E. Kornetova

Tardive dyskinesia and DRD3, HTR2A and HTR2C gene polymorphisms in Russian psychiatric inpatients from Siberia.

BACKGROUND Pharmacogenetics of tardive dyskinesia and dopamine D3 (DRD3), serotonin 2A (HTR2A), and 2C (HTR2C) receptors has been examined in various populations, but not in Russians. PURPOSE To investigate the association between orofaciolingual (TDof) and limb-truncal dyskinesias (TDlt) and Ser9Gly (DRD3), -1438G>A (HTR2A), and Cys23Ser (HTR2C) polymorphisms in Russian psychiatric inpatients from Tomsk, Siberia. METHODS In total, 146 subjects were included. Standard protocols were applied for genotyping. TDof and TDlt were assessed with AIMS items 1-4 and 5-7, respectively. Two-part model, logistic and log-normal regression analyses were applied to assess different variables (e.g., allele-carriership status, age, gender, and medication use). RESULTS TDlt, but not TDof, exhibited an association with Ser9Gly and Cys23Ser (with 9Gly and 23Ser alleles exhibiting opposite effects). However, -1438G>A was not associated with TDof and Dlt. CONCLUSIONS This is the first pharmacogenetic report on tardive dyskinesia in Russians. Subject to further replication, our findings extend and support the available data.

Missense polymorphisms in three oxidative-stress enzymes (GSTP1, SOD2, and GPX1) and dyskinesias in Russian psychiatric inpatients from Siberia

Neuronal degeneration due to oxidative stress (OS) has been proposed as a mechanism for tardive dyskinesia (TD) pathogenesis. Cellular defense mechanisms against OS may involve detoxification enzymes (e.g., glutathione peroxidase‐1, GPX1; superoxide dismutase‐2, SOD2 [also commonly known as MnSOD]; and glutathione S‐transferase P1, GSTP1). Several pharmacogenetic studies have examined TD and OS in different ethnic groups, but not in Russians. Here we report the association between orofaciolingual (TDof) and limb‐truncal dyskinesias (TDlt) and polymorphisms of GSTP1 (Ile105Val), MnSOD (Ala‐9Val), and GPX1 (Pro197Leu) genes in 146 Russian inpatients from Siberia. We applied AIMS instrument to rate dyskinesias. Two‐part model analyses, logistic and multivariate parametric regressions were applied to assess the effects of different variables (e.g., genotype, age, gender, and medication use). Our analyses do not suggest that Pro197Leu (GPX1) is associated with TD. However, our analyses suggest that the 105Val‐allele of Ile105Val (GSTP1) may be associated with a lower risk and a severity of TDof and TDlt and that Ile105Val pharmacogenetics may be different in Slavonic Caucasians from that in American Caucasians. Furthermore, we find evidence for an association between Ala‐9Val (MnSOD) and TDof, but not TDlt. Subject to further replication, our findings extend the available knowledge on the pharmacogenetics of TD and oxidative stress. Copyright

Prolactin gene polymorphism (− 1149 G/T) is associated with hyperprolactinemia in patients with schizophrenia treated with antipsychotics

BACKGROUND Antipsychotic drugs can cause hyperprolactinemia. However, hyperprolactinemia was also observed in treatment-naive patients with a first schizophrenic episode. This phenomenon might be related to the role of prolactin as a cytokine in autoimmune diseases. Extrapituitary prolactin production is regulated by an alternative promoter, which contains the functional single nucleotide polymorphism -1149 G/T (rs1341239). We examined whether this polymorphism was associated with hyperprolactinemia in patients with schizophrenia. METHOD We recruited 443 patients with schizophrenia and 126 healthy controls. The functional polymorphism -1149 G/T (rs1341239) in the prolactin gene was genotyped with multiplexed primer extension, combined with MALDI-TOF mass spectrometry. Genotype and allele frequencies were compared between groups with the χ2 test and logistic regression models adjusting for covariates. RESULTS The frequency of genotypes and alleles in patients with schizophrenia did not differ from those in control subjects. A comparison between patients with schizophrenia with and without hyperprolactinemia revealed significantly higher frequency of the G allele in patients with hyperprolactinemia than in patients without it (χ2=7.25; p=0.007; OR=1.44 [1.10-1.89]). Accordingly, patients with hyperprolactinemia carried the GG genotype more frequently than patients without hyperprolactinemia (χ2=9.49; p=0.009). This association remained significant after adjusting the estimates for such covariates as sex, age, duration of the diseases and the dose of chlorpromazine equivalents. CONCLUSION This study revealed a significant association between the polymorphic variant rs1341239 and the development of hyperprolactinemia in patients with schizophrenia. The serum prolactin concentration in patients with schizophrenia treated with antipsychotics may provide an indication of the activity of the gene that regulates extrapituitary prolactin production which is believed to play a role in the immune system.

Potential biomarkers of tardive dyskinesia: A multiplex analysis of blood serum

Potential biomarkers of tardive dyskinesia: a multiplex analysis of blood serum A.S. Boiko(1), E.G. Kornetova(2), S.A. Ivanova(1), A.J.M. Loonen(3) (1)Mental Health Research Institute, Tomsk National Research Medical Center of the Russian Academy of Sciences, Laboratory of Molecular Genetics and Biochemistry, Tomsk, Russia (2)Mental Health Research Institute, Tomsk National Research Medical Center of the Russian Academy of Sciences, Department of Endogenous Disorders, Tomsk, Russia (3)University of Groningen, Groningen Research Institute of Pharmacy, Groningen, The Netherlands Long-term antipsychotic treatment of schizophrenia is associated with the emergence of tardive dyskinesia (TD), a motor syndrome consisting of involuntary and hyperkinetic movements [1]. Pathogenesis of this drug-induced movement disorder is not yet fully established, but may be connected to oxidative stress-related indirect pathway neurotoxicity [2]. Dysregulations in immune, hormonal and neurotrophic systems have been postulated to be one of the mechanisms underlying this form of neurotoxicity [3,4]. Principle aims of translational psychiatric research are searching for biomarkers which can be used to diagnose pathological biochemical processes and to identify molecular targets for treatment as well as development of pharmacogenetic approaches to personalize this therapy. The aim is to study potential endocrine, neurotrophic and immunological markers of tardive dyskinesia in the blood serum of patients with schizophrenia with antipsychotic therapy. Methods: After obtaining approval of the study protocol by the local ethical committee, suitable participants were recruited from psychiatric hospitals. All subjects gave informed consent after proper explanation of the study. TD was assessed cross-sectionally by the use of the Abnormal Involuntary Movement Scale (AIMS) [1,5]. The concentrations of cortisol, brain-derived neurotrophic factor (BDNF), prolactin, cytokines (tumor necrosis factor (TNFa), interleukin 1 (IL-1β), interleukin 3 (IL-3), interleukin 6 (IL-6), interferon gamma (INF-γ) and S100β were measured in blood serum using the MILLIPLEX® MAP panels (Merck, Darmstadt, Germany) by the multiplex analyzer MAGPIX (Luminex, USA). Statistical analyses were performed using SPSS software for Windows. Results were expressed as median and quartile intervals (Me [Q1; Q3]) or mean and standard deviation (M ± SD). Differences were considered significant at p ≤ 0.05. Results: In total 180 patients with schizophrenia, 128 males and 52 females (age 39.2 ± 12.1 years), receiving long-term antipsychotic treatment were included. These patients were divided into two groups: 71 patients with tardive dyskinesia and 109 patients without this movement disorder. A significant (p = 0.04) decrease in BDNF concentration was observed in patients with TD (1.9 [1.01; 2.99] ng/ml) in comparison to patients without TD (2.66 [1.29; 3.89] ng/ml). An increase (p = 0.05) of the serum IL-6 level of patients with TD (5.69 [3.55; 7.4] pg/ml) was detected relative to patients without TD (4.69 [2.82; 6.13] pg/ml). In addition, a statistical trend (p = 0.06) of increased serum S100β concentration was found in TD patients (85.29 ± 5.53 ng/L) compared to patients without this side effect (75.14 ± 2.81 ng/L). No other significant differences were established concerning the other assayed biomarkers. Conclusions: The biological processes that might play a role in the development of TD are not confined to the human brain per se. Hormonal and immune systems are also involved, which may be related to these systems being closely interrelated. Furthermore, these parameters may provide information about risk factors of the movement disorder. Identifying markers that can be used as diagnostics or predictors of treatment response in people with tardive dyskinesia will be an important step towards being able to provide personalized treatment. References [1] Loonen, A.J., van Praag, H.M., 2007. Measuring movement disorders in antipsychotic drug trials: the need to define a new standard. J Clin Psychopharmacol 27, 423–430. [2] Loonen, A.J.M., Ivanova, S.A., 2013. New insights into the mechanism of druginduced dyskinesia. CNS Spectrums 18 (01), 15–20. [3] Wu, J.Q., Chen, D.Ch., Tan, Yu.L., Tan, Sh.P., Hui, L., Lv, M.H., Soares, J.C., Zhang, X.Y., 2015. Altered BDNF is correlated to cognition impairment in schizophrenia patients with tardive dyskinesia. Psychopharmacology 232, 223–232. [4] An, H.M., Tan, Y.L., Shi, J., Wang, Z.R., Soars, J.C., Wu, J.Q., Yang, F.D., Huang, X.F., Zhang, X.Y., 2015. Altered IL-2, IL-6 and IL-8 serum levels in schizophrenia patients with tardive dyskinesia. Schizophr Res 162 (1–3), 261–268. [5] Loonen, A.J.M., Doorschot, C.H., van Hemert, D.A., Oostelbos, M.C., Sijben, A.E., 2001. The schedule for the assessment of drug-induced movement disorders (SADIMoD): inter-rater reliability and construct validity. Int J Neuropsychopharmacol 4, 347–360. Keywords: Biological markers Schizophrenia: clinical Neuroleptics & antipsychotics: clinical

Gene polymorphism of serotonin receptors and drug-induced hyperprolactinemia in schizophrenic patients

Background: Antipsychotics have been used for more than 50 years in the treatment of schizophrenia and many other psychiatric disorders. Schizophrenic patients have to take antipsychotic medication for a long time, often throughout the life [1]. Hyper prolactinemia in these patients is a common and serious side effect of therapy with atypical antipsychotics. Genes coding for serotonin receptors are considered as candidate genes responsible for the particular antipsychotic effects of neuroleptics [2]. The present study aimed to investigate the role of polymorphisms of the serotonin receptors genes (HTR1A, HTR1B, HTR2A, HTR2C, HTR3A, HTR3B, HTR6) in the pathogenesis of antipsychotic-induced hyperprolactinemia in patients with schizophrenia. Methods: 446 Russian patients with schizophrenia were examined, including 225 women and 221 men. The average age of patients was 42.1±1.4 years. Evaluation of serum prolactin performed by ELISA using reagents set PRL Test System (USA). Genotyping was carried out on 29 polymorphic variants of the serotonin receptor genes HTR1A (rs6295, rs1364043, rs10042486, rs1800042, rs749099), HTR1B (rs6298, rs6296, rs130058), HTR2A (rs6311, rs6313, rs6314, rs7997012, rs1928040, rs9316233, rs2224721, rs6312), HTR2C (rs6318, rs5946189, rs569959, rs17326429, rs4911871, rs3813929, rs1801412, rs12858300), HTR3A (rs1062613, rs33940208, rs1176713), HTR3B (rs1176744) and HTR6 (rs1805054). The SPSS software was used for statistical analysis. The Hardy-Weinberg equilibrium (HWE) of genotypic frequencies was tested by the chi-square test. Results: We studied the association between polymorphisms of relevant 5-HT receptor genes and antipsychotic drug-induced hyperprolactinemia in patients with schizophrenia from Siberia. All patients with schizophrenia were divided into two groups: those with and without hyperprolactinemia. Statistically significant results were obtained for polymorphic variants of the genes rs6312 HTR2A (χ2 = 4.685; p = 0.030), rs12858300 HTR2C, rs569959 HTR2C, which suggests the participation of these polymorphic variants in the development of hyperprolactinemia. When men and women were studied separately, only rs569959 HTR2C (χ2 = 6.284; p = 0.043) in men reached the nominal significance threshold. This finding is probably related to the X-bound character of HTR2C (i.e., men are hemizygotes). Also, we found a statistically significant association rs12858300 HTR2C (χ2 = 9.429; p = 0.002) in the female group. We did not find statistically significant association for other polymorphisms HTR1A (rs6295, rs1364043, rs10042486, rs1800042, rs749099), HTR1B (rs6298, rs6296, rs130058), HTR2A (rs6311, rs6313, rs6314, rs7997012, rs1928040, rs9316233, rs2224721), HTR2C (rs6318, rs5946189, rs17326429, rs4911871, rs3813929, rs1801412), HTR3A (rs1062613, rs33940208, rs1176713), HTR3B (rs1176744) and HTR6 (rs1805054). Though hyperprolactinemia is primarily attributed to blockade of DRD2 in the pituitary gland, the secretion of prolactin is also modulated by 5-HT. The involvement of the HTR1A, HTR2A, HTR2C, and HTR3 in the serotonergic-induced prolactin response is well documented. Conclusion: Our results indicate that genetic variants of HTR2C may have functional consequences on the modulation of prolactin secretion. Further search for genetic markers associated with the development of side effects of neuroleptic therapy, will contribute to the development of effective methods of diagnosis, correction and treatment of disease, as well as of adherence of patients with mental disorders to psychotropic therapy.

1585 – Social adaptation and immune reactivity in schizophrenia

Study of social-clinical and biological components of adaptation in schizophrenia currently is of relevance. Objective Identification of interrelationship immune reactivity with social adaptation types of schizophrenic patients. Methods We examined 38 schizophrenic patients (F20.00, F20.01, F20.02). The scale SASS (Bosc M. et al., 1997) was used to estimate the social adaptation of schizophrenic patients. The immunological examination included the definition of phenotypes of surface receptors of immunocompetent cells, immunoglobulins, circulating immune complexes and phagocytosis levels. The research was carried out in two points: first - at admission, second - in 6 weeks of treatment. Results Three adaptation types have been revealed: normal type of social adaptation (15 patients, 39,47%), difficult social adaptation (18 persons), disadaptation (5 patients). By week 6 of treatment number of patients with normal social adaptation has heightened by 1,4 times (21 patients, 55,26%), improvement of indices of social adaptation was observed in 10 patients, type of social adaptation did not change in 28 patients. At admission T-immunodeficiency (CD2 + , CD3 + , CD16 + -lymphocytes decrease), increase CD8 + , HLADR + , CD20 + - lymphocytes, IgM and phagocytosis decrease were diagnosed among schizophrenic patients. During therapy, features of psychoneuroimmunomodulation were as follows: the normal type of social adaptation is relatively favorable in the immunological context. Patients with low level of social adaptation were characterized by clinical-immunological resistance toward therapy with deepening of T-cellular immune deficiency. Improvement of indices of social adaptation was accompanied by positive dynamic of immunological parameters. Conclusion Thus, social differences between groups have been confirmed by data according to immune reactivity.

Interaction between polymorphisms of oxidative enzymes and susceptibility to tardive dyskinesia in psychiatric patients

Tardive dyskinesia (TD) is a severe and potentially irreversible antipsychotic-induced movement disorder that has been and continues to be a significant problem associated with long-term use of antipsychotics [1]. About 30% of patients chronically exposed to neuroleptics may exhibit tardive dyskinesia. Neuronal degeneration due to oxidative stress has been proposed as a mechanism for the pathogenesis of tardive dyskinesia [2]. Several studies have examined the pharmacogenetics of TD and oxidative-stress enzymes in various ethnic groups, but not in Russians. Purpose: To investigate the possible effects of three missense polymorphisms (Ile105Val, Ala9Val, and Pro197Leu) in three oxidative-stress enzymes (GSTP1, SOD2, and GPX1, respectively) on orofaciolingual (TDof) and limb-truncal (TDlt) dyskinesias in Russian psychiatric inpatients from Siberia. Methods: After approval of the study protocol by the institutional bioethics committee, subjects were included from two psychiatric departments in Tomsk, Siberia. DNA extraction and TaqMan genotyping were conducted according to standard protocols and blind to the clinical status of the subjects. Subjects were characterized either as carriers or non-carriers of an allele. The sum of the first 7 items of the Abnormal Involuntary Movement Scale was utilized as a proxy for the severity of TD (TDsum). Multivariate parametric regression and two-part models were performed to identify the effect of different variables (allelecarriership status, age, gender, type of psychiatric department, use of anticholinergic and antipsychotic medication) on TD. Results: In total 146 Russian Caucasian patients (91 males, 55 females) with an age of 46.8±17.6 years (sample mean±SD) met the inclusion criteria. Seventy-nine (54%) subjects were included from a psychiatric department for permanently hospitalized, severely ill patients. The remaining 46% were less-severely ill inpatients from a psychiatric department for temporal hospitalization. About 95% of the patients had clinically-established schizophrenia (n = 138) and only 5% had schizotypical disorder (n = 8). The genotype distributions of Ile105Val (79 Ile/Ile, 53 Ile/Val, and 11 Val/Val), Ala9Val (37 Ala/Ala, 62 Ala/Val, and 42 Val/Val) and Pro197Leu (72 Pro/Pro, 60 Pro/Leu, and 11 Leu/Leu) were in consistency with the Hardy-Weinberg equilibrium. Our analyses do not suggest that Pro197Leu polymorphism (GPX1) is associated with TD. However, our analyses suggest that the 105Val-allele of Ile105Val (GSTP1) may be associated with a lower risk and a severity of TDof and TDlt and that Ile105Val pharmacogenetics may be different in Slavonic Caucasians from that in American Caucasians. This finding is plausible, since it has been suggested that Ile105Val polymorphism is functional with the 105Val-allele leading to a decreased enzymatic activity and detoxification capacity. Furthermore, we found evidence for an association between Ala9Val polymorphism (SOD2) and TDof, but not TDlt. Conclusions: This is the first study on the pharmacogenetics of TD and oxidative stress enzymes in Russian psychiatric subjects. Our findings extend the available knowledge on the pharmacogenetics of TD and oxidative stress.

Tardive dyskinesia and polymorphism of dopamine D3, serotonin 2A and 2C receptors in Russian psychiatric inpatients

Tardive dyskinesia (TD) is a potentially irreversible motor sideeffect occurring in about 30% of patients chronically exposed to neuroleptics. Genetic polymorphisms of dopamine D3 (DRD3), serotonin 2A (HTR2A) and 2C (HTR2C) receptors have been associated with TD. Currently, there are no pharmacogenetic studies describing the relationship between TD and polymorphisms of these genes in Russians. Purpose: To investigate the association between TD and Ser9Gly (DRD3), -1438G>A (HTR2A), and Cys23Ser (HTR2C) polymorphisms in Russian psychiatric patients. Methods: After approval of the study protocol by the institutional bioethics committee, subjects were included from two psychiatric departments in Tomsk, Siberia. DNA extraction and TaqMan genotyping were conducted according to standard protocols. Subjects were characterized either as carriers or non-carriers of an allele. The sum of the first 7 items of the Abnormal Involuntary Movement Scale was utilized as a proxy for the severity of TD (TDsum). Multivariate parametric regression and two-part models were performed to identify the effects of the different variables on TD. The following variables have been included in the analyses: allele-carriership status, age, gender, type of psychiatric department, use of anticholinergic and antipsychotic medication. Results: In total 146 Russian Caucasians patients (91 males, 55 females) with an age of 46.8±17.6 years (sample mean±SD) met the inclusion criteria. Seventy-nine (54%) subjects were included from a psychiatric department for permanently hospitalized, severely ill patients. The remaining 46% were less-severely ill inpatients from a psychiatric department for temporal hospitalization. About 95% of the patients had clinically-established schizophrenia (n = 138) and only 5% had schizotypical disorder (n = 8). The genotype distributions of DRD3 Ser9Gly (69 Ser9/Ser9, 67 Ser9/Gly9, and 9 Gly9/Gly9) and HTR2A-1438G>A (62 GG, 68 GA, and 14 AA) were in agreement with Hardy-Weinberg Equilibrium. The genotype distribution for HTR2C Cys23Ser (X-chromosomal) was: 116 Cys23/Cys23, 14 Cys23/Ser23 and 13 Ser23/Ser23. Parametric regression showed that carriership of the Gly9 (DRD3 Ser9Gly) and Ser23-alleles (HTR2C Cys23Ser) almost offer the highest explanatory power among all of the variables studied. Ser9Gly and Cys23Ser polymorphisms may however act in opposite directions. Whereas carriers of Gly9-allele may have 36% higher TDsum values (p = 0.022), carriers of Cys23-allele exhibited 30% lower TDsum values (p=0.035). Furthermore, two-part model analyses showed that carriership of Gly9 or-1438A-alleles (DRD3 and HTR2A genes, respectively) may be associated with 2.5-2.8 folds higher risk of having a TDsum > 0 (p=0.048 and p=0.026, respectively). Discussion: The allelic effects observed in the current study (Cys23-allele: protection against TD; Gly9 and-1438A-alleles: predisposition for higher TDsum values) are in agreement with the literature [1] and [2]. Conclusions: This is the first study on the pharmacogenetics of TD in Russian psychiatric patients. Subject to further replication, our findings reproduce and extend previously published pharmacogenetic data on TD.