Iwona Małecka

Influence of DRD2 and ANKK1 polymorphisms on the manifestation of withdrawal syndrome symptoms in alcohol addiction

BACKGROUND We investigated the relationship between withdrawal syndrome symptoms and dopamine receptor 2 DRD2 gene polymorphisms-141 C I/D (rs1799732) exon 8 G/A (rs6276) and ANKK1 (Ankyrin Repeat and Kinase Domain Containing 1) gene polymorphism Taq1A (rs1800497). MATERIAL A total number of 213 patients who met the ICD 10 criteria for given phenotypes were enrolled in the study. Those phenotypes included: dissocial personality disorder, early onset, alcohol withdrawal syndrome with seizures, alcohol withdrawal syndrome with delirium tremens, and alcohol withdrawal syndrome with seizures and delirium tremens. RESULTS Our results show statistically significant associations between SNP in exon 8 A/G in the DRD2 gene and alcohol withdrawal syndrome with seizures, and between SNP in promoter -141 C I/D in the DRD2 gene and early onset of alcohol dependence (AD). The A/A genotype in exon 8 A/G polymorphism seems to be a positive predictive factor for the presence or the lack of seizures in alcohol withdrawal syndrome. The A/G genotype is possibly a protective factor for this AD phenotype. CONCLUSIONS These results suggest that both investigated DRD2 polymorphisms have an impact on the AD phenotype. The findings of the presented study reconfirm that dopamine receptor 2 gene polymorphisms are associated with alcohol addiction and alcohol withdrawal syndrome.

Family-Based and Case-Control Study of Glutamate Receptor GRIK3 Ser310Ala Polymorphism in Alcohol Dependence

Aim: The aim of this study was to evaluate the role of the glutamate receptor subunit-7 (GluR7, GRIK 3) rs6691840 (Ser310Ala, T928G) in the pathogenesis of alcohol dependence (AD). Methods: DNA was provided from AD patients (n = 209) and healthy control subjects (n = 308) all of Polish descent. The history of alcoholism was obtained using the Polish version of the SSAGA (Semi-Structured Assessment for the Genetics of Alcoholism). We conducted case-control association study and transmission disequilibrium test (TDT). GRIK3 functional polymorphism was genotyped by the PCR-RFLP method. Results: Analyses revealed that polymorphism Ser310Ala of GRIK3 gene is not associated with AD or any of its subgroups. TDT reveled an adequate transmission of both alleles in the group of alcohol families. Conclusions: These findings replicate and extend our previous research results that do not support the hypothesis of the role of rs6691840 in the pathogenesis of AD.

GABA-A receptor genes do not play a role in genetics of Lesch's typology in Caucasian subjects.

Introduction Leschs typology differentiates alcoholics into different treatment response subgroups. The effects of ethanol are mediated, to an important extent, via the GABA-ergic system. Material and methods We have evaluated the linkage disequilibrium patterns and haplotype frequencies of GABRG1 and GABRA2 genes in 133 alcoholics divided according to Leschs typology and in 145 matched controls. Results Besides several relationships at a threshold of statistical significance, we found no significant differences in the haplotype distribution of these genes between alcoholics and controls. Conclusions Leschs typology may not be related with the genotype of alcoholics – at least in terms of genes with an established role in the development of dependency.

Suicidal Behavior and Haplotypes of the Dopamine Receptor Gene (DRD2) and ANKK1 Gene Polymorphisms in Patients with Alcohol Dependence – Preliminary Report

Suicide is a significant public health issue and a major cause of death throughout the world. According to WHO it accounts for almost 2% of deaths worldwide. The etiology of suicidal behavior is complex but the results of many studies suggest that genetic determinants are of significant importance. In our study,- we have analyzed selected SNPs polymorphisms in the DRD2 and ANKK1 genes in patients with alcohol dependence syndrome (169 Caucasian subjects) including a subgroup of individuals (n = 61) who have experienced at least one suicide attempt. The aim of the study was to verify if various haplotypes of selected genes, comprising Taq1A, Taq1B, and Taq1D single nucleotide polymorphisms (SNP), play any role in the development of alcohol dependence and suicidal behavior. The control group comprised 157 unrelated individuals matched for ethnicity, gender,- and age and included no individuals with mental disorders. All subjects were recruited in the North West region of Poland. The study showed that alcohol dependent subjects with a history of at least one suicidal attempt were characterized by a significantly higher frequency of the T-G-A2 haplotype when compared to individuals in whom alcohol dependence was not associated with suicidal behavior (p = 0.006). It appears that studies based on identifying correlation between SNPs is the future for research on genetic risk factors that contribute to the development of alcohol addiction and other associated disorders. To sum up, there is a necessity to perform further research to explain dependencies between the dopaminergic system, alcohol use disorders and suicidal behavior.

Molecular Analysis of the SRD5A1 and SRD5A2 Genes in Patients with Benign Prostatic Hyperplasia with Regard to Metabolic Parameters and Selected Hormone Levels

Introduction: The etiology of benign prostatic hyperplasia (BPH) has not so far been fully explicated. However, it is assumed that changes in the levels of hormones associated with aging can contribute to the development of prostatic hyperplasia. Dihydrotestosterone combines with the androgen receptor (AR) proteins of the prostate gland. Enzyme activity is based on two isoenzymes: type 1 and type 2. 5α-reductase type 1 is encoded by the SRD5A1 gene, and type 2 is encoded by the SRD5A2 gene. The aim of our study was to determine the frequency of the SRD5A1 (rs6884552, rs3797177) and SRD5A2 (rs523349, rs12470143) genes’ polymorphisms, and to assess the relationships between the genotypes of the tested mutations, and the levels of biochemical and hormonal parameters in patients with BPH. Material and Methods: The study involved 299 men with benign prostatic hyperplasia. We determined the serum levels of particular biochemical parameters—fasting plasma glucose (FPG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL) and triglycerides (TG)—by the spectrophotometric method, using ready reagent kits. The ELISA method was used to determine the levels of the following hormonal parameters and proteins: total testosterone (TT), free testosterone (FT), insulin (I), luteinizing hormone (LH), and sex hormone binding protein (SHBG). Metabolic syndrome was diagnosed. Genotyping was performed by real-time PCR. Results: We analyzed the relationships between the incidence of particular diseases and the genotypes of the SRD5A1 and SRD5A2 polymorphisms among patients with BPH. The BPH patients with the CC genotype of the SRD5A2 rs523349 and rs12470143 polymorphisms were considerably less frequently diagnosed with metabolic syndrome (MetS) (p = 0.022 and p = 0.023 respectively). Our analysis revealed that homozygotes with the CC of the SDR5A2 rs12470143 polymorphism had visibly higher HDL levels than those with the TT and CT genotypes (p = 0.001). Additionally, we found that the patients with the CC genotype of the SDR5A2 rs12470143 polymorphism had considerably higher FT levels (p = 0.001) than the heterozygotes with the CT and the homozygotes with the TT of the genetic variant analyzed in our study. Furthermore, the patients with at least one G allele of the SRD5A2 rs523349 polymorphism had significantly lower SGBG levels (p = 0.022) compared with the homozygotes with the CC genotype. The presence of at least one A allele (AA + AG genotypes) of the SRD5A1 rs3797177 polymorphism entailed notably lower serum insulin levels than those observed in homozygotes with the GG genotype (p = 0.033). Conclusions: The study described in this article shows that selected SRD5A1 and SRD5A2 polymorphisms can alter the levels of metabolic and hormonal parameters in patients with BPH. Special attention should be paid to the SDR5A2 rs12470143 polymorphism, which is associated with a change in lipid profile, as well as with the inheritance and incidence rate of MetS among these patients. An analysis of the frequency of this polymorphism among BPH patients could be useful in estimating the risk of getting ill, and planning therapies of concomitant diseases for BPH patients.

Association and family studies of DRD2 gene polymorphisms in alcohol dependence syndrome.

INTRODUCTION The human dopamine receptor 2 gene DRD2 plays a central role in susceptibility to Alcohol Dependence Syndrome (ADS). The aim of this study was to evaluate 3 single nucleotide polymorphisms: D2 (rs1076560), Tag1D (rs1800498), Tag1B (rs1079597) located in dopamine receptor 2 DRD2 gene and its role in alcohol dependence. MATERIAL AND METHODS DNA was provided from alcohol dependent (AD) patients (n=171) and healthy control subjects (n=160) all of Polish descent. The history of alcoholism was obtained using the Polish version of the SSAGA (Semi-Structured Assessment for the Genetics of Alcoholism). We conducted case-control association study and transmission disequilibrium test (TDT). Samples were genotyped using real-time PCR method. RESULTS We did not confirm the association between studied polymorphisms and alcohol dependence syndrome. TDT reveled an adequate transmission of both alleles in the group of alcohol families. CONCLUSIONS The lack of association of studied polymorphisms and ADS does not preclude its participation in the pathogenesis. Further research is needed to determine the actual contribution of DRD2 gene in the pathogenesis of alcoholism.

Associations Between the Dopamine D4 Receptor and DAT1 Dopamine Transporter Genes Polymorphisms and Personality Traits in Addicted Patients

Many factors are involved in addiction. The dopaminergic system is thought to be the key element in this process. The mesolimbic dopamine system is a crucial element in the reward system. Changes in this system are thought to be leading to substance use disorders and dependence. Therefore, for our study we chose an analysis of two polymorphisms in genes (Variable Number of Tandem Repeats in DRD4 and DAT1) responsible for dopaminergic transmission, which might be implicated in the scores of personality traits measured by the NEO-FFI test. The study group consisted of 600 male volunteers—299 addicted subjects and 301 controls. Both groups were recruited by psychiatrists; in the case group addiction was diagnosed; in the controls a mental illness was excluded. In both groups the same psychometric test and genotyping by the PCR VNTR method were performed. The results were investigated by a multivariate analysis of the main effects ANOVA. In the presented study no DRD4 main effects were found for any of the analyzed traits but the DRD4 main effects approximated to the statistical significance for the extraversion scale. However, no DAT1 main effects were found for any of the analyzed traits but the DAT1 main effects approximated to the statistical significance for the agreeability scale.These associations open new possibilities for addiction research.

The role of OPRM1 polymorphism in the etiology of alcoholism

BACKGROUND Numerous studies have investigated the association between the OPRM1 A118G polymorphism (rs1799971) and alcohol dependence, but the results have been inconsistent. The endogenous opioid system has been implicated in the development of alcohol dependence for its prominent role in the central rewarding mechanism. OBJECTIVES The aim of this study was to evaluate the role of the A118G polymorphism of the OPRM1 gene in the pathogenesis of alcohol dependence syndrome (ADS). MATERIAL AND METHODS The OPRM1 (rs1799971) polymorphism was investigated in an association study of a group of ADS patients (n = 177) and in subgroups (delirium tremens and/or seizures, age at onset <26 years, dissocial alcoholics, positive familial history of alcoholism, delirium tremens, and seizures). The control group consisted of healthy volunteers, with matched gender and age, and with psychiatric disorders excluded (n = 162). RESULTS Our research shows that there are differences in the genotypes and alleles of the OPRM1 polymorphism in the case-control study. Furthermore, we observed associations in our homogeneous subgroups - in the group of patients with ADS and accompanying delirium tremens and/or seizures at the genotype level, as well as in the subgroup of patients under 26 years of age with an early onset of dependence. CONCLUSIONS It is strongly possible that the G allele described in numerous studies can be associated with a response to treatment, but not typology, or the very predisposition toward alcoholism. It is necessary to carry out further research which would embrace a larger group of patients; it should be divided into other homogeneous subgroups, including, e.g., naltrexone pharmacotherapy.

EPA-1077 – Haplotype analysis of GABRA2 gene polymorphisms in alcohol dependence

Introduction Studies aimed at the identification of potential risk factors of the alcohol dependence syndrome (ADS) and, consequently, of risk groups of this condition that should be subjected to preventive programs, are being undertaken since many years. The aims of this study were (1) to analyze the effects of polymorphisms in the GABRA2 gene on the risk of alcoholism; (2) to determine associations between some genetic markers basing on haplotype analysis; Material and methods The study included 305 Caucasian males, among them 156 patients with ADS and 149 controls. These two groups, were compared as to the frequencies of haplotypes of the following polymorphisms: rs 279826, rs 279871, 279845 of the GABRA2 gene. All subjects were recruited in the North West region of Poland. Alcohol use and family history of alcoholism were assessed by means of a structured interview, based on the Polish version of Semi-Structured Assessment on Genetics in Alcoholism (SSAGA). Genomic DNA was extracted from venous blood. Polymorphisms were detected using PCR real time method. Conclusions Significant relationships documented by the haplotype analysis of ADS patients point to the usefulness of haplotype constructions in etiopathogenetic studies of ADS. The protective haplotype was G-G-A (p=0,01). In conclusion, larger – possibly multicentre – studies are needed to finally uncover the role of GABRA2 gene in determining ADS. Moreover, other candidate genes with a possible impact on the phenotype of alcohol-dependent individuals should be sought. Supported by grant of Ministry of Science and Higher Education (MNiSW no.: NN 402466540 ).

EPA-1081 - Case - control study of GABRA2 gene polymorphisms in alcohol dependence

Introduction Alcohol dependence (AD) is a highly prevalent disorder that is associated with serious morbidity and mortality. The development of alcohol dependence is the result of an interaction between environmental and hereditary factors. Previous studies have implicated GABRA2 receptor genes with AD, alcohol use patterns, and levels of response to alcohol. The aims of this study were to analyze the effects of polymorphisms in the GABRA2 gene on the risk of alcoholism ; associations between genotypes and alleles were studied; Material and methods This study included a group of 161 Caucasian subjects, with no history of psychiatric disorders other than alcohol or nicotine dependence as classified by ICD-10. The control group comprised 150 unrelated individuals matched for ethnicity and gender, and excluded for mental disorders using the Primary Care Evaluation of Mental Disorders questionnaire. All subjects were recruited in the North West region of Poland. Alcohol use and family history of alcoholism were assessed by means of a structured interview, based on the Polish version of Semi-Structured Assessment on Genetics in Alcoholism (SSAGA). Genomic DNA was extracted from venous blood. Polymorphisms (rs 279826, rs 279871, rs279845) were detected using PCR real time method. Conclusions Significant associations for genotypes and alleles were observed in rs 279871 polymorphism (p=0,31). The advantage of this research is that the genetic analysis of GABRA2 gene rs 279871 polymorphism was conducted on quite large and, most importantly, homogenous subgroups of alcohol-dependent patients. Supported by grant of Ministry of Science and Higher Education (MNiSW no.: NN 402466540 ).