Progress in Neuro-Psychopharmacology & Biological Psychiatry 50 (2014) 178–183

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Progress in Neuro-Psychopharmacology & Biological

Psychiatry
journal homepage: www.elsevier.com/locate/pnp

Catechol-O-methyltransferase Val158Met polymorphism and altered

COMT gene expression in the prefrontal cortex of suicide brains
Lisheng Du a,⁎, Zul Merali a, Michael O. Poulter b, Miklós Palkovits c, Gábor Faludi d, Hymie Anisman e
a
University of Ottawa Institute of Mental Health Research, Ottawa, Ontario, Canada
b
Molecular Brain Research Group, Robarts Research Institute, University of Western Ontario, London, Ontario, Canada
c
Neuromorphological and Neuroendocrine Research Laboratory, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary
d
Department of Clinical and Theoretical Mental Health, Semmelweis University, Budapest, Hungary
e
Institute of Neuroscience, Carleton University, Ottawa, Ontario, Canada

a r t i c l e i n f o a b s t r a c t

Article history: Catechol-O-methyltransferase (COMT) plays a key role in the degradation of catecholamine neurotransmitters
Received 2 October 2013 within the brain. A functional polymorphism COMT Val158Met has been associated with psychiatric disorders
Received in revised form 6 December 2013 including suicidal behavior. In the present study we examined whether this polymorphism was related to
Accepted 17 December 2013
COMT mRNA expression in frontal cortical regions, and whether the expression of COMT differed between
Available online 2 January 2014
depressed suicide victims and psychiatric healthy controls. The Val158Met polymorphism was determined by
Keywords:
polymerase chain reaction and restriction fragment length polymorphism (PCR–RFLP) analysis. The levels of
COMT polymorphism COMT mRNA expression in the frontopolar cortex (FPC; 29 suicides vs. 27 controls) and orbital frontal cortex
Depression (OFC; 19 suicides vs. 15 controls) were significantly increased among depressed individuals that died by suicide
mRNA expression relative to those of controls, being up-regulated by approximately 60% and 65% in the FPC and OFC, respectively.
Prefrontal cortex Furthermore, among individuals with the Met allele (Met/Met and Met/Val genotypes) who died by suicide
Suicide COMT mRNA expression was elevated relative to that of the nondepressed Met allele carriers. However,
significant differences were not detected between suicides (n = 49) and controls (n = 72) with respect to
the Val158Met genotypic distribution and allelic frequencies. These results are consistent with the perspective
that altered COMT mRNA expression in frontal cortical brain regions might contribute to suicide and/
or depression, further supporting the role of dysregulation of catecholaminergic pathway genes in the
pathophysiology of suicide behaviors.
© 2014 Elsevier Inc. All rights reserved.

1. Introduction is most prominent within the context of psychiatric illness, particularly

Suicide is a major public health concern, with approximately one
million people committing suicide world-wide each year (WHO, 2012).
Risk factors for suicidal behavior include, among other things, psychiatric
and medical illness, impulsivity, aggression, alcohol and drug abuse, as
well as family/social stress (Mann, 2003; Pandey, 2011). Of these, suicide
Abbreviations: ANCOVA, analysis of covariance; ANOVA, analysis of variance; BDNF,
brain derived neurotrophic factors; COMT, catechol-O-methyltransferase; CSF, cerebrospi-
nal fluid; DAT, dopamine transporter; DLPFC, dorsolateral prefrontal cortex; DSM-IV,
Diagnostic and Statistical Manual of Mental Disorders, 4th Edition; FGF-2, fibroblast
growth factor-2; FPC, frontopolar cortex; GABA, γ-aminobutyric acid; GAPDH, glyceralde-
hyde 3-phosphate dehydrogenase; 5-HT, serotonin; 5-HTT, serotonin transporter; 5-
HTTLPR, serotonin-transporter-linked polymorphic region; Met, methionine; OFC, orbital
frontal cortex; PFC, prefrontal cortex; PMI, postmortem interval; RIN, RNA integrity num-
ber; SNP, single-nucleotide polymorphism; Val, valine.
⁎ Corresponding author at: University of Ottawa Institute of Mental Health Research,
1145 Carling Ave., Ottawa, Ontario K1Z 7K4, Canada. Tel.: +1 613 722 6521x7052; fax:
+1 613 792 3935.
E-mail address: lishengdu@gmail.com (L. Du).
depressive disorders. Dissociating to what extent any given neurobiolog-
ical factor underlies depression versus suicide, however, is very difficult,
although suicide is often preceded by negative life events that might
interact with genetic factors (Roy, 2012).
Neurobiological studies of depression/suicide have revealed several
correlates of suicide/depression, including disturbances of serotonin
(5-HT) receptors and of the serotonin transporter (5-HTT) (Jasinska
et al., 2012; Snyder, 2011), growth factors, such as brain derived
neurotrophic factors (BDNF) and fibroblast growth factor-2 (FGF-2)
(Duman and Monteggia, 2006; Dwivedi et al., 2005; Jeon et al., 2012;
Karege et al., 2005; Turner et al., 2012), as well as hormones, such as
estrogen/progesterone (Baca-Garcia et al., (2010)), FSH (Kim et al.,
2013), and corticotropin-releasing hormone (Austin et al., 2003). In
addition, there has been evidence linking dysregulation of catechol-
aminergic pathways with the development of depression/suicide
(Pitchot et al., 2003; Ryding et al., 2008; Tsai et al., 2011). In this regard,
particular attention has focused on the relationship between catechol-
O-methyltransferase (COMT) and depression/suicide. Ordinarily, this
enzyme catalyzes the transfer of a methyl group to catecholamines (e.g.,
dopamine and norepinephrine) and thus degradation of catecholamine

0278-5846/$ – see front matter © 2014 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.pnpbp.2013.12.016
 L. Du et al. / Progress in Neuro-Psychopharmacology & Biological Psychiatry 50 (2014) 178–183
neurotransmitters, dopamine and norepinephrine. Although COMT is
expressed throughout the human brain (Hong et al., 1998), it plays a
major role in regulating dopamine levels in the prefrontal cortex (PFC)
as other ways of terminating the dopamine signal, such as the presence
of the dopamine transporter (DAT), are limited in this region (Käenmäki
et al., 2010).
Given the fundamental role of COMT in modulating prefrontal
cortical dopaminergic neurotransmission, the contribution of the
COMT gene in relation to behavior has attracted attention (Huotari
et al., 2002; Matsumoto et al., 2003; Shehzad et al., 2012; Tunbridge
et al., 2004). COMT is encoded by a single gene localized on chromo-
some 22q11.1–q11.2. A common single-nucleotide polymorphism
(SNP) Val158Met in exon 4 of the COMT gene was studied intensively
in relation to psychiatric disorders as well as in suicidal behavior and
related traits, such as aggression (Crisafulli et al., 2010; Currier and
Mann, 2008). This SNP (rs4680) affects the functional ability of the
enzyme to catabolize synaptic dopamine (Tunbridge, 2010; Witte and
Flöel, 2012), such that in human postmortem PFC tissue the Val/Val
genotype is associated with about 38% greater enzyme activity than
that evident in Met/Met homozyotes (Chen et al., 2004; Tunbridge
et al., 2004, 2006).
Several studies demonstrated associations between the COMT
Val158Met gene polymorphism and psychiatric disorders, including
major depression disorder and suicidal behavior as well as antidepres-
sant treatment responses (Baune et al., 2007; Benedetti et al., 2009;
Massat et al., 2004; Ono et al., 2004; Pivac et al., 2011), although contra-
dictory results have also been reported (Hosák, 2007; Schosser et al.,
2012). A meta-analysis of six related studies suggested an association
between COMT Val158Met polymorphism and suicidal behavior, and
this relationship was moderated by gender and the lethality of suicide
attempt (Kia-Keating et al., 2007). In contrast, more recent meta-
analyses that included new data questioned the association between
COMT Val158Met and suicidal behavior (Calati et al., 2011; Tovilla-
Zarate et al., 2011). Although more than 16 case–control studies have
been reported, the association between COMT Val158Met polymor-
phism and suicidal behavior remains controversial, possibly owing to
different end points (suicide vs. suicidal intent) that have been used in
these analyses.
Since suicidal behavior ranges from suicidal ideation to suicide
attempts and completed suicide, it is possible that depressed individuals
that completed suicide, obviously the most severe form of suicidal
behavior, represent a more homogenous group than groups that include
suicidal ideation and suicide attempts (suicide attempts with non-fatal
outcomes) (Kia-Keating et al., 2007; Supriyanto et al., 2011). Indeed, as-
sociation studies suggested that the lethality of suicide attempt may be
associated with certain gene polymorphism, such as 5-HTTLPR (Gonda
et al., 2011) and COMT Val158Met (Kia-Keating et al., 2007). However,
only two studies concerning the association between COMT Val158Met
polymorphism and completed suicide have been reported (Ono et al.,
2004; Pivac et al., 2011). Given the different outcomes observed in the
populations studied, additional analyses of completed suicides are
needed in order to determine whether there is an association between
COMT Val158Met polymorphism and suicidal behavior, and important-
ly, to determine whether the Val158Met polymorphism is, in fact, relat-
ed to altered COMT mRNA expression levels. In this regard, there have
been few reports documenting the relationship between the Val158Met
Table 1
Age and cause of death among male and female control and suicide participants in the association study.
Subjects Age (year)
Control female 67.35 ± 17.23
Control male 61.65 ± 14.60
Suicide female 49.77 ± 16.89
Suicide male 48.06 ± 15.28
ACF = acute cardiac failure; AMI = acute myocardial infarction; TC = traffic accident; poisoning (includes prescription drug overdose).
179
polymorphism and COMT mRNA expression levels. COMT mRNA ex-
pression in human brain was found to be related to a haplotype contain-
ing Val158Met in schizophrenia, bipolar disorder and major depression
(Bray et al., 2003; Dempster et al., 2006; Zhu et al., 2004), but, the ab-
sence of a correlation between COMT genotypes and expression of its
mRNA was also reported (Chen et al., 2004; Tunbridge et al., 2004). In-
terestingly, Matsumoto et al. (2003) found a disease-related laminar
difference of COMT expression in the dorsolateral prefrontal cortex
(DLPFC) of patients with schizophrenia; however, the relation between
the COMT Val158Met polymorphism and COMT expression levels with-
in the depressed suicide brain is still uncertain.
The goal of the present study was to determine whether or not
differences existed in the distribution of COMT Val158Met polymor-
phism in depressed individuals that died by suicide relative to that evi-
dent in nondepressed individuals that died of causes other than suicide.
As well, in a subset of samples from the FPC and OFC we assessed poten-
tial differences of COMT mRNA expression in completed suicide relative
to control subjects, and determined the relation between COMT
Val158Met polymorphism and COMT mRNA expression in both of
these conditions.
2. Materials and methods
All procedures, including tissue harvesting, were approved by the
local ethics committee (Semmelweis) and the Research Ethics Board
of the Royal Ottawa Health Care Group approved analyses of tissue
samples at University of Ottawa Institute of Mental Health Research.
Analyses were performed in accordance with the Declaration of
Helsinki. Tissue harvesting occurred after written informed consent
was obtained from next of kin, which included the request to consult
medical charts and to conduct neurochemical and/or biochemical
analyses.
2.1. Subjects
Brains from 49 depressed individuals that died by suicide (n = 35
males and 14 females) and from 72 control participants (n = 46
males and 26 females), who died from causes not directly involving
any diseases of the central nervous system, were obtained at autopsy
at the Department of Forensic Medicine of the Semmelweis University
Medical School (Budapest), at the Department of Neuropathology,
National Institute of Psychiatry and Neurology, Budapest, and at the
Department of Pathology of Saint George Hospital, Székesfehérvár,
Hungary. The microdissected brains were stored in the Human Brain
Tissue Bank, Budapest. Tissue samples from some of these brains were
previously used in studies assessing serotonin receptors (Anisman
et al., 2008), and epigenetic differences regarding the GABAA subunits
(Poulter et al., 2008, 2010).
Table 1 provides the age and cause of death of the control partici-
pants and those that died by suicide for which genotyping was conduct-
ed. Of these participants, only a subset was used for the determinations
of COMT mRNA expression in the FPC and OFC. The sample comprised 3
independent sets of tissue obtained over a 4 year period. Table 2 shows
the sex, age, postmortem interval and cause of death for each sample
that went into the qPCR analysis. Overall, control participants used
for genotyping were older than those that died by suicide [F (1,
Cause of death
n = 26: 17 ACF, 3 AMI, 2 stroke, 4 bronchopneumonia
n = 46: 31 ACF, 10 AMI, 2 TC, 1 stroke, 1 electric shock, 1 CO intoxication
n = 14: 6 hanging, 8 poisoning
n = 35: 24 hanging, 3 jumping, 8 poisoning
180 L. Du et al. / Progress in Neuro-Psychopharmacology & Biological Psychiatry 50 (2014) 178–183
Table 2
Demographic information of control and suicide participants in COMT gene expression analysis.
Region Subjects n Gender
FPC Controls 27 15F/12M
Suicides 29 11F/18M
OFC Controls 15 4F/11M
Suicides 19 9F/10M
ACF = acute cardiac failure; AMI = acute myocardial infarction; other causes included bronchopneumonia, electric shock, asphyxiation (smoking in bed), traffic accident, Guillain–Barre
syndrome, brain hemorrhage and myodegeneration; poisoning (includes prescription drug overdose).
120) = 24.23, p b 0.01], as they were with respect to samples from FPC
[F (1, 54) = 3.63, p = 0.06], and for the OFC [F (1, 32) = 4.13,
p = 0.05]. However, the age differences between controls and suicide
subjects had no influence on COMT Met168Val genotypic distribution
and the overall statistical results.
Suicides and controls were Caucasian from the Budapest region
(Hungary). Medical, psychiatric and drug histories of those who died
by suicide were obtained through interviews with the attending
physician/psychiatrist and family members, and through chart reviews.
The postmortem diagnoses were conducted and/or confirmed by
experienced psychiatrists on the basis of DSM-IV criteria (American
Psychiatric Association, 1994). For all individuals that had died by
suicide a psychiatric diagnosis of depressive disorder was on record.
Based on these interviews and chart reviews, participants were only
included if they had not used antidepressant medication for at least
two months prior to death and did not have a history of either drug
or alcohol abuse. The absence of antidepressants was confirmed by
toxicological tests of blood samples. As well, toxicological analyses of
blood samples confirmed that drugs or alcohol was not present. In
49 cases, a psychiatric diagnosis of affective disorder was on record.
These included diagnoses of recurrent major depression-unipolar
(n = 47), bipolar illness-depressed phase (n = 1) and depression with
psychotic features (n = 1) that had been established by experienced
psychiatrist on the basis of DSM-IV criteria.
Interviews with family members, together with examination of
medical records, indicated that control participants had never been
treated for depression, and confirmed the absence of a history of psychi-
atric illness, as well as alcohol or drug abuse during the last ten years.
Subjects with a history of schizophrenia, epilepsy, alcohol abuse or
other drug abuse were excluded from this study.
2.2. Tissue collection, dissection and storage
Brain tissue was obtained 1–16 h after death. Tissue from suicides was
obtained later than controls [M ± SEM = 4.26 ± 0.25 and 3.44 ±
0.37 h, respectively; F (1, 54) = 3.33, p = 0.07] for FPC and OFC
[M ± SEM = 5.71 ± 0.65 vs. 4.75 ± 0.43; F (1, 32) = 1.54, p = 0.22],
but within this narrow window mRNA expression for COMT was not
significantly correlated with the time to tissue harvesting and inclusion
or exclusion of PMI as a covariate had no effect on the findings. Brains
were removed from the skull and cut into six major sections (four cortical
lobes, basal ganglia-diencephalon and lower brain stem-cerebellum),
rapidly frozen on dry ice, and stored at −70 °C until dissection (2 days
to 2 months later). At dissection the brain samples were sliced into
1–1.5 mm thick coronal sections at a temperature of 0–10 °C. Using a
fine microdissecting (Graefe's) knife the right frontopolar cortex
(Brodmann area 10) was dissected at the most polar portion of the
frontal lobe below the intermediate frontal sulcus, and the orbital
frontal cortex. The orbital frontal cortex included the anterior orbital
gyrus and the anterior parts of the medial and lateral orbital gyri.
These corresponded to Brodmann area 11 and the ventral part of
Brodmann area 12. The tissue samples did not contain any parts of the
gyrus rectus, the posterior orbital gyrus or the neighboring Brodmann
areas 45 and 47. Tissue samples were stored in airtight containers or
Age (year) Cause of death PMI (h)
60.63 ± 14.19 17 ACF, 4 AMI, 2 stroke, 1 pneumonia, 3 other 3.44 ± 1.95
53.59 ± 13.48 16 hanging, 11 poisoning, 2 jumping 4.26 ± 1.35
61.27 ± 15.63 7 ACF, 4 AMI, 4 other 4.70 ± 1.56
50.74 ± 14.49 11 hanging, 6 poisoning, 2 jumping 5.71 ± 2.83
plastic tubes at −70 °C until further use. Cortical samples were always
taken from the right hemisphere.
2.3. DNA extraction and COMT Val158Met genotyping
DNA was extracted from postmortem brain tissue using standard
salting-out procedures as described in detail previously (Du et al.,
1999). COMT Val158Met polymorphism (rs4680) was genotyped by po-
lymerase chain reaction (PCR) followed by restriction enzyme digestion
as described by Daniels et al. (1996) with minor modifications. A 169-bp
segment was amplified by PCR on a thermocycler. The following primers
were used: sense 5’-ACTGTGGCTACTCAGCTGTG-‘3 and antisense 5’-
CCTTTTTCCAGGTCTGACAA-3’. PCR products were digested by restriction
enzyme NlaIII (New England Biolabs, Whitby, Ontario) and then subject-
ed to electrophoresis in a 10% polyacrylamide gel. Digestion products
were visualized by ethidium bromide staining under ultraviolet light.
2.4. RNA extraction and reverse transcription-real-time quantitative PCR
(RT-qPCR)
Samples that had been stored in air-tight containers at −70 °C were
thawed slowly, after which total brain RNA was isolated and purified
by standard methodologies employing Trizol according to the
manufacturer's protocol (Invitrogen, Burlington, Ontario). Isolated RNA
was checked for purity by ensuring that the OD 260/280 ratio was great-
er than 1.8. Analyses of the RNA quality using Agilent BioAnalyzer
showed little degradation. Samples for qPCR analyses were prepared
by reverse transcribing 5.0 μg of total RNA using Superscript II reverse
transcriptase (Invitrogen; Burlington, Ontario). Aliquots of this reaction
were then used in simultaneous qPCR reactions. The RNA integrity num-
ber (RIN) for controls was 5.96 ± 0.32 (range = 5.2–8.5), whereas in
suicide samples the RIN was 6.57 ± 0.55 (range = 5.2–8.9) (F b 1.0).
The correlation of the RIN versus the cycle threshold of glyceraldehyde
3-phosphate dehydrogenase (GAPDH) was uniformly low in the control
and suicide samples. The tissue pH for controls and depressed/suicide
samples were 6.67 ± 0.26 (range = 5.9–7.1) and 6.57 ± 0.28
(range = 6.1–7.14), respectively (p N 0.1).
For qPCR, SYBR green detection was used according to the
manufacturer's protocol (iTaq™ SYBR® Green Supermix with ROX,
Bio-Red, Mississauga, Ontario). A Stratagene MX-4000 real time
thermocycler was used to collect the data. The COMT PCR primer pairs
(sense primer: 5’-ACCTACTGCGAGCAGAAG-3’ and antisense primer:
5’-CACAGCTGAGTAGCCACAG-3’) generated a 162 base pairs amplicon.
Primer efficiency was measured, using MX-4000 software, from the
slope relation between a dilution series of reverse-transcribed RNA
(cDNA) and the cycle threshold (Ct value). The COMT primer pairs
had an efficiency over 95%. For the qPCR analysis, primers that amplify
GAPDH (sense primer: 5’-AGCCTCAAGATCATCAGCAATG-3’ and anti-
sense primer: 5’-GTCTTCTGGGTGGCAGTGATG-3’) mRNA were used as
housekeeping genes to normalize the data. The COMT and GAPDH
primers have similar amplification efficacies. The Ct of COMT gene
product was normalized against that of the reference gene GAPDH,
which was run simultaneously. The ΔCt for each sample was calculated
as ΔCt = [Ct (COMT) − Ct (GAPDH)]. To quantify mRNA expression
levels, the 2-ΔΔCT method (Livak and Schmittgen, 2001) was used to
 L. Du et al. / Progress in Neuro-Psychopharmacology & Biological Psychiatry 50 (2014) 178–183
convert ΔCt values to mRNA fold changes relative to the female control
group (calibrator).
2.5. Statistical analyses
All statistical analyses were performed using the Statistical Package
for Social Sciences (SPSS) for windows version 11.5 (SPSS, Chicago,
IL). Allele and genotype frequencies were compared between depressed
suicides and control subjects by a chi-square analysis. Hardy–Weinberg
equilibrium for the distribution of genotypes was tested with the chi-
square test for quality of fit. The level of significance was set at p = 0.05.
Effects of Val158Met variation on gene expression were examined
using ANCOVA with genotype and diagnosis as independent variables
and age and postmortem interval as covariates. The mRNA expression
of COMT was analyzed by a 2 (Suicide vs. Control) × 2 (Male vs.
Female) ANOVA. Using Grubb's test, one identified outlier was deleted.
Deleting this one outlier did not change overall statistical results.
3. Results
3.1. Association study
Genotypic distributions of those that died by suicide and control
subjects were in Hardy–Weinberg equilibrium (χ2 = 2.44, p = 0.12,
and χ2 = 2.36, p = 0.13, respectively). No significant differences
were observed between suicides and controls in genotypic distribution
(χ2 = 0.06, p = 0.97) and allelic frequencies (χ2 = 0.01, p = 0.92).
Likewise, the genotypic distribution did not differ when the analysis
included gender (Table 3).
3.2. COMT expression in the frontopolar cortex and orbital frontal cortex
The age of participants at the time of death and postmortem interval
(PMI) were not significantly correlated with COMT mRNA expression
(r = − 0.10, p = 0.46 and r = 0.13, p = 0.33, respectively) in the
FPC. Analyses of COMT mRNA expression within the FPC indicated
that expression levels were significantly higher in suicides than in con-
trols [F (1, 54) = 15.29, p = 0.001]. There was no expression difference
between genders [F (1, 52) = 0.02, p = 0.88], and no Group × Gender
interaction was detected [F (1, 52) = 0.09, p = 0.76]. Altered COMT
mRNA expression was also observed in the OFC [F (1, 32) = 13.17,
p = 0.001]. The levels of COMT mRNA expression in this region were
significantly increased in individuals that died by suicide relative to
those that died of other causes, being up-regulated by approximately
60% and 65% in the FPC and OFC, respectively.
3.3. Relationship between Val158Met genotype and COMT expression
Since prior studies on the relation of suicide and COMT gene focused
on gene polymorphisms rather than on gene function, we also explored
Table 3
Genotype and allele distributions of COMT 158Val/Met polymorphism in depressed/suicide individuals and nondepressed individuals.
COMT Val108/158Met
Genotypes
Val/Val (%) Val/Met (%)
Controls 22 (30.5) 30 (41.7)
Suicides 16 (32.7) 19 (38.8)
χ2 = 0.11; df = 2; p = 0.95
Male controls 15 (31.9) 18 (38.3)
Male suicides 13 (37.1) 16 (45.7)
χ2 = 1.74; df = 2; p = 0.42
Female controls 7 (29.2) 11 (45.8)
Female suicides 3 (21.4) 3 (21.4)
χ2 = 4.13; df = 2; p = 0.13
181
the relation between COMT Val158Met genotypes and their mRNA
expression in the FPC and OFC. No association was found between
genotype and mRNA expression [F(2,53) = 0.55, p = 0.58], and gender
had no influence in relation to COMT expression [F(1,54) = 0.24,
p = 0.63] in the analyses of either the FPC or the OFC in the pooled
suicides and controls [mRNA expression: F(2,31) = 0.77, p = 0.47
and gender effect: F(1,32) = 3.3, p = 0.08]. Importantly, however, a
Group (suicides or controls) × Genotype interaction on COMT expression
was apparent within the FPC [F(2,50) = 5.46, p = 0.007; Table 4].
Individuals with the Met allele (Met/Met and Met/Val genotypes) who
died by suicide displayed higher COMT mRNA expression than did the
Met allele carriers who were not depressed (Fig. 1). A similar difference
was not apparent among those carrying the Val/Val genotype. In contrast
to the FPC, there was no Group × Genotype interaction on gene expres-
sion in the OFC [F(2,28) = 2.1, p = 0.14]. Correlational analyses also
revealed that genotype was correlated with COMT gene expression in
the FPC of control samples (Spearman's rho = 0.63, p = .0001), but
not in the FPC of suicide samples (Spearman's rho = −0.25, p = 0.19).
In the OFC, significant correlations were not detected in either controls
or suicides (p N 0.30; data not shown).
4. Discussion
As reported in recent meta-analyses relating COMT polymorphism
and suicidal intentions (Calati et al., 2011; Tovilla-Zarate et al., 2011),
in the present study, significant associations were not observed
between suicide completers and either the genotype distribution or
allelic frequencies of the COMT Val158Met polymorphism. To our
knowledge, only two prior studies investigated the association between
COMT Val158Met and suicide completion, as opposed to suicide
attempts (Ono et al., 2004; Pivac et al., 2011). It had been suggested
based on a Japanese sample that the Val/Val genotype served as a
protective factor against suicide in males, as the Val/Val genotype
occurred at a higher frequency in male controls than in male suicides.
In contrast, Pivac et al. (2011) showed that in a Caucasian population
of Slovenian origin there was an overrepresentation of the Met/Met in
male control subjects compared with males that died by suicide,
suggesting that Met/Met genotype might be a protective factor against
suicide. The discrepancies between these studies and with that of the
present investigation in which an association was not observed
between the genotype and the occurrence of suicide might be due to
any number of factors. Significantly, the suicide cases in the present
study were determined on the basis of psychological autopsy and
included only individuals who had been depressed, whereas the two
earlier studies also involved individuals that had died by suicide, but
without a psychological autopsy having been performed for diagnostic
purposes. Thus, it is uncertain whether these studies, in fact, involved
suicides related to depression or suicides associated with other condi-
tions. Furthermore, ethnic differences in genotype and allele frequency
distributions exist between Caucasian and Japanese male control
Alleles
Met/Met (%) Val (%) Met (%)
20 (27.8) 74 (51.4) 70 (48.6)
14 (28.6) 51 (52.0) 47 (48.0)
χ2 = 0.01; df = 1; p = 0.92
14 (29.8) 48 (51.0) 46 (49.0)
6 (17.1) 42 (60.0) 28 (40.0)
χ2 = 1.29; df = 1; p = 0.25
6 (25.0) 25 (52.1) 23 (47.9)
8 (57.1) 9 (32.1) 19 (67.9)
χ2 = 2.84; df = 1; p = 0.09
182 L. Du et al. / Progress in Neuro-Psychopharmacology & Biological Psychiatry 50 (2014) 178–183

Table 4 greater stress was associated with lower methylation of the Val158 al-
ANCOVA analysis of COMT gene expression in the PFC. lele in peripheral blood mononuclear cells of Val/Val individuals
Brain region Variable F df p (Ursini et al., 2011).
In the current study, we detected an alteration of COMT mRNA
FPC (n = 56) Diagnosis (suicides vs. controls) 15.29 1 b0.001⁎⁎⁎
Val158Met 0.55 2 0.58 expression in the PFC of suicide victims, but significant associations
Gender 0.24 1 0.63 were not observed in the COMT Val158Met genotypic distribution or
Diagnosis × Val158Met 5.45 2 0.007⁎⁎ allelic frequencies between suicide completers and controls. The lack
Diagnosis × Gender 0.09 1 0.76
of an association may be due to small sample size. It is also possible
OFC (n = 34) Diagnosis (suicides vs. controls) 13.77 1 0.001⁎⁎
Val158Met 1.11 2 0.34 that no associations may indicate that the Val158Met polymorphism is
Gender 3.29 1 0.08 most likely not a major player in the genetic susceptibility to suicide.
Diagnosis × Val158Met 2.11 2 1.40 Our results agree with previous reports that the levels of COMT mRNA
Diagnosis × Gender 0.11 1 0.74 expression were elevated in schizophrenia, bipolar disorder and suicide
⁎⁎ p b .01. (Abdolmaleky et al., 2006; Matsumoto et al., 2003), suggesting that
⁎⁎⁎ p b 0.001. increased dopamine degradation in the PFC plays a role in relation
with suicide.
samples (Pivac et al., 2011), which might also have contributed to the The sample size in the present study was admittedly small, but the
differences observed across studies. frequencies of the COMT Val158Met genotype were similar to that of a
Although the COMT Val158Met variant has been shown to have previous report in healthy East European controls (Nedic et al., 2010).
major effects on enzyme activity, its actions on COMT mRNA expression This said, suicide is a complex behavior that likely involves multiple
levels in the FPC and OFC of depressed suicides have not previously been genes and gene × environment interactions, and hence it can be
examined. It has been shown that a haplotype that contained the Val expected that any single variable would have small to modest genetic
allele was associated with reduced COMT expression in normal effects in relation to completed suicide (Kendler, 2010). Although the
human brain (Bray et al., 2003), and it was also demonstrated that the present study, like many association studies, has limited power to detect
Val158Met genotype was positively related to COMT gene expression small effects, it may nevertheless contribute to future (meta-) analyses
in the cerebellum (Dempster et al., 2006; Zhu et al., 2004). However, that combine several studies in an effort to overcome difficulties related
this polymorphism in the dorsolateral prefrontal cortex was unrelated to small sample sizes.
to major depression, schizophrenia, and bipolar disorder, (Chen et al., In addition, the alteration of COMT mRNA levels in suicide brains
2004; Tunbridge et al., 2004). In the present investigation the COMT might also be caused by altered mRNA expression and/or degra-
Val158Met variant was related to COMT mRNA expression levels in dation rates owing to other factors, such as methylation levels and
the controls, but not in suicide brains. Indeed, among those that died transcriptional/post-transcriptional regulation, as well as that mRNA
by suicide who carried the homozygous Met/Met alleles, higher COMT expression do not necessarily reflect the final protein levels/expression.
mRNA expression was evident than in those who carried Val/Met and Summarizing, the present study revealed that COMT mRNA expres-
Val/Val genotypes, whereas in controls Met/Met carriers expressed sion levels were significantly higher in the FPC and OFC of depressed
lower COMT mRNA. This is in agreement with previous findings individuals that died by suicide than among nondepressed individuals
(Abdolmaleky et al., 2006) indicating that in suicide samples (15 bipolar that died as a result of factors other than suicide. The increased COMT
and 7 schizophrenia suicides) expression of membrane-bound expression might be associated with lower dopaminergic activity in
catechol-O-methyltransferase (MB-COMT is predominantly expressed these regions, which could potentially have been related to depres-
in the brain; Tenhunen et al., 1994) was higher than that in controls, sion/suicide. Several studies have indeed suggested an association
possibly owing to hypomethylation of the MB-COMT promoter. In this between dopaminergic hypoactivity and suicide. For example, decreased
particular study the Met homozygosity was accompanied by approxi- dopaminergic neurotransmission has been reported in depressive
mately two-fold more COMT mRNA in those that died by suicide relative disorders (see reviews by Dailly et al., 2004; Dunlop and Nemeroff,
to controls. It is uncertain whether the higher COMT mRNA expression 2007; Pitchot et al., 2001) and reduced dopamine turnover was
in suicide samples of the present study was due to hypomethylation observed in the basal ganglia of depressed suicides (Bowden et al.,
of the COMT promoter. However, given that depression and suicide 1997) and CSF of depressed suicide attempters (Sher et al., 2006). To
are associated with considerable distress, it might be significant that be sure, being correlational, the present data, like those of earlier
studies, do not speak to whether dopamine plays a causal role in
suicide/depression. Moreover, it remains to be established whether
the altered COMT mRNA expression in the present study was related
to depression or was unique to individuals that died by suicide. Never-
theless, the data available are in line with the view that dopamine
functioning plays a role in relation to depression/suicide (Berton et al.,
2006; Nestler and Carlezon, 2006).

Acknowledgments

This research was supported by an Ontario Mental Health Foundation

operating grant to LD, HA, and ZM and by the European Union Grant
No. FP6, BNEII No. LSHM-CT-2004-503039 to MP. HA holds a Canada
Research Chair. We thank Yue He and Jill Chatt for their technical assis-
tance. The authors report no biomedical financial interests or potential
conflicts of interest.

Appendix A. Supplementary data

Fig. 1. Normalized COMT mRNA expression in FPC of depressed suicide victims and control
subjects with genotypes Met/Met (control: n = 10 vs. suicide: n = 6), Met/Val (control:
n = 11 vs. suicide: n = 12), and Val/Val (control: n = 6 vs. suicide: n = 11). *p b 0.05; Supplementary data to this article can be found online at http://dx.
**p b 0.01. doi.org/10.1016/j.pnpbp.2013.12.016.
 L. Du et al. / Progress in Neuro-Psychopharmacology & Biological Psychiatry 50 (2014) 178–183
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