recently, not possible with associaZon
Early-stage strategy: generates candidates
Targeted approaches (e.g., candidate gene)

Require specic hypotheses (someZmes driven
by feasibility more than science)
Can be a late-stage strategy that conrms
candidates from genome-wide approaches
Returning to the 3 approaches

Linkage analysis: within family associaZon
between marker and phenotype
GeneZc eects may not be strong enough
Allelic (Direct) Associa;on: populaZon associaZon
between presumed causal variant and phenotype
May not know enough for specic hypotheses
Thus, we o|en must rely on
Linkage Disequilibrium, or Indirect Associa;on:
PopulaZon associaZon between geneZc marker
and phenotype
Indirect and Direct Allelic Associa;on

Direct Associa;on
D
Assess relaZonship of D locus
to phenotype directly
expect D to be a funcZonal
polymorphism in a candidate gene
Indirect Associa;on
M1 M2 D
M3
Assess relaZonship of D locus

indirectly by determining whether
markers (Mi) are associated with
disease Mi dont need to be
funcZonal
Haplotypes
Haplotype is a generalizaZon of the concept
of a single genotype to mulZple linked loci
refers to a series of markers (usually SNPs)
Haplotypes
Haplotype is a generalization of the
concept of genotype to multiple loci
Recombination is not random, but rather
tends to occur at hot spots
This gives rise to blocks of DNA
(haplotypes), where there is very little
recombination within blocks but extensive
recombination between blocks
STOPPED HERE
Haplotypes
A
3 Genotypes:
AT at locus 1
CG at locus 2
GC at locus 3
Haplotypes (alleles
inherited together on
a segment of the
same chromosome)
ACG & TGC
Haplotype Blocks
Cardon & Abecasis (2003). Using haplotype blocks to map the human genome.
Trends in Gene@cs, 19: 135-140.
Although 9 markers in block 4,

only 4 possible haplotypes, which
can be determined by only 3 markers
Cardon & Abecasis (2003). Using haplotype blocks to map the human genome.
Trends in Gene@cs, 19: 135-140.
Linkage Disequilibrium Mapping

Because of linkage disequilibrium,
inheriZng a marker allele might be a
(imperfect) indicaZon of having inherited a
chromosomal segment with a certain
mutaZon. This will depend on
The number of genera;ons since introducZon
of the original mutaZonal event
How Zghtly linked the marker allele and the
certain mutaZon are
Out of Africa
Empirical Evidence
CDCV model & LD
If variant is common and not selecZvely

advantageous, it must be ancient.
That is, the mutaZon probably existed
when all humans were in Africa (maybe
2500-4000 generaZons ago).
LD may only extend up to 3 kb would
need about 500,000 markers at this spacing
to cover the genome.
Illumina 1M DNA Analysis BeadChip
Lessons From Height

Twin studies high heritability (~ 80-90%)
Genes (mendelian) causing extreme stature
are known
Genes (polygenic) causing normal height
variaZon were not known
Lessons From Height

Despite high heritability esZmates the results have been
disappoin;ng and inconsistent, with reports of
quanZtaZve trait loci (QTLs) scaQered across the genome
and rarely replicated.

Perola, M. et al. (2007). PLOS Gene;cs, 3(6): 1019.
Note the dates of papers/quotes
Lessons From Height

In the past 18 months, the rst robust common variant
associaZons were idenZed and there are now 44 loci
known to inuence normal variaZon in height.

Weedon, M.W. & Frayling, T.M. (2008). Trends in
Gene;cs, 24: 595-603.
Replicated GWAS for Height

(Total N > 160,000)
Study
Original
N
Replica;on
N
Signicant
Findings
Weedon (2007)
4,921
29,098
1 variant
Sanna (2008)
6,669
28,801
1 variant
Weedon (2008)
13,655
16,482
20 variants
Gudbjartsson (2008)
29,820
8,541
27 variants
Lecre (2008)
15,821
> 10,000
12 variants
Average Eect of Replicated Gene;c

Variant = ~0.3-0.4 cm
% Height Variance Accounted For
Goldstein, D.B. (2009). Common gene;c varia;on and human traits. NEJM, 360:
1696-1698
the latest on GWAS of Height

Yang et al. (2010) take a classical quantitative genetic
approach to this problem by using genome-wide genotypes to
infer the genetic relatedness of individuals and then estimate
the relationship of this quantity to phenotypic similarity, thus
arriving at an estimate of heritability explained.
Their model complex trait is height, which has heritability of
~80%, only approximately 5% of which has been explained
by 50 common polymorphisms identified by GWAS.
Starting with a Brisbane, Australia cohort of 3,925 unrelated
individuals (who have less genetic relatedness than typical
second cousins), they in effect linearly regress all of the SNPs
from their GWAS onto an appropriately adjusted measure of
height.
The raw estimate that this produces is that 45% of the total
variation for height (and hence 56% of the heritability) in the
Brisbane cohort can be explained by common SNPs (regardless
of their significance).
By accounting for imperfect linkage disequilibrium between
tagging and causal variants, Yang et al. up their estimate of
common variant effects from 56% to at least 67% of the genetic
contribution to height.
A further correction assumes that causal variants are likely to be
deleterious and hence at lower frequency than tagging SNPs,
and in this case leads to the conclusion that most of the
heritability for height can be captured by common variants.
This presents an elegant argument that most of the

heritability is hidden rather than missing and, hence, that
there is no pressing need to invoke more complex genetic
mechanisms to explain height. Whether or not this is also
true of susceptibility to common diseases remains to be
determined.
Furthermore, there is the caveat that this interpretation of the
result is a parsimonious one and that a variety of genetic
models could also produce similar regressions of phenotypic
on genetic relatedness.
Most commentary from Gibson, 2010, Nature Genetics
MAF & Odds Ra;os of GWAS

Associa;ons
Median =
1.33
Hindor, L. A. et al. (2009). Poten;al e;ologic and func;onal implica;ons of

genome-wide associa;on loci for human diseases and traits. Proceedings of the
Na@onal Academy of Sciences of the United States of America, 106(23), 9362-9367.
GWAS appears successful as newer,

larger studies are reported
18 loci in sample of 249,796
account for 1.5% of variance
in BMI
account for ~10% of variance
in height

account for 3.6-6.1% of variance
in age at menarche
GWAS of Behavioral Traits
As of Jan., 4, 2010 at: hcp://www.genome.gov/GWAStudies
Design:
~3300 Sz and 3600 Controls
1M SNP markers
Results:
Most highly associated SNP
had p = 3.4 x 10-7
Second most highly associated
SNP in MHC region
http://www.genome.gov/GWAStudies/index.cfm?pageid=26525384#searchForm
Maher, B. (2008). Personal genomes: The case of the missing

heritability. Nature, 456(7218), 18-21.
The 5 most probable reasons that GWAS have

failed to account for the missing heritability
Studies are not powerful enough, re sample
sizes and number of markers; found genes that
are not signicant are important
G x G interacZons (epistasis) and G x E
interacZons prevalent, but studies are not
powered to nd these interacZons
GeneZc heterogeneity is common
phenotype deniZon is poor (esp for behavioral
phenotypes)
Many of the important variants are not
common (CVCD model is wrong)
If heterogeneity is common,
Then, rare variants (which would be
populaZon specic) might explain part of
the heritability not explained by common
variants.

Common Disease-Common
Variant (CDCV)
Models of Common Inherited Disease
Risk alleles are common > 5%

SelecZon is weak
Risk mutaZons are old
Limited # - oligogenic
Weak to moderate eects
Shared across populaZons
Common Disease- Rare

Variant (CDRV)
Risk alleles are rare < 1%

SelecZon is strong
Risk mutaZons are recent
Large # - heterogeneity
Range of eects
Popula;on specic
Recurrent CNVs
McCarthy, S. E., Makarov, V., Kirov, G., Addington, A. M., McClellan, J., Yoon, S., et al. (2009).
Microduplica;ons of 16p11.2 are associated with schizophrenia. Nature Gene@cs, 41(11),
1223-1227.
Conclusions: Gene Iden;ca;on

Common Disease, Common Variant:
Can now do genome-wide studies

Eect of any variant will be very small
Require large samples, consorZa & meta-analysis
Clinical implicaZons uncertain
Common Disease, Rare Variant:

Cannot do genome-wide associaZon studies
Rare variants could have large eects
Require large, evoluZonarily homogenous populaZons
(e.g., Finland)
Could have signicant clinical implicaZons
Type of
Association
Interpretation
Genome-wide
Resolution
Preferred Sample
Sensitivity
Specificity
Linkage
Allelic
Association
(Direct)
w/i Family
between
marker and
phenotype
Pop assoc btw

functl poly and
phenotype
Linkage
Disequilibrium
(Indirect)
Pop assoc btw
marker and
phenotype
Allelic
Association
(Direct)
Linkage
Disequilibrium
(Indirect)
Within family
between
marker and
phenotype
Pop assoc btw functl

poly and phenotype
Pop assoc btw

marker and
phenotype
Region
implicated
Gene or nearby
gene
implicated
Region
implicated
Linkage
Type of Association
Interpretation
Genome-wide
Resolution
Preferred Sample
Sensitivity
Specificity
Linkage
Disequilibrium
(Indirect)
Linkage
Allelic Association
(Direct)
Type of Association
w/i Family between

marker and
phenotype

poly and phenotype
Pop assoc btw

marker and
phenotype
Interpretation
Region implicated
Gene or nearby gene

implicated
Region implicated
No, maybe in
the future
(~60,000)
Now feasible
(~500,000,
HapMap)
Genome-wide
approach?
Resolution
Preferred Sample
Sensitivity
Specificity
Yes, 300-400
markers
Linkage
Disequilibrium
(Indirect)
Linkage
Allelic Association
(Direct)
Type of Association
w/i Family between

marker and
phenotype

poly and phenotype
Pop assoc btw

marker and
phenotype
Interpretation
Region implicated
Gene or nearby gene

implicated
Region implicated
Genome-wide
Yes, 300-400 markers
Not now but maybe

in the future
(~60,000)
Now feasible
(~500,000, HapMap)
Resolution
Preferred Sample
Sensitivity
Specificity
Typically w/i
8-10
centimorgans
recombination
Not relevant
Typically
<< 1
centimorgan
Linkage
Disequilibrium
(Indirect)
Linkage
Allelic Association
(Direct)
Type of Association
w/i Family between

marker and
phenotype

poly and phenotype
Pop assoc btw

marker and
phenotype
Interpretation
Region implicated
Gene or nearby gene

implicated
Region implicated
Genome-wide
Not now but maybe

in the future
(~60,000)
Now feasible
(~500,000, HapMap)
Resolution
Typically w/i 8-10

centimorgans
recombination
Not relevant
Typically (much) less

than 1 centimorgan
Unrelated
individuals
Genetic isolates
if rare
Preferred
Sample
Sensitivity
Specificity
Large pedigrees
Sib pairs
Linkage
Disequilibrium
(Indirect)
Linkage
Allelic Association
(Direct)
Type of Association
w/i Family between

marker and
phenotype

poly and phenotype
Pop assoc btw

marker and
phenotype
Interpretation
Region implicated
Gene or nearby gene

implicated
Region implicated
Genome-wide
Not now but maybe

in the future
(~60,000)
Now feasible
(~500,000, HapMap)
Resolution
Typically w/i 8-10

centimorgans
recombination
Not relevant

than 1 centimorgan
Preferred Sample
Large pedigrees
Sib pairs
Unrelated
individuals
Genetic isolates
if rare
Sensitivity
Relatively low
Relatively High
Relatively High
Specificity
Linkage
Disequilibrium
(Indirect)
Linkage
Allelic Association
(Direct)
Type of Association
w/i Family between

marker and
phenotype

poly and phenotype
Pop assoc btw

marker and
phenotype
Interpretation
Region implicated
Gene or nearby gene

implicated
Region implicated
Genome-wide
Not now but maybe

in the future
(~60,000)
Now feasible
(~500,000, HapMap)
Resolution
Typically w/i 8-10

centimorgans
recombination
Not relevant

than 1 centimorgan
Preferred Sample
Large pedigrees
Sib pairs
Unrelated
individuals
Genetic isolates
if rare
Sensitivity
Relatively low
Relatively High
Relatively High
Specificity
Relatively High
Relatively Low
Relatively High
(?)
McCarthy, M. I., et al. (2008). Genome-wide associa;on studies for complex traits: consensus, uncertainty
and challenges. Nature Reviews Gene@cs, 9(5), 356-369.
END OF PRESENTATION
My Friend, Dr. Dan Grupe (~6 5)

Dylan & I are standing on the NEXT level
Yes, from the floor it looked as though Dans head would touch the ceiling
Anx & Dep Assoc of America Annual Meeting 2014
The Case of Height

Thought-experiment:
A sample of 130 000 people would contain 910 cases (tall folks like
Dan)

A casecontrol study of 910 cases and 910 controls for being tall would
have no power to detect any of the reported ndings for height

You would expect to detect none of the variants contribuZng to height
because of insucient staZsZcal power

Small single-nucleoZde eects of the kind that can be detected on SNP
chips mandate large samples, but provide novel insights into the
underlying biological pathways that contribute to trait like individual
dierences in T&P (N, E, C), intermediate phenotypes (amygdala,
ventral striatum), or human diseases, including psychiatric disorders
associated with T&P
Could we run fewer subjects and save

money if we used
a case-control design?
The Case of Height

Thought-experiment:
Taking individuals about 2.5 SDs above the mean (~65 for men), then
~0.7% of the populaZon can be diagnosed with tallness (99.3% < 65)

The Case of Height

Thought-experiment:
A sample of 130,000 people would contain 910 cases (tall folks)

have no power to detect any of the reported ndings for height

StaZsZcally, you would expect to detect none of the variants owing to
insucient power

chips mandate large samples, but provide insights into the underlying
biological pathways that contribute to trait like individual dierences in
T&P (N, E, C), intermediate phenotypes (amygdala, ventral striatum), or
human diseases, including psychiatric disorders associated with T&P
The Case of Height

Thought-experiment:
A sample of 130,000 people would contain 910 cases (tall folks)

Dan Grupe
have no power to detect any LilAlex
of the rBig
eported
ndings for height

StaZsZcally, you would expect to detect none of the variants owing to
insucient power

chips mandate large samples, but provide insights into the underlying
biological pathways that contribute to trait like individual dierences in
T&P (N, E, C), intermediate phenotypes (amygdala, ventral striatum), or
human diseases, including psychiatric disorders associated with T&P
The Case of Height

Thought-experiment:
A sample of 130,000 people would contain 910 cases (tall folks like
Dan)

A casecontrol study of 910 cases and 910 matched controls would
have no power to detect any of the reported geneZc variants for height

Conclusion: You cannot cut-corners using case-control designs
Conceptual Roadmap
Individual dierences in T&P reect the brain

e.g., Individuals with higher levels of N/NE
show enhanced ac;va;on and slower recovery
in the amygdala

Where do dierences in brain ac@va@on come
from?

We cant assay @ssue from the amygdala in
people, but can we use the genome (DNA) to
iden@fy candidate mechanisms that could be
mechanis@cally tested in animals?
Students

What are your fundamental ques;ons about the
Nature and Nurture of individual dierences in
Temperament and Personality?

What do you want to know?

Charles Darwin

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We

wish to suggest a structure for the salt of

we should remain unabashed about the ul;mate

Plan for Today

Take-home cri;cal thinking ques;ons

Quick Gene;cs Primer

What about genes?

- Gene: a region of DNA/RNA sequence, corresponding to a unit of

- Gene: a region of DNA/RNA sequence, corresponding to a unit of

- Gene: a region of DNA/RNA sequence, corresponding to a unit of

- Gene: a region of DNA/RNA sequence, corresponding to a unit of

What makes us dierent?

Alleles = Individual Dierences

Alleles = Individual Dierences

Alleles = Individual Dierences

Alleles = Individual Dierences

SNPs = Individual Dierences

SNPs = Individual Dierences

How might we try to discover the

Lets start with a

Allele #1 is associated with a 9x

hQp://vassarstats.net/odds2x2.html (0.90 allele #1 = paZents; 0.10 allele #2 = paZent; 0.90/0.10 = 9 )

Now we need to scale this up

How the heck are we going to

Details Are Not Important

Details Are Not Important

Fragment of one strand

Details Are Not Important

Fragment of one strand

Binds with complement

Details Are Not Important

Fragment of one strand

Binds with complement

Array of probes for

Fragment of one strand

Binds with complement

Fragment of one strand

Binds with complement

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

Enhance prognosis: You have 3 months to live

Monitor treatment in terms of changes in the underlying neural systems

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

Enhance prognosis: You have 3 months to live

Monitor treatment in terms of changes in the underlying neural systems

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

What is the biological basis of childhood shyness?

IdenZfy at-risk individuals early (e.g., carriers of a parZcular polymorphism)

What is the biological basis of childhood shyness?

Some Success with GWAS

Some Success with GWAS

Some Success with GWAS

Some Success with GWAS

But theres a poten@al problem