Nincs diacm

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Genetic Association Study of the Dopamine D4
receptor gene polimorphism and REACTION TIME
PERFORMANCE in normal subjects
Anna Szekely, Ph.D. Dr. Zsolt Ronai, Dr. Zsofia
Nemoda Dr. Maria Sasvari-Szekely
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The story of the DRD4-VNTR 7
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The Human Genomee Project
GTCCGGTCCC GGGACCCCCT GCCCAGGGTC AGAGGGGCGC
CTACCTAGCT CACGGTCTTG GGCCGGAGGG AATGGAGGAG
GGAGCGGGGT CGACCGCTCA GCTGTCCGCC
CAGTTTCGGA GGCGGCCACG CGAGGATCAA CTGTGCAACG
GGTGGGGCCG CGGCTGACCG TGGTGGTCGC GGGGGCTGAG
GGCCAGAGGC TGCGGGGGGG GGGCGGCGGG ATGAGCTAGG
CGTCGGCGGT TGAGTCGGGC GCGGAGTCGG GGGCAGGGGG
AGCGGGCGTG GAGGGCGCGC ACGAGGTCGA GGCGAGTCCG
CGGGGGAGGC GGGCAGAGCC TGAGCTCAGG TCTTTCTGCG
TCTGGCGGAA CGGGCCTGGG AGGGAGGTTT TGCCAGATAC
CAGGTGGACT AGGGTGAGCG CCCGAGGGCC GGGACGCACG
CACGGGCCGG GTAGGATGGC GCTGGCGTCG ATGCCCGCGC
GCTTCAGGGC CTGGTCTGGC CGCCCCTCCA TCCTTGTCGG
TTTCTCGGGT CGCGGACCCC GCGCGGCGCC GGGCGATGCT
GGCCTGCCCG TGGCCACCAC CTCGCTTCAT TCCCGTCTCT
TTGGGCCGCC GCATTCGTCC ACGTGCCCGT CTCTCCCTGC
GCAAAATTCC AAGATGAGCA AATACTGGGC TCACGGTGGA
GCGCCGCGGG GGCCCCCCTG AGCCGGGGCG GGTCGGGGGC
GGGACCAGGG TCCGGCCGGG GCGTGCCCGA GGGGAGGGAC
TCCCCGGCTT GCGACCCGGC GTTGTCCGCG
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A Humán Genome Projekt
GTCCGGTCCC GGGACCCCCT GCCCAGGGTC AGAGGGGCGC
CTACCTAGCT CACGGTCTTG GGCCGGAGGG AATGGAGGAG
GGAGCGGGGT CGACCGCTCA GCTGTCCGCC
CAGTTTCGGA GGCGGCCACG CGAGGATCAA CTGTGCAACG
GGTGGGGCCG CGGCTGACCG TGGTGGTCGC GGGGGCTGAG
GGCCAGAGGC TGCGGGGGGG GGGCGGCGGG ATGAGCTAGG
CGTCGGCGGT TGAGTCGGGC GCGGAGTCGG GGGCAGGGGG
AGCGGGCGTG GAGGGCGCGC ACGAGGTCGA GGCGAGTCCG
CGGGGGAGGC GGGCAGAGCC TGAGCTCAGG TCTTTCTGCG
TCTGGCGGAA CGGGCCTGGG AGGGAGGTTT TGCCAGATAC
CAGGTGGACT AGGGTGAGCG CCCGAGGGCC GGGACGCACG
CACGGGCCGG GTAGGATGGC GCTGGCGTCG ATGCCCGCGC
GCTTCAGGGC CTGGTCTGGC CGCCCCTCCA TCCTTGTCGG
TTTCTCGGGT CGCGGACCCC GCGCGGCGCC GGGCGATGCT
GGCCTGCCCG TGGCCACCAC CTCGCTTCAT TCCCGTCTCT
TTGGGCCGCC GCATTCGTCC ACGTGCCCGT CTCTCCCTGC
GCAAAATTCC AAGATGAGCA AATACTGGGC TCACGGTGGA
GCGCCGCGGG GGCCCCCCTG AGCCGGGGCG GGTCGGGGGC
GGGACCAGGG TCCGGCCGGG GCGTGCCCGA GGGGAGGGAC
TCCCCGGCTT GCGACCCGGC GTTGTCCGCG
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Similarity in terms of gene sequence
Unrelated humans share - 99.9 (the difference
is about 3 x 106 bp) Mutations Polimorphisms
GAGGGTGCGC
GAGGGAGCGC
GAGGGAGCGC
GAGGGTGCGC
Human apes share 95
GAGGGTGCGC
GAGGGAGCGC
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when the misprint is fatal
Mutations rare allele variations - usually
monogenic disorders (in less than 1 of the
human population)
GTCCGGTCCC GGGACCCCCT GCCCAGGGTC AGAGGGGCGC
CTACCTAGCT CACGGTCTTG GGCCGGAGGG AATGGAGGAG
GGAGCGGGGT CGACCGCTCA GCTGTCCGCC
CAGTTTCGGA GGCGGCCACG CGAGGATCAA CTGTGCAACG
GGTGGGGCCG CGGCTGACCG TGGTGGTCGC GGGGGCTGAG
GGCCAGAGGC TGCGGGGGGG GGGCGGCGGG ATGAGCTAGG
CGTCGGCGGT TGAGTCGGGC GCGGAGTCGG GGGCAGGGGG
AGCGGGCGTG GAGGGCGCGC ACGAGGTCGA GGCGAGTCCG
CGGGGGAGGC GGGCAGAGCC TGAGCTCAGG TCTTTCTGCG
TCTGGCGGAA CGGGCCTGGG AGGGAGGTTT TGCCAGATAC
CAGGTGGACT AGGGTGAGCG CCCGAGGGCC GGGACGCACG
CACGGGCCGG GTAGGATGGC GCTGGCGTCG ATGCCCGCGC
GCTTCAGGGC CTGGTCTGGC CGCCCCTCCA TCCTTGTCGG
TTTCTCGGGT CGCGGACCCC GCGCGGCGCC GGGCGATGCT
GGCCTGCCCG TGGCCACCAC CTCGCTTCAT TCCCGTCTCT
TTGGGCCGCC GCATTCGTCC ACGTGCCCGT CTCTCCCTGC
GCAAAATTCC AAGATGAGCA AATACTGGGC TCACGGTGGA
GCGCCGCGGG GGCCCCCCTG AGCCGGGGCG GGTCGGGGGC
GGGACCAGGG TCCGGCCGGG GCGTGCCCGA GGGGAGGGAC
TCCCCGGCTT GCGACCCGGC GTTGTCCGCG
GAGGGCGCGC ACGAGGTCGA TCTTTCTGCG
TCTGGCGGAA AGGGTGAGCG CCCGAGGGCC ATGCCCGCGC
GCTTCAGGGC CGCGGACCCC GCGCGGCGCC TCCCGTCTCT
TTGGGCCGCC AAGATGAGCA AATACTGGGC GGTCGGGGGC
GGGACCAGGG CGACCCGGC GTTGTCCGCG
Disease genes identified
Sickle-cell anemia
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harmless misprints
Genetic polimorphisms variations over 1
frequency in humans
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Genetic background of memory loss
Genetic disease
Genetic susceptibility
Autosomal dominant Mutation cases (lt5)
PS 2 (0,1)
APP (0,4)
???
PS 1 (3)
Richard F, Amouyel P. Genetic susceptibility
factors for Alzheimer's disease. Eur J Pharmacol.
2001 412(1)1-12.
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Genetic background of memory loss
Genetic disease
Genetic susceptibility
apoE ?4 (Odds ratio 12.5)
Sporadic cases (95)
???
Richard F, Amouyel P. Genetic susceptibility
factors for Alzheimer's disease. Eur J Pharmacol.
2001 412(1)1-12.
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The candidate genotype
(2x - 10x)
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Genetic association study ofindividual
differences
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Allele frequencies can be ethnically different
http//www.Genomee.uci.edu/images/WorldMap_DRD4All
eles.jpg
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D Hamer and L Sirota Population stratification is
a potential source of error in psychiatric genetic
s. New study designs and statistical methods can
help guard against this problem. Molecular
Psychiatry (2000) 5, 1113.
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Genetic associations of the

• with personality traits

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Temperament
Character

Automatic responses to
Novelty
Danger
Behavior
Reward
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Searching for the genetic background of the
temperament dimensions
Novelty Seeking and the DRD4-VNTR 7-repeat
?
Ebstein, 1996 Nature Genetics Benjamin,
1996 Nature Genetics
Malhotra, 1996 Mol Psych Gelertner, 1997 Nature
Genetics Vandenbergh, Mol Psy (1997) Sander,
1997 Am J Med Genet Jonsson, 1998 Psych
Medicine Comings, 1999 Am J Med Genet
?
?
?
?
?
Ono, 1997 Med Genetics Ebstein, 1997 Mol
Psych Noble, 1998 Am J Med Genet Tomitaka,
1999 Am J Med Genet Stroebel, 1999 Mol Psych
?
?
?
?
?
?
?
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Based on Cloningers psycho-biological model,
using the Temperament and Character Inventory
(TCI)
157 unrelated, ethnically homogeneous Caucasians
(Hungarians)
240 questions yes/no answer 1 / -1 points
Non-invasive DNA sampling!
Publications

• Ronai, Z., Szekely, A., Nemoda, Z., Lakatos, K,
  Gervai, J., Staub, M., Sasvari-Szekely, M. (2001)
  Association between novelty seeking and the
  -521c/t polymorphism in the promoter region of
  the DRD4 gene. Molecular Psychiatry, 2001, 6 (1),
  35-38.
• Szekely, A., Ronai, Z., Nemoda, Z., Gervai,
  J., Sasvari-Szekely, M. (2003) Association
  between the long allele of the dopamin d4
  receptor gene and the persistence personality
  trait. American Journal of Medical Genetics Part
  B-Neuropsychiatric Genetics, (in press).

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Cheek-swab collection, storage and mailing
procedures

• swab inner cheek (20sec)
• place swab at purple area (5sec)
• let dry (2-3 hours turns white)
• store at room temperature in kit-bag
• mail for DNA analysis

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Cheek-swab collection, storage and mailing
procedures
Item 10534172
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phenotype
Validation of the methods for the

• Character dimensions of personality did NOT show
  genetic association!

• The reliability measures for the Hungarian
  version were satisfactory in each dimension!

• There were NO significant differences of sex in
  persistence!

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genotype
Validation of the methods for the

• The genotype and allele frequencies of the sexes
  are the same!

• The genotype frequencies are in Hardy-Weinberg
  equilibrium!

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RESULTS Association of the and the PERSISTENCE
temperament scale
4,4 4,7
4,4 4,7
Male individuals with the 7-repeat allele are
LESS PERSISTENT!!!
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Association studies between ADHD and the DRD4
VNTR 7-repeat
case-control
yes
no
TDT
meta-analysis Faraone, 2001 and Maher, 2002
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Junior Temperament and Character Inventory
(JTCI) in children with Attention Deficit
Hyperactivity Syndrome
167 unrelated children with ADHD ethnically
homogeneous Caucasians (Hungarians)
82 questions yes/no answer 1 / -1 points
Manuscript

• Tarnok, Z., Kiraly, O., Szantai, E., Szekely,
  A., Bognar, E., Gadoros, J., Sasvari-Szekely M.,
  and Nemoda Z. (manuscript) Association of
  dopamine d4 receptor polymorphisms with
  personality traits in a child psychiatric sample.

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RESULTS Association of the and the PERSISTENCE
temperament scale
LESS PERSISTENT!!!
ADHD children with the 7-repeat allele are
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RESULTS interaction between the DRD4-7repeat and
the -521 CT genotypes in predicting persistence
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Genetic associations of the

• with sustained attention

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The 7-repeat allele cognitive performance in
tasks that demand sustained attention
Task 1 Word Reading
Task 2 Picture Naming
7 repeat present slower reaction time (plt0.05)
7 repeat present slower reaction time (plt0.05)

• several hundred stimuli items
• 40-50 minutes sessions

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The mean reaction time performance of the 7
repeat group is slower
Task 1 Word Reading
Task 2 Picture Naming
Task 3 Word Repetition
Based on the collapsed reaction time data of 92
subjects on the two cognitive tasks the 7-repeat
allele group was 0.5 SD slower (plt0.01)
Szekely, Ronai, Nemoda, Gervai,
Sasvari-Szekely Association of the dopamine d4
receptor gene polimorphism and reaction time
performance in normal subjects
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More robust, or earlier fatigue effects?
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Summary and future directions

• individuals with the 7-repeat DRD4 VNTR are
  generally slower in a cognitive task that demands
  sustained attention

• males of this genotype can also be characterized
  as less persistent on a self-report questionnaire

• age?

• language and culture?

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Future directions Gene-by-environment interaction
Influence of Life Stress on Depression
Moderation by a Polymorphism in the 5-HTT Gene
Caspi, A., Sugden, K., Moffitt, T.E., Taylor, A.,
Craig, I.W., Harrington, H.L., McClay, J., Mill,
J., Martin, J., Braithwaite, A., and Poulton, R.
Science Jul 18 2003 386-389.
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Genetic Association Study of the Dopamine D4
receptor gene polimorphism and REACTION TIME
PERFORMANCE in normal subjects
Anna Szekely anna_at_crl.ucsd.edu Dr. Zsolt
Ronai, Dr. Zsofia Nemoda Dr. Maria Sasvari-Szekely