Investigation of maternal effects, maternal-fetal interactions and parent-of-origin effects, using mothers and their offspring with schizophrenia

P.1.a. Basic and clinical neuroscience − Genetics and epigenetics An important influence of potassium channels on dopaminergic neurotransmission is well documented [2]. Studies show that PIP5K2A and SGK1 modulate the activity of neuronal potassium channels [3,4]. Antipsychotic effect of neuroleptics is thought to blockade the dopamine D2-receptors and to change dopaminergic neurotransmission, which in turn may cause extrapyramidal disorders, including tardive dyskinesia. It was previously shown an association between rs10828317 PIP5K2A and dyskinesia in schizophrenia [5]. Objective: The aim of our study was to investigate the association of polymorphisms of PIP5K2A (rs1417374, rs946961, rs1132816, rs11013052, rs943190, rs10430590, rs943194, rs1171506, rs11013052) and SGK1 (rs9373085, rs1743964, rs1743966, rs1057293, rs17063563, rs1009840) genes with druginduced dyskinesia in Russian schizophrenic patients. Materials and Methods: Blood samples were taken from 443 Russian Caucasian patients (61.4% male and 38.6% female) with a clinical diagnosis of schizophrenia (ICD-10: F20). The average age of patients 38±14.5 years; duration of the disease at the time of the survey 23±8.9 years. The drug-induced dyskinesia was assessed using standard international scale AIMS. Results: When comparing the group of patients with and without dyskinesia we did not find any associations between polymorphic variants of PIP5K2A and SGK1 genes and tardive dyskinesia. Additional comparison of the frequency distributions of genotypes and alleles were carried out in groups of men and women. In the group of men we identified the association of A allele (c2 = 3.80, p = 0.049) and AA genotype (c2 = 6.17, p = 0.046) of rs1057293 in SGK1 gene with drug-induced dyskinesia. In the group of women we identified the association of AA and GG genotypes of rs1743964 in SGK1 gene with dyskinesia (c2 = 6.006, p = 0.049). To assess clinical heterogeneity of tardive dyskinesia we subdivided group of patients with extrapyramidal disorders into groups of patients with orofaciolingual symptoms and patients with limb-truncal symptoms. It was found that orofaciolingual form was associated with CC genotype (c2 = 6.299, p = 0.043) and C allele (c2 = 5.11, p = 0.02) of rs11013052 PIP5K2A (in the group of men) and AA and CC homozygous genotypes of rs1009840 in SGK1 (c2 = 5.818, p = 0.049) (in the group of women). In the total group of patients we did not identify any gene polymorphism’s associations with clinically different forms of dyskinesia. This study shows that gene polymorphisms in men and women may differently affect the development of side effects of neuroleptic therapy. Conclusions: Thus, PIP5K2A and SGK1 genes are involved in the development of tardive dyskinesia induced by long-term therapy with neuroleptics and phenotypically different forms of tardive dyskinesia − orofaciolingual and limb-truncal − characterized by different genetic features. References [1] Loonen, A.J., van Praag, H.M., 2007. Measuring movement disorders in antipsychotic drug trials: the need to define a new standard. Journal of Clinical Psychopharmacology 27(5), 423–430. [2] Zhang, H., Rodgers, E.W., Krenz, W.D., Clark, M.C., Baro, D.J., 2010. Cell specific dopamine modulation of the transient potassium current in the pyloric network by the canonical D1 receptor signal transduction cascade. J Neurophysiol 104, 873–884. [3] Lang, F., Boehmer, C., Palmada, M., Seebohm, G., Strutz-Seebohm, N., Vallon, V., 2006. (Patho)physiological significance of the serum- and glucocorticoid-inducible kinase isoforms. Physiol Rev 86(4), 1151−78. [4] Fedorenko, O. Strutz-Seebohm, N., Henrion, U., Ureche, O., Seebohm, G., Lang, F.A., Lang, U.E., 2008. A schizophrenia-linked mutation in PIP5K2A fails to activate neuronal M channels. Psychopharmacology (Berl) 199, 47−54.

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[5] Fedorenko O.Y., Toshchakova V.A., Boyarko E.G., Semke A.V., Bokhan N.A., Ivanova S.A., Loonen A.J.M., Lang F., Govorin N.V., Aftanas L.I., 2015. Association study indicates a protective role of phosphatidylinositol-4-phosphate-5-kinase against tardivedyskinesia. The International Journal of Neuropsychopharmacology. 18(6), 1−6. Disclosure statement: The study was supported by a Russian Science Foundation No. 14−35–00023 “Laboratory of pharmacogenetic research on personalized therapy of mental and neurodegenerative disorders”

P.1.a.032 Investigation of maternal effects, maternalfetal interactions and parent-of-origin effects, using mothers and their offspring with schizophrenia B.D. Lee1 , D.K. Lee2 ° , H.C. Kim3 , W.Y. Jung4 1 Pusan National University Hospital, Department of Psychiatry, Busan, Republic of Korea; 2 Good gang-an General Hospital, Department of Psychiatry, Busan, Republic of Korea; 3 Kosin University Hospital, Department of Psychiatry, Busan, Republic of Korea; 4 Pusan National University Yangsan Hospital, Department of Psychiatry, Gyeongnam, Republic of Korea Background: Schizophrenia is a frequent and disabling disease associated with heterogeneous psychiatric phenotypes. It emerges during childhood, adolescence or young adulthood and has dramatic consequences for the affected individuals, causing considerable familial and social burden, as well as increasing health expenses. The prevailing hypothesis regarding the causation proposes that a combination of genetic and/or environmental factors during critical periods of brain development increases the risk for this illness. Introduction: Many complex genetic effects, including epigenetic effects, may be expected to operate via mechanisms in the inter-uterine environment. A popular design for the investigation of such effects, including effects of parent-of-origin (imprinting), maternal genotype, and maternal-fetal genotype interactions, is to collect DNA from affected offspring and their mothers (case/ mother duos) and to compare with an appropriate control sample. An alternative design uses data from cases and both parents (case/parent trios) but does not require controls. In this study, we describe a novel implementation of a multinomial modeling approach that allows the estimation of such genetic effects using either case/mother duos or case/parent trios. We investigate the effects of estimation of maternal, imprinting and interaction effects using multimodal modeling using parents and their offspring with schizophrenia in Korean population. Subjects and Methods: We have recruited 27 probands (with schizophrenia) with their parents and siblings whenever possible. We analyzed 96 SNPs (Single Nucleotide Polymorphisms) of 17 functionally only relevant genes and 21 neuronal genes relevant to schizophrenia for DNA samples that was checked for the data quality and genotype error. We used EMIM analysis program for the estimation of maternal, imprinting and interaction effects using multimodal modeling. Results: Of analyzed 96 SNPs functionally and positionally relevant to schizophrenia, very significant SNP (rs324420) was suggested in EMIM analysis for child genetics effects. (p = 1.5×10−4 ) rs 324420 was also significant for child genetic effects allowing for maternal genetic effects. (p = 5.3×10−4 ) with very stringent multiple comparison Bonferroni correction. Additionally, no significant SNP was presented in the analysis results for maternal genetic effects allowing for child genetic effects.

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P.1.a. Basic and clinical neuroscience − Genetics and epigenetics

Discussion: Epigenetics and gene-environment interactions are represented underlying statistical genetics. Our results are the pilot study for epigenetic study in mental disorder and help to understanding and use of EMIM statistical genetics analysis program with many limitations including small pedigree numbers. References [1] Sinsheimer, J.S., Palmer, C.G., Woodward, J.A., 2003. Detecting genotype combinations that increase risk for disease: maternal-fetal genotype incompatibility test. Genet Epidemiol. 24(1), 1−13. [2] Howey, R., Cordell, H.J., 2012. PREMIM and EMIM: tools for estimation of maternal, imprinting and interaction effects using multinomial modelling. BMC Bioinformatics. 27; 13: 149. [3] Howey, R., Mamasoula, C., T¨opf, A., Nudel, R., Goodship, J.A., Keavney, B.D., Cordell, H.J., 2015. Increased Power for Detection of Parent-of-Origin Effects via the Use of Haplotype Estimation. Am J Hum Genet. 3; 97(3): 419−34. [4] Wang, S., Yu, Z., Miller, R.L., Tang, D., Perera, F.P., 2011, Methods for detecting interactions between imprinted genes and environmental exposures using birth cohort designs with mother-offspring pairs. Hum Hered. 2011; 71(3): 196–208.

P.1.a.033 Transcriptional mechanisms mediated by cortisol treatment N. Cattane1 ° , N. Lopizzo1 , A. Borsini2 , C. Pariante2 , A. Cattaneo1,2 1 IRCCS Centro San Giovanni di Dio, Biological Psychiatry Unit, Brescia, Italy; 2 King’s College London, Institute of Psychiatry- Psychology and Neuroscience, London, United Kingdom Early life severe stress can lead to lifelong molecular changes in the form of epigenetic modifications that can shape and store the molecular response of a cell to its environment, providing a potential mechanism for gene-environment interactions [1]. Epigenetics subsumes mechanisms of functional control over the genetic information without changing DNA sequence and, among the most investigated there are: (i) DNA methylation predominantly at CG dinucleotides (CpG), which can influence the spatial structure of the DNA and the binding or the repression of specific DNAbinding proteins to the DNA [2] and (ii) histone modifications, which influence the condensation of the DNA around histone proteins and regulate the accessibility of functional regions to transcriptional regulators [3]. However, up to now, the contribution of each mechanisms on long term exposure to stress is not clear. Thus, the aim of this study was to identify the possible epigenetic mechanisms underlying the long lasting changes associated with early stressful life events by using an in vitro model represented by the immortalized multipotent human hippocampal progenitor cell line HPC03A/07 (ReNeuron Ltd., Surrey, UK) [4]. We treated these cells during the proliferation phase with high concentration of cortisol (100mM) and we then assessed the modulation of key genes involved in different epigenetic mechanisms. In particular, RNA samples were processed onto Human Gene 1.1 array strips (Affymetrix, Inc., Santa Clara, CA, USA) which cover 36,079 transcripts. Quality control and statistical analyses were performed using Partek Genomic Suite 6.6 software. We specifically looked at genes modulated by cortisol and involved in the epigenetic modification machinery. We did not observe modulation in genes responsible for the maintenance of DNA methylation, such as DNA (cytosine-5)-methyltransferase 1 (DNMT1, fold change (FC)=-1.1, p > 0.05), methyl CpG binding protein 2 (MeCP2, FC=-1.1, p > 0.05), the TET methylcytosine

dioxygenase proteins (TET1, FC=-1.1, p > 0.05; TET2, FC=-1.0, p > 0.05; TET3, FC=-1.2, p > 0.05) and the methyl-CpG-binding domain protein 2 (MBD2, FC = 1.0, p > 0.05). Interestingly, when we looked at genes involved in the histone-modifying cluster family, we found a significant modulation in the levels of several histone modifying enzymes, which were all down-regulated: for example, HIST1H2AB (FC=-1.75; p = 5.66 e-3), HIST2H2BF (FC=-1.31, p = 8.34 e-3) and HIST2H3D (FC=-2.08, p = 3.67 e-2). These enzymes are essential for the creation of the nucleosomes and allow the compaction of chromatin into higher order structures. These results suggest that early life stressful events can influence the transcriptional regulation of epigenetic writers, readers, and erasers and may alter the expression of genes critically involved in the epigenetic regulation. We are now investigating whether changes in these histone modifying enzymes can be maintained over time and/or whether changes in other epigenetic players can be manifested later in time by assessing cells, treated with cortisol during the proliferation phase, after a washout period of 20 days during the differentiation phase. The possibility to directly or indirectly interfere with this mechanism could lead to the identification of new pharmacological targets for preventive therapies to reverse or ameliorate the disease status. References [1] Klengel, T., Binder, E.B., 2015. Epigenetics of Stress-Related Psychiatric Disorders and Gene × Environment Interactions. Neuron. Jun 17; 86(6): 1343−57. doi: 10.1016/j.neuron.2015.05.036. [2] Slatkin, M., 2009. Epigenetic inheritance and the missing heritability problem. Genetics 182, 845–850. [3] Levine, A., et al., 2012. Early life stress triggers sustained changes in histone deacetylase expression and histone H4 modifications that alter responsiveness to adolescent antidepressant treatment Neurobiol. Dis. 45, 488–498. [4] Anacker, C., et al., 2012. Glucocorticoid-related molecular signaling pathways regulating hippocampal neurogenesis. Neuropsychopharmacology. 2013 Apr; 38(5): 872−83. doi: 10.1038/npp.253.

P.1.a.034 MicroRNA regulation of candidate genes for Tourette syndrome L. Pagliaroli1 ° , A. Vereczkei1 , C. Barta1 1 Semmelweis Univesity, Institute of Medical Chemistry- Molecular Biology and Pathobiochemistry, Budapest, Hungary Background: Tourette Syndrome (TS) is a neurodevelopmental disorder that presents early in childhood and is marked by the appearance of multiple involuntary motor tics and at least one vocal tic. It presents high comorbidity rates with other disorders such as attention deficit hyperactivity disorder and obsessive compulsive disorder. Despite a strong genetic contribution, the molecular mechanisms behind TS are still uncertain, although multiple lines of evidence suggest involvement of specific candidate genes and corresponding epigenetic mechanisms via miRNA regulators. To date only a few genetic findings have been replicated in TS. Among these the nicotinic acetylcholine receptor alpha 7 subunit (CHRNA7) gene has been recently suggested as candidate susceptibility gene. CHRNA7 is known to regulate a wide variety of developmental and secretory functions [1], however the mechanism of its transcriptional regulation is still unclear. Another promising finding of the last years in TS genetic research is Netrin 4 (NTN4), which belongs to a family of extracellular proteins that direct axon outgrowth and guidance [2]. Several microRNAs have already been associated with neuropsychiatric disorders, but