RESEARCH ARTICLE

Neuropsychiatric Genetics

Cadherin-13 Gene is Associated with

Hyperactive/Impulsive Symptoms in
Attention/Deficit Hyperactivity Disorder
Angelica Salatino-Oliveira,1 Julia Pasqualini Genro,1 Guilherme Polanczyk,2,3 Cristian Zeni,5
Marcelo Schmitz,4,5 Christian Kieling,5 Luciana Anselmi,6 Ana Maria Baptista Menezes,6
Fernando Cde Barros,6 Evelise Regina Polina,1 Nina R Mota,1,7 Eugenio Horacio Grevet,4,7
Claiton Henrique Dotto Bau,1,7 Luis Augusto Rohde,3,4 and Mara Helena Hutz1*
1

Department of Genetics, Universidade Federal do Rio Grande do Sul, RS, Brazil

Department of Psychiatry, Universidade de Sao Paulo, SP, Brazil

2
3

Institute for Developmental Psychiatry for Children and Adolescents, SP, Brazil

Department of Psychiatry, Universidade Federal do Rio Grande do Sul, RS, Brazil

Division of Child and Adolescent Psychiatry, Hospital de Clnicas de Porto Alegre, RS, Brazil

5
6

Postgraduate Program in Epidemiology, Universidade Federal de Pelotas, RS, Brazil

Adult ADHD Outpatient Program, Hospital de Clnicas de Porto Alegre, RS, Brazil

Manuscript Received: 8 September 2014; Manuscript Accepted: 10 December 2014

Several efforts have been made to find new genetic risk variants
which explain the high heritability of ADHD. At the genome
level, genes involved in neurodevelopmental pathways were
pointed as candidates. CDH13 and CTNNA2 genes are within
GWAS top hits in ADHD and there are emerging notions about
their contribution to ADHD pathophysiology. The main goal of
this study is to test the association between SNPs in CDH13 and
CTNNA2 genes and ADHD across the life cycle in subjects with
ADHD. This study included 1,136 unrelated ADHD cases and
946 individuals without ADHD. No significant association between CDH13 and CTNNA2 was observed between cases and
controls across different samples (P
0.096 for all comparisons).
No allele was significantly more transmitted than expected
from parents to ADHD probands. The CDH13 rs11150556 CC

How to Cite this Article:

Salatino-Oliveira A,Genro JP, Polanczyk G,
Zeni C, Schmitz M, Kieling C, Anselmi L,
Menezes AMB, Barros FC, Polina ER,
Mota NR, Grevet EH, Bau CHD, Rohde
LA, Hutz MH. 2015. Cadherin-13 Gene is
Associated with Hyperactive/Impulsive
Symptoms in Attention/Deficit
Hyperactivity Disorder.
Am J Med Genet Part B 168B:162169.

Conflict of Interest: Dr. Luis Augusto Rohde has been a member of the speakers bureau/advisory board and/or acted as a consultant for Eli-Lilly,
Janssen-Cilag, Novartis, and Shire in the last three years. He receives authorship royalties from Oxford Press and ArtMed. He has also received travel
awards from Shire for his participation of the 2014 APA meeting. The ADHD and Juvenile Bipolar Disorder Outpatient Programs chaired by him
received unrestricted educational and research support from the following pharmaceutical companies in the last three years: Eli-Lilly, Janssen-Cilag,
Novartis, and Shire. He also receives research support from Brazilian government institutions (CNPq, FAPERGS, HCPA, and CAPES). Dr. Guilherme
Polanczyk receives research support from the National Council for Scientific and Technological Development (CNPq), the Sao Paulo Research
Foundation (FAPESP), and the University of Sao Paulo. He has served as a consultant to Shire. He has served on the speakers bureau of Shire. He has
received royalties from Editora Manole. Dr Mara H. Hutz receives support from Brazilian government institutions (CNPq, FINEP, and CAPES). Dr
Anselmi and Dr Kieling receive governmental CAPES/FAPERGS post-doctoral fellowships. In the last 36 months, Dr Kieling has received royalties
from publishers Artmed and Manole, as well as financial support from Brazilian governmental funding agencies (FIPE/HCPA, CNPq, CAPES, and
FAPERGS). Dr Grevet received support from Shire in the last three years. None of the other authors have any conflicts of interest to declare.

Correspondence to:
Prof. Mara H. Hutz, Departamento de Genetica, Instituto de Biociencias, UFRGS Caixa PO Box No. 15053, CEP Porto Alegre, RS, Brazil 91501-970.
E-mail: [email protected]
Article first published online in Wiley Online Library (wileyonlinelibrary.com)
DOI 10.1002/ajmg.b.32293

2015 Wiley Periodicals, Inc.

162

SALATINO-OLIVEIRA ET AL.
genotype was associated with more hyperactive/impulsive symptoms in youths with ADHD (children/adolescents clinical sample: F 7.666, P 0.006, FDR P-value 0.032; Pelotas Birth
Cohort sample: F 6.711, P 0.011, FDR P-value 0.032).
Although there are many open questions regarding the role of
neurodevelopmental genes in ADHD symptoms, the present
study suggests that CDH13 is associated with hyperactive/
impulsive symptoms in youths with ADHD.
2015 Wiley Periodicals, Inc.

Key words: neurodevelopment; CDH13; CTNNA2; cadherins;

ADHD

INTRODUCTION
Attention-deficit/hyperactivity disorder (ADHD) is a common
neurodevelopmental disorder, as defined by the Diagnostic and
Statistical Manual of Mental Disorders - fifth edition (DSM-5,
American Psychiatry Association, 2013). The ADHD is characterized by a pattern of age-inappropriate behavior symptoms divided
into two dimensions: inattention and hyperactivity/impulsivity.
Although the diagnostic criteria distinguish three different presentations of ADHD according to predominant symptoms, the
disorder can be better understood dimensionally [Hudziak, 2007;
Kraemer, 2007; Salum et al., 2014]. This disorder is one of the most
prevalent mental health disorders in childhood and adolescence,
affecting 5.29% of children worldwide [Polanczyk et al., 2007]. Even
though the proportion of persistence and prevalence rates of ADHD
in adults are still controversial [Matte et al., 2014], there are studies
suggesting that up to 66% of ADHD children will have clinically
significant symptoms of the disorder in adulthood [Barkley et al.,
2002; Haavik et al., 2010].
ADHD is a very heterogeneous disorder and its etiology remains
unclear. However, several studies support a strong genetic contribution. A meta-analysis of twin studies estimated an average ADHD
heritability of 76% [Faraone et al., 2005]. Although ADHD is highly
heritable, few variants with small effect were identified that explain a
small portion of the estimated heritability. Several genome-wide
association (GWAS) studies have been conducted. Despite without
statistically significant results at the genome level, genes related to
neurotransmission and cell-cell communication systems were
suggested, including genes related to processes such as cell division,
adhesion and polarity, neuronal migration and plasticity, extracellular matrix regulation, and cytoskeletal remodeling [Poelmans
et al., 2011]. These findings from genome-wide approaches indicate
a whole range of new and promising possibilities for ADHD
molecular genetic studies [Akutagava-Martins et al., 2013]. A
pathway-based analysis using case-only design showed that neurite
outgrowth genes pointed by Poelmans et al. (2011) were associated
with hyperactive/impulsive scores in ADHD children from the
International Multicenter ADHD Genetics (IMAGE) study [Bralten et al., 2013].
The most promising gene derived from these studies is CDH13
which was among top hits of three independent investigations
[Lasky-Su et al., 2008; Lesch et al., 2008; Neale et al., 2010]. This gene

163
is located at chromosome 16q23.3 and encodes cadherin-13 which
belongs to a large family of transmembrane adhesion calciumdependent molecules [Philippova et al., 2009]. However, the cadherin-13 protein is an atypical cadherin. It lacks transmembrane
and cytosolic domains and it is attached to the cellular membrane
by a glycosylphosphatidylinositol (GPI) anchor [Philippova et al.,
2009]. This characteristic of cadherin-13 allows this protein to have
a critical role as a signaling molecule differently from classical
cadherins [Rivero et al., 2013]. The CDH13 transcripts were found
in prefrontal cortex (PFC), medulla, thalamus, and midbrain of the
human adult brain whereas the protein was detected in cerebral
cortex, medulla oblongata and nucleus olivaris cellular membranes
and neurites [Takeuchi et al., 2000]. The CDH13 functions could be
potentially related to neurite outgrowth, dendrite arborizations,
synapse development, and maintenance of synaptic contacts [Rivero et al., 2013].
Two candidate gene studies in independent samples were performed with CDH13 and ADHD. The first tested the association
between this gene and three executive function tasks. The
rs11150556 polymorphism was associated with verbal workingmemory (VWM) performance in children with ADHD. Carriers of
the CC genotype showed a significant worse performance when
compared to CT and TT carriers [Arias-Vasquez et al., 2011]. The
second study aimed to identify, characterize and estimate coding
CDH13 variants in adults with ADHD [Mavroconstanti et al.,
2013]. Seven coding variants were found; only one was novel
and none of them showed significant associations with ADHD.
Moreover, no significant differences were observed between
expression levels of these coding variants in either HEK293 or
CHO cells [Mavroconstanti et al., 2013].
Another gene involved in neurodevelopmental pathways that
figures at the GWAS top hits is the CTNNA2 [Poelmans et al.,
2011]. This gene encodes the aN-catenin protein and is located at
chromosome 2p12-p11.1 [Kobielak and Fuchs, 2004]. The
a-catenins binds typical cadherins with the actin cytoskeleton
and it maintains the stability of dendritic spines and synaptic
contact [Abe et al., 2004]. The aN-catenin is highly expressed in
the central nervous system, particularly in hypothalamus, amygdala, cingulate cortex, temporal lobe, and PFC [Terracciano et al.,
2011]. Adhesion junctions in border active zones in developing and
mature synapses have active aN-catenin throughout the developing
and postnatal brain [Terracciano et al., 2011]. aN-catenins seems to
stabilize synapse formation mediated by typical cadherins during
the development of nervous system. Further studies have supported
the involvement of CTNNA2 in ADHD etiology. The CTNNA2
gene is located within CNVs in individuals with ADHD [Elia et al.,
2010]. Moreover, a GWAS identified an association between scores
in the excitement-seeking scale of the revised Neuroticism-Extraversion-Openness personality inventory and CTNNA2 gene. This
trait is a main component of impulsivity and it is a core feature of
extraversion [Terracciano et al., 2011]. This result suggests a role of
the CTNNA2 gene in the biological basis of the excitement-seeking
and impulsive behaviors that are common in patients with ADHD
[Terracciano et al., 2011].
Therefore, there are emerging data about the CDH13 and
CTNNA2 contribution to the pathophysiology of ADHD.
Candidate gene studies in independent samples are required in

164
order to validate GWAS top-findings and to determine the role of
these genes in ADHD etiology. The main goal of this study was to
test the association between single nucleotide polymorphisms
(SNPs) in CDH13 and CTNNA2 genes and ADHD across the
life cycle in subjects with ADHD.

MATERIAL AND METHODS

Subjects
This study included a total of 1,136 unrelated ADHD cases and 946
individuals without ADHD who came from three sources where the
phenotype was extensively assessed. The first sample consisted of
526 ADHD children and adolescents and their biological parents.
These patients were recruited at the ADHD Outpatient Program
(ProDAH) from Hospital de Clnicas de Porto Alegre. The sample
was diagnosed according to the Diagnostic and Statistical Manual
of Mental Disorders (DSM-IV) criteria [American Psychiatric
Association, 1994], following a previously reported three-stage
protocol [Roman et al., 2001], including the application of a
semi-structured diagnostic interviews (KSADS-PL) and clinical
assessments by experienced child psychiatrists. The Swanson,
Nolan, and Pelham Scale-Version IV (SNAP-IV) was applied to
277 of those patients by child psychiatrists blinded to genotype. A
control group of 129 children and adolescents without evidence of
any current or past mental illness as defined by the DSM-IV was
selected. The diagnostic approach for the control sample was the
same as for the ADHD samples [Schmitz et al., 2006].
The second ADHD sample comprises 110 individuals from the
1993 Pelotas Birth Cohort. These subjects were evaluated at 1819
years of age in a one-stage procedure. A structured interview based
on the Diagnostic and Statistical Manual of Mental Disorders
5th edition (DSM-5) was performed [American Psychiatric
Association, 2013]. The ADHD in adults in DSM-5 is defined as
the presence of at least five among nine symptoms of inattention
and/or five among nine symptoms of hyperactivity/impulsivity.
ADHD symptoms must cause clinical interference, several of them
must be present in more than one setting, and their age of onset
should be before 12 years of age. The DSM-5 ADHD symptoms
were rated as present or absent. Initially, a screening questionnaire
using the same structure of the six-question World Health Organization Adult ADHD Self-Report Scale Screener (ASRS) was applied
for all subjects. This screening consists of six questions about
ADHD symptoms (two hyperactivity items: on the go, and
fidgets; and four inattention items: forgetful, does not follow
through, reluctant to engage in mental tasks, and difficulty
organizing tasks), which were adapted to the DSM-5 wording.
Considering a cut-off of 4/6 screening symptoms, ASRS had 68.7%
sensitivity, 99.8% specificity, and 97.9% total classification accuracy in a previous population study [Kessler et al., 2005]. In order to
enhance sensitivity, any subject with two or more positive questions
among the six was considered screening positive in our study. In this
case, they answered questions about the 12 remaining ADHD
symptoms, as well as about additional criteria (clinical impairment,
symptom pervasiveness, and age of onset before 12 years old).
Pervasiveness was assessed by questioning if the subject presented
symptoms in at least two of the three main settings: home, social,

AMERICAN JOURNAL OF MEDICAL GENETICS PART B

and work/school environments. To measure the clinical
impairment specifically related to ADHD, a scale ranging from 0
(no impairment) to 3 (severe impairment) was applied at the end of
the ADHD assessment interview. Clinical impairment was defined
as scores of 2 (moderate) or 3 (severe). A total of 508 individuals
were selected from the 1993 Pelotas Birth Cohort as controls. This
control sample includes all subjects with positive screening (i.e., at
least two positive questions in the six question screening) who did
not meet full ADHD diagnostic criteria in the subsequent evaluation (n 179) and individuals with none of the six ADHD screening symptoms (n 331; for more details see [Matte et al., 2014]).
Methodology of data collection and demographic data from this
Cohort are described fully in Victora et al. (2006) and Matte et al.
(2014). The Mini International Neuropsychiatric Interview (MINI)
was performed to evaluate several other psychiatric diagnoses. The
MINI is a short semi-structured diagnostic interview for DSM-IV
and ICD-10 psychiatric disorders, which provides prevalence estimates of common mental disorders. Only some MINI sections were
used due to logistic issues (i.e, the psychiatric interview was part of a
larger follow-up assessment). The most prevalent mood and anxiety
disorders were assessed: bipolar, major depression, agoraphobia,
generalized anxiety, and social phobia. The MINI has a previously
validated Portuguese version [Amorim, 2003]. The MINI had
kappas of 0.650.85, sensitivity of 0.750.92, and specificity of
0.900.99 when using Structured Clinical Interview for Diagnosis
(SCID) applied by a psychiatrist as a parameter in primary health
care in Brazil [de Azevedo Marques and Zuardi, 2008].
The third ADHD sample comprises 500 adults with ADHD that
were also recruited at ProDAH. The diagnostic procedures for
ADHD and comorbidities followed the DSM-IV criteria. The
ADHD and oppositional defiant disorder diagnoses followed the
same three-staged procedure described above for children and
adolescents. Questions from K-SADS-E designed for children
were adapted for adults [Grevet et al., 2005]. Axis I psychiatric
comorbidities were evaluated using the Structured Clinical Interview for DSM-IV, research version (SCID-I-RV) [First et al., 1998].
The diagnoses of conduct disorder and anti-social personality
disorder were obtained using the appropriate sections of the
Mini International Neuropsychiatric Interview (MINI) [Amorim,
2003]. The adult control group of 309 individuals was evaluated in a
blood donation center in the same hospital. The inclusion criteria
were: (i) be Brazilian of European descent; and (ii) 18 years of age or
older. The exclusion criteria were the same as those used for the
ADHD patients (mentioned above), as well as the fulfillment of a
DSM-IV ADHD diagnosis. Furthermore, the SNAP-IV questionnaire was also applied.
This study was approved by the Ethics Committee of the Universidade Federal de Pelotas and by the Ethics Committee of
Hospital de Clnicas de Porto Alegre. Adults were invited to
participate and provided a written informed consent. For children,
the parents provided written informed consent, and the children/
adolescents provided verbal assent for participation in this study.

Genotyping
Blood samples were collected from patients enrolled at ProDAH
and their parents whenever possible. DNA was extracted from

SALATINO-OLIVEIRA ET AL.

Case
33.6  11.1
265 (53.0%)
500 (100%)
57 (18.4%)
266 (53.2%)
203 (40.6%)
P-valueb

0.205
0.168
<0.001

<0.001
c

Data are given as number (percentage) or mean ( standard deviation).

Calculated by the MannWhitney U-test (quantitative variables without normal distributions), the x2-test (categorical variables).
Agoraphobia, generalized anxiety disorder, social phobia.

Control
18
228(44.5%)
353(69.5%)
55(10.8%)

13(2.6%)

1993 Pelotas birth cohort

Case
18
42 (38.2%)
69 (62.8%)
55 (50.0%)

23 (20.9%)
P-valueb
0.001
0.001
0.269
0.410
<0.001
<0.001
Control
11.6  3.14
80(62.0%)
98(76.0%)
25(19.4%)
14(10.9%)
2(1.6%)
Case
10.5  3.08
403 (76.6%)
409 (77.7%)
120 (22.8%)
260 (49.4%)
80 (15.2%)

Children with ADHD were younger and predominantly males

when compared to controls whereas the adults with ADHD were
older than their controls. Most individuals are of European ancestry, although some individuals of African ancestry were also included. The most common comorbid disorders observed in these
ADHD patients were anxiety, disruptive behavior, and mood
disorders (Table I).
Allele frequencies were estimated for each SNP within each
group. Genotype distributions were in Hardy-Weinberg equilibrium in all samples (Table SI). The D among SNPs at CDH13 gene
was D 0.009 between rs11646411 and rs6565113, D 0.062
between rs11646411 and rs11150556, and D 0.010 between
rs6565113 and rs11150556. These D values indicated absence of
LD among these polymorphisms.

Characteristics
Age (years)
Gender (male)
Ethnicity (EuropeanBrazilian)
Anxiety disordersc
Disruptive behavior disorders
Mood disorders

RESULTS

Children/Adolescents from ProDAH

Allele frequencies were estimated by counting. Deviations from

HardyWeinberg equilibrium were assessed by the x2 test. The
PLINK program v.1.07 was used to estimate the D as a measure of
pairwise linkage disequilibrium (LD) between SNPs at CDH13 gene
[Purcell et al., 2007]. Comparisons among variables were performed using the x2, Fishers exact or MannWhitney U-test
(for quantitative variables without normal distributions). Potential
confounders to be entered in the models were defined based on
conceptual analyses of the literature and by means of a statistical
definition (association with the study factor and with the outcome
at P < 0.10). The association of the SNPs with ADHD susceptibility
was tested in a casecontrol design by multivariate logistic regression analysis in each group. Hyperactivity/impulsivity SNAP-IV
scores were compared among genotypes by ANOVA. The effect size
for significant P-values was obtained through partial eta squared.
All these tests were performed with SPSS Version 18 software (SPSS,
Inc., Chicago, IL). The family-based association tests were carried
out with the UNPHASED v.3.1.7. software [Dudbridge, 2008]. The
significance level accepted for all tests was 0.05 and the adjustment
for multiple testing by false discovery rate (FDR) procedure was
performed [Benjamini and Hochberg, 1995].

TABLE I. Demographic and Clinical Characteristics of Each Groupa

Statistical Analyses

Adults from ProDAH

lymphocytes by standard procedures. DNA samples from the 1993

Pelotas Birth Cohort were obtained from saliva, using Oragene1
OG-250 DNA Self-Collection kit, following the manufacturers
recommended protocol (DNA Genotek Inc., Kanata, Ontario,
CA). DNA from all samples was quantified by spectrophotometry
using NanoDrop 1000 (Thermo Fisher Scientific Inc., Waltham,
MA) and diluted to a standard concentration of 10 ng/mL. The
polymorphisms rs11646411, rs6565113, and rs11150556 in CDH13
and rs13395022 in CTNNA2 genes were genotyped using TaqMan
SNP Genotyping Assays-on-Demand (Applied Biosystems, Foster
City, CA) according to the manufacturers recommended protocol.
Three SNPS (CDH13 rs11646411, rs6565113) and CTNNA2
rs13395022) were chosen because they are GWAS top hits [Poelmans et al., 2011]. CDH13 rs11150556 was included due to its
association with verbal working-memory in children with ADHD
[Arias-Vasquez et al., 2011].

Control
30.2  9.1
143(46.3%)
309(100%)
159(31.8%)
6(2.0%)
80(25.9%)

P-valueb
<0.001
0.071

<0.001
<0.001
<0.001

165

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AMERICAN JOURNAL OF MEDICAL GENETICS PART B

The distribution of ADHD among SNP genotypes in CDH13 and

CTNNA2 genes was analyzed by logistic regression in each
case-control sample. No significant association was observed in
case-control analyses across the three different samples (Table II).
The association of the rs11150556 CC genotype with ADHD in the
1993 Pelotas birth cohort was not maintained after FDR correction
(F 1.601, P-value 0.032, FDR P-value 0.096; Table II). Family-based association tests for each investigated variant in the
children/adolescents ADHD sample were performed. No allele
was significantly more transmitted than expected from parents
to ADHD probands (P
0.211 for all SNPs; data not shown, but
available upon request).
Significant differences in hyperactivity/impulsivity scores in the
SNAP-IV between CDH13 rs11150556 genotype groups in children/adolescents with ADHD (F 3.901, P 0.011, FDR P-value
0.026; Table III) were observed. Similar differences were found in
DSM-5 hyperactivity/impulsivity symptom count in ADHD individuals from the 1993 Pelotas birth cohort (F 4.116, P 0.022,
FDR P-value 0.033; Table III). When CDH13 CC carriers were
compared to CT TT carriers, the difference was more evident.
The CDH13 CC genotype was associated with higher hyperactive/
impulsive symptoms in children/adolescents and young people
with ADHD (children/adolescents: F 7.666, P 0.003, FDR
P-value 0.018; Pelotas Birth Cohort: F 6.711, P 0.013, FDR
P-value 0.026; Table III). The corresponding effect sizes were 3%
and 7% for children/adolescents and Pelotas birth cohort samples,
respectively. This association was not observed in adult patients.

DISCUSSION
The selected polymorphisms at the CDH13 and CTNNA2 genes
were not associated with ADHD through case-control analyses in all

age-different samples. Moreover, family-based tests indicated that

there was no over-transmission of any allele of these variants from
parents to children/adolescents with ADHD. However, significantdifferences in hyperactive/impulsive symptoms in children/adolescents
and young people with ADHD according to the CDH13 rs11150556
genotypes were observed. These results suggest that CDH13 rs11150556
CC genotype could lead to increased hyperactive/impulsive symptoms
in the early period of the life cycle of ADHD individuals.
Our findings evoke three interesting issues: (i) what would be the
reasons for positive findings in hyperactive scores only during
childhood and adolescence/young adulthood? (ii) Why only positive findings in dimensional scores and not in categorical diagnosis?
(iii) Why positive findings in hyperactive/impulsive dimension and
not in inattentive scores?
Although ADHD in children and adolescents often persists into
adulthood, the symptomatic presentation of the disorder at later
age phases differs from that in children [Volkow and Swanson,
2013]. A fraction of this difference is due to a greater decrease in
hyperactive/impulsive symptoms than in inattentive symptoms
[Matte et al., 2012; Willcutt et al., 2012; Volkow and Swanson,
2013]. The developmental instability of ADHD symptoms across
the life cycle could be reflected by the diagnostic changes in DSM-5,
which eliminated the previous enumeration of three subtypes of
ADHD [Volkow and Swanson, 2013]. Genetic and environment
contributions to the developmental course and outcomes of ADHD
in adulthood are relatively understudied and more investigations
are needed [Franke et al., 2012]. Interestingly, even though a high
heritability of ADHD in children is clearly found, some studies
estimate a low heritability for ADHD symptoms in adults [Franke
et al., 2012] suggesting different etiologic pathways for different
ages ranges. Furthermore, the cortical thickness trajectories seem to
differ according to ADHD remission or persistence into adulthood

TABLE II. Logistic Regression of the SNPs in CDH13 and CTNNA2 Genes With presence of ADHD Across the Three Groups
Children/adolescents
from ProDAHa
SNP
rs11646411

Gene
CDH13

Genotypes
G_b
CC

rs6565113

CDH13

TT
GT
GG

rs11150556

CDH13

T_c
CC

rs13395022

CTNNA2

C_b
TT

OR (95% CI)
1
0.968
(0.6041.550)
1
0.903
(0.5711.428)
0.872
(0.5021.515)
1
1001
(0.6481.545)
1
1.185
(0.7881.782)

P-value

0.892

0.663
0.626

0.999

0.415

1993 Pelotas birth cohort

OR (95%CI)
1
1.465
(0.8582.502)
1
1.018
(0.6411.615)
0.892
(0.4661.706)
1
1.601
(1.0402.465)
1
1.098
(0.7201.674)

P-value

0.162

0.940
0.730

0.096

0.664

Adults from ProDAHa

OR (95%CI)
1
0.938
(0.6601.334)
1
1.188
(0.8431.674)
0.785
(0.5211.184)
1
1.032
(0.7441.431)
1
1.260
(0.9351.697)

SNP, Single Nucleotide Polymorphism; OR, Odds Ratio; CI, Confidence Interval.
a
Multivariate logistic regression adjusted for gender and age.
b
The homozygous for the minor allele and the heterozygous genotypes were polled due to low frequency.
c
T carriers were pooled with CC homozygous because they were previously associated with worse performance in VWM task; P-values were corrected by false discovery rate.

P-value

0.723

0.325
0.248

0.999

0.128

SALATINO-OLIVEIRA ET AL.

167

TABLE III. Mean Hyperactivity/Impulsivity Symptoms According to rs11150556 CDH13 Genotypes Across the Three Groups
95% IC
Sample
Children/adolescents
from ProDAHa

1993 Pelotas birth

cohortb

Adults from ProDAHc

rs11150556
genotypes
TT
CT
CC

N
52
144
80

Mean
1.521
1.443
1.771

 SE
0.127
0.087
0.104

Lower
bound
1.271
1.273
1.567

Upper
bound
1.771
1.614
1.976

T_d
CC
TT
CT
CC

196
80
22
46
42

1.461
1.768
4.10
4.74
5.54

0.082
0.104
0.426
0.305
0.320

1.300
1.564
3.25
4.13
4.90

T_d
CC
TT
CT
CC
T_d
CC

68
42
108
243
131
351
131

4.52
5.53
1.518
1.518
1.453
1.518
1.453

0.250
0.321
0.071
0.048
0.065
0.040
0.065

4.02
4.89
1.378
1.424
1.326
1.440
1.326

P-value

FDRe

4.579

0.011

0.026

1.622
1.972
4.94
5.34
6.17

8.800

0.003

0.018

3.969

0.022

0.033

5.02
6.16
1.658
1.612
1.580
1.596
1.580

6.400

0.013

0.026

0.370

0.691

0.691

0.742

0.389

0.456

Hyperactive/Impulsive SNAP-IV scores was the dependent variable; ANOVA was adjusted for gender and ethnicity.
The dependent variable in the models was DSM-V hyperactivity/impulsivity symptoms; ANOVA was adjusted for ethnicity.
Hyperactive/Impulsive SNAP-IV scores was the dependent variable; ANOVA was performed with adjustment for gender.
d
TT CT against the CC genotype.
e
P-values adjusted for false discovery rate correction.
b
c

from childhood [Shaw et al., 2013]. Comparing to typically developing subjects, persistent ADHD patients showed a fixed deficit in
cortical maturation whereas remitted ADHD individuals converted
to normal maturation pattern. Moreover, the link between cortical
trajectories and ADHD outcome seems to be driven by inattentive
symptoms [Shaw et al., 2013].
The majority of Cadherin-13 expression studies are only suggestive regarding the involvement of this atypical cadherin in the
regulation of brain development and function. Several experimental findings indicate that Cadherin-13 expression is not restricted to
a specific cell type suggesting a key role for Cadherin-13 in several
basic functions of the developing and adult brain [Rivero et al.,
2013]. Moreover, Cadherin-13 expression pattern overlaps with
regions showing volumetric reductions in patients with ADHD,
such as the cerebellum and pre-frontal cortex [Valera et al., 2007];
[Konrad and Eickhoff, 2010].
During mammalian brain development, monoaminergic terminals coming from the dopaminergic substantia nigra and ventral
tegmental area, the noradrenergic locus coeruleus or the serotonergic
dorsal raphe will be guided to their peripheral prefrontal cortical
targets by different membrane bound or soluble proteins that act as
guidance molecules. One of these guidance molecules would be
Cadherin-13, with a critical function as a signaling molecule rather
than a typical adhesion molecule [Riveros et al, 2013]. Although
different lines of evidence suggest Cadherin-13 to be a negative
regulator of neurite extension and subsequent axonal pathfinding,
the precise mechanism of Cadherin-13 action is unknown, and the

role of Cadherin-13 in synaptogenesis remains elusive. Cadherin-13

expression appears to be higher in the adult than in the developing
brain, at least in humans [Takeuchi et al., 2000]. Therefore a certain
versatility of Cadherin-13 throughout the lifespan seems most likely,
first as an axonal pathfinder during neurodevelopment, and, once
the neuronal circuits have been established, it may be more relevant
for the maintenance of synapses and/or the coordination of processes critical for synaptic plasticity [Riveros et al., 2013].
Symptomatic threshold for diagnosing ADHD is a controversial issue and the disorder seems to be better understood at
the dimensional level [Larsson et al., 2012; Salum et al., 2014]. A
quantitative view of ADHD could facilitate etiologic investigations, considering that genetic and environmental factors
seem to operate dimensionally throughout the distribution
of ADHD symptoms [Larsson et al., 2012]. Moreover, genetic
studies of symptom domains might increase the strength of
association analysis due to a decrease in phenotypic heterogeneity [Bralten et al., 2013]. Therefore, in addition to genetic
association analyzes by case-control strategies, quantitative
analyses should be performed in genetic studies of some complex phenotypes, such ADHD, when it is possible or plausible
[Lewis and Knight, 2012].
Considering the decrease in hyperactive/impulsive symptoms
over the life cycle, association analysis of this symptom domain with
neurodevelopmental genes at different age ranges seems to be
relevant to understand ADHD [Bralten et al., 2013]. Moreover,
genetic investigations of symptom domains separately might help

168
in genetic risk variants identification of specific dimensions [Bralten et al., 2013]. In both childhood and adulthood, a 6070%
genetic correlation between hyperactivity-impulsivity and inattention has been estimated in ADHD patients [Franke et al., 2012].
Therefore, even considering that these two symptom domains
compose ADHD diagnosis and share a proportion of genetic effects,
there is an important genetic specificity for each symptom group
[Greven et al., 2011].
This work should be viewed in light of certain limitations. First,
in this study there is a disproportion between cases and controls in
the children/adolescent and cohort samples. Ideally, a similar
number of case and control subjects would be expected. Indeed,
this disproportion between case and control subsamples tends to
increase the probability of a false positive finding. However, the
results of case-control analyses were negative [Balding, 2006].
Second, this is a cross-sectional study whereas it would be better
to investigate the involvement of neurodevelopmental genes in a
longitudinal approach. Third, although we included ethnicity as a
covariate in the ANOVA analyses, no genomic control was performed. Therefore, our findings could have been biased by hidden
genetic heterogeneity present in this sample. Putting all together,
these evidences suggest a putative role of CDH13 gene underlying
hyperactive/impulsive symptoms of ADHD during development.
Although the effect sizes were small, as would be expected for single
genes in multifactorial diseases, our results showed that the involvement of CDH13 in hyperactive/impulsive domain in ADHD
seems to be more evident in childhood and adolescence than in the
adulthood.

ACKNOWLEDGMENTS
The authors thank Conselho Nacional de Desenvolvimento Cientfico e Tecnologico (CNPq, Brazil), Coordenacao de Aperfeicoamento de Pessoal de Nvel Superior (CAPES, Brazil), and Fundo de
Incentivo a` Pesquisa e Eventos - Hospital de Clnicas de Porto
Alegre (FIPE/HCPA, Brazil) for financial support. The Welcome
Trust supports The Pelotas Birth Cohort Study. The European
Union and the Brazilian National Program for Centers of Excellence
(PRONEX CNPq) funded the initial phases of the 1993 Pelotas
Birth Cohort. Thanks are due to the patients that participated in the
study and the clinical staff of PRODAH-HCPA.

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