Number of risk genotypes is a risk factor for major depressive disorder: a case control study
© Garriock et al. 2006
Received: 10 April 2006
Accepted: 05 July 2006
Published: 05 July 2006
The objective of the study was to determine the genetic basis of Major Depressive Disorder, and the capacity to respond to antidepressant treatment. An association study of 21 candidate polymorphisms relevant to monoamine function and the mechanism of antidepressant response was conducted in 3 phenotypically distinct samples: a group with chronic or recurrent depression unable to respond to antidepressants (non-responders) (n = 58), a group capable of symptomatic improvement with or without treatment (responders) (n = 39), and volunteer controls (n = 85). The responders and non-responders constituted a larger group of depressed subjects.
A candidate gene approach was employed to asses the genetics basis of Major Depressive Disorder. The genotypic frequencies of selected polymorphisms were compared between the controls and depressed subjects. To asses the genetics basis of the capacity to respond to antidepressant treatment, the responders were compared to the non-responders. Candidate genes were chosen based on functional studies and proximity to whole genome linkage findings in the literature. Risk genotypes were identified by previous functional studies and association studies.
A statistically significant difference in genotype frequency for the SLC6A4 intron 2 VNTR was detected between the subjects with a history of depression and controls (p = 0.004). Surprisingly, a statistically significant difference was detected between responders and non-responders for the DRD4 exon III VNTR genotype frequencies (p = 0.009). Furthermore, a difference between the controls and depressed subjects as well as between the controls and non-responders was detected for the number and distribution of risk genotypes in each group.
An association between several monoamine-related genes and Major Depressive Disorder is supported. The data suggest that the two depressive phenotypes are genetically different, inferring that the genetic basis for the capacity to respond to standard antidepressant treatment, and the genetic susceptibility to Major Depressive Disorder may be independent. In addition, a proof of concept is provided demonstrating that the number of risk genotypes may be an indication of susceptibility of major depressive disorder and the severity of the disorder.
variable number of tandem repeats
single nucleotide polymorphism
treatment resistant depression
dopamine receptor D4
Antidepressant Treatment History Form
polymerase chain reaction
diagnostic and statistical manual
Structured Questionnaire Interview for DSM-IV-R
Depression affects more than 40 million Americans at some time during their lives and represents one of the most debilitating medical conditions. A substantial body of evidence drawn from a range of methods, proband groups, and diagnostic criteria has established that a familial phenotype is present in patients with major depression [1–7]. Despite their limitations, molecular methods such as candidate gene association studies also support the concept of a genetic influence in vulnerability to depression [8, 9]. Although genetic studies promise to improve our understanding of the pathophysiological and genetic aspects of the depressive syndrome, such understanding is limited by the genetic complexity and lack of discrete, etio-pathologically related, and homogeneous "depression" phenotypes.
Treatment resistant depression (TRD) is a characteristic of some depression phenotypes. It can range from not being able to respond to one medication, to not clinically responding to electroconvulsive therapy. TRD, or non-response can be viewed as a spectrum of severity of major depressive disorder, as the amount of suffering is greater for those with TRD, than those without. Whether or not TRD is a subtype of depression with a distinct underlying biology is still unknown and under investigation. It is possible that to some extent TRD is a result of hyperfunctioning enzymes that metabolize typical antidepressant medications . In this case, the TRD subjects would have a genetic background that leads to a pharmacokinetic difference from the depressive group which is capable of antidepressant treatment response. This genetic discrepancy between two diagnostically similar groups (though phenotypically difference in terms of response profiles) may be detectable, and would provide insight to the biology of treatment resistant depression.
To explore genetic differences within depression endophenotypes, we genotyped Caucasian subjects with unipolar major depression from two distinct treatment response profiles: those able to respond to treatment, and those not able to respond. The genes being studied encode for receptors, transporters, synthetic or degradation enzymes, transcription factors, and neurotrophic factors related to monoamine function. We hypothesized that the depressive group as a whole would be genetically distinct from the healthy controls. It was predicted that there would be a higher frequency of "risk" genotypes (excessive or deficient in functioning depending on the case) in the depressive groups when compared to the controls.
We report one two single gene findings and one polygenic discovery. A difference in genotypic frequency between the controls and depressed subjects was detected in the serotonin transporter intron 2 VNTR polymorphism. A surprising difference was detected between the responder and non-responders in the dopamine D4 receptor exon III VNTR. Furthermore, we demonstrate that depressed subjects have more risk genotypes that non-depressed subjects, and that non-responders have the most risk genotypes of all our phenotypic groups analyzed.
Clinical description and demographics of subjects groups
Descriptive clinical information
Mean years of age (range)
Combination of Non-responders and responders (see below)
Non-responders (treatment resistant depression)
Clinical diagnosis of chronic or recurrent depression and failure to achieve remission after at least two adequate antidepressant trials as documented by the Antidepressant Treatment History Form (ATHF).
Clinical diagnosis of major depressive disorder, and capability to respond to standard antidepressant medication.
Subjects who denied a personal history of mental illness based on the mood disorders section of the Structured Questionnaire Interview for DSM-IV-R (SCID) questionnaire.
Type of Polymorphism
location in gene; details
intron 2 near 3' splice site of exon 2; SLC6A4, HTR2A, DbH, DRD1, SLC6A3 have Ap2B binding sites
G-->A SNP at nucleotide 196; rs6265;
proBDNF coding region; (val66met)
44 bp in/del (5-HTTLPR)
Promoter; actually a VNTR (xs-xl)
VNTR of 17 bp element (9–12 copies)
Dopamine receptor D4
120 bp in/del
0 or 1 7R
Dopamine receptor D4
48 bp VNTR
Serotonin post-synaptic receptor 2A
C102T SNP; rs6313
9/9 and 9/10
Dopamine transporter (SLC6A3)
40 bp VNTR (9 and 10 repeats most frequent, but 3–11 copies possible)
3' non-coding region of exon 15
G-->A SNP (MspI); rs165599
3'-UTR; maybe in mRNA
Dopamine receptor D3
G-->A SNP (Glycine--> serine) (MscI, isoschizomer of BalI) rs6280
exon 1 (N-terminal extracellular domain)
Dopamine Recptor D1
G-->C SNP (HaeIII)
Dopamine Recptor D1
T-->C SNP (Bsp1286I)
Dopamine Recptor D1
T-->C SNP (HaeIII)
Serotonin receptor 6A
T267C SNP (RsaI) Tyr(89) silent mutation; rs1805054
coding region (1st extracellular loop)
Dopamine Recptor D1
G-->A SNP (DdeI)
G-->A SNP (va158met) (NlaIII) rs4680
4th exon (codon 158 or 108)
Monoamine Oxidase type A
30 bp VNTR (2R, 3R, 3.6R, 4R, 5R, 7R)
1.2 kb upstream of coding region; promoter
Serotonin Receptor 1A
C-->G SNP (BstF5I) at position -1019 from ATG start site
PCR fragment is from -1158 to -996 from ATG start site strobel, 2003; promoter
C-->T SNp at position 1604; R535C (BstUI); rs6271
exon 11: 142 bp fragment (originally FnuDII digest). A1: 95, 47 bp; A2: 66, 47, 29 bp.
Tryptophan Hydroxylase 2 (neuronal TPH)
Initially, four comparisons were made: between subjects with a history of major depression vs. controls, non-responders vs. controls, responders vs. controls, and non-responders vs. responders. The data presented in this study is a result of a chi-square analysis was used to test for differences in the frequency of genotype polymorphisms between the depressive subjects and healthy controls as well as between the responders and non-responders. The remaining two comparisons (responders vs. controls, non-responders vs. controls) is available upon request. All significance tests (aside from QVALUE) were done with SPSS version 14. Genotypic frequency differences were analyzed by Chi-Square tests. False discovery rate analysis was used to control for multiple comparisons between groups and number of polymorphisms. The q-values are reported with all p values ≤0.05. A q-value is an indication of the percent of time a Type I error would be committed if a corresponding p-value was considered as statistically significant. For example, we accepted a false discovery rate of 15%, thus based on the value distribution of the present dataset, a value of q ≤0.137 was considered statistically significant, however, this still implies that 13.7% of the time when we accept a corresponding p-value as statistically significant, an error would be made. The statistical testing for each polymorphism was between a risk genotype determined a priori to testing (see Table 2), and the remaining genotypic groups. The mean number of risk genotypes among the phenotypic groups was analyzed using independent samples t-tests. We chose this approach because we had hypotheses that the history positive group would have more risk genotypes that the history negative, and likewise, that the non-responders would have more risk genotypes that the non-responders.
P and Q-values for trending and statistically significant (after multiple comparison correction *) polymorphisms
Depressed vs. non-depressed
Responders vs. non-responders
STin2 VNTR SLC6A4
exon 3 VNTR DRD4
The genetic susceptibility to major depression was tested using genotype frequency comparisons between subjects with a history of depression and controls. We detected three polymorphisms (SLC6A4 intron 2 VNTR, 2 COMT polymorphisms) which may contribute to the genetic susceptibility to major depression, however, only one (SLC6A4) which held up to multiple comparison corrections.
The comparison between the non-responders and responders is a method to detect the genetic basis for antidepressant response, in general. Out of the 21 polymorphisms tested, only one (DRD4 exon 3 VNTR) was statistically significant. This is compared to the 3–4 polymorphisms which demonstrate a genetic susceptibility to depression. It was unpredicted that the genetic basis for antidepressant response may differ from the genetic basis of susceptibility to major depression.
We further demonstrated that the number of risk genotypes is not consistent across phenotypes. The non-responders have the most risk genotypes, followed by (in decreasing order) the depressed group as a whole, the responders, and the controls. This supports the notion that Major Depressive Disorder is polygenic, and that the number of risk genotypes may be an indication of the susceptibility to Major Depressive Disorder as well as the severity of the disorder.
In retrospect, we recognize that much of our current understanding of the pathophysiology of depressive disorders has been inferred from the prevailing hypotheses of mechanisms for antidepressant action. Accordingly, candidate genes selected for their function are commonly associated with monoamine function or their putative intracellular responses to neurotransmitter activation. The neurobiology of treatment resistance is not well understood; it may represent an extreme phenotype along a unique pathological continuum of depression. This interpretation may be supported by the fact that the usual monoamine-based treatment interventions, by definition, provide little or no benefit to patients with treatment resistant depression.
The Hamilton Depression Rating Scale has been for decades the gold standard of depression symptoms quantification and has been utilized in >95% of antidepressant treatment studies. There have been lately a number of concerns primarily about the scale's ability to reliably quantify antidepressant responses given the multi-dimensionality of the scale, which may non-specifically determine response in individuals treated with sedating agents such as TCA's. In the other hand, given the number of non-melancholic specific items, it may be less sensitive at detecting an antidepressant response. It is possible that alternative tools such as the Bech Rafelsen or the Montgomery Asberg Depression Rating Scale among others may more accurately reflect antidepressant responses leading to a more reliable detection of therapeutic effects.
Genotype frequencies for polymorphisms in Table 3 for all phenotypic groups. Risk genotype indiciated by an *.
STin2 VNTR SLC6A4
Exon 3 VNTR DRD4
Despite the modest sample size, these data support the existence of a genetic basis to the susceptibility to major depression. Furthermore, depending on the phenotypic definition used in testing, different associations may be detected. Given that we utilized phenotypic definitions based on patters of treatment response, these findings further suggest that the genetic basis for the capacity to respond to monoamine-based antidepressants is different from that of susceptibility to major depression. These data provide a proof of concept that major depressive disorder is a polygenic disorder, and that the number of risk genotypes may be an indication of susceptibility to the disorder and the severity.
1. Funded in part by National Institute of Mental Health (NIMH) Grant R01-MH066235 to Dr. Moreno, and R01-MH48977 to Dr. Delgado; Cyberonics Inc., and the Arizona Hispanic Center of Excellence.
2. Treatment resistant depression subjects were a sub-group of the pilot and pivotal studies of vagus nerve stimulation (VNS) therapy for treatment-resistant depression, sponsored by Cyberonics, Inc.; Houston, Tex.
3. Treatment Responders and Healthy controls are a sub-group of subjects who participated in NIMH Tryptophan Depletion Studies mentioned above.
4. The authors thank Susan E. Siefert, ELS, CBC, of Cyberonics, Inc. for her comments on the manuscript.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.