1. Polymorphisms in the CRP and C1 Q genes and schizophrenia in Armenian population: A pilot study Zakharyan R 1,2, Khoyetsyan A 1, Chavushyan A 1, Arakelyan A 1, Boyajyan A 1, Stahelova A 2, Mrazek F 2, Petrek M 2,3 1 Laboratory of Macromolecular Complexes, Institute of Molecular Biology, Yerevan, NAS Armenia 2 Laboratory of Immunogenomics and Proteomics, Dept. of Immunology, Medical Faculty, Palacky University, Olomouc 3 Div. of Clinical Biochemistry and Immunogenetics, Faculty Hospital, Olomouc

2. Schizophrenia Schizophrenia is a complex mental disease with genetic component characterized by behavioral, cognitive and social abnormalities According to data provided by the World Health Organization (WHO, 2008) this disease affects about 7 per thousand of the adult population, mostly in the age group 15-35 years (first episodes) Disease heritability is about 80%, siblings recurrence risk is 10%

3. CRP  C1Q in schizophrenia CRP is an acute and chronic phase inflammation marker. C1Q is the first and key component of classical activation pathway of complement and consists of 3 subunits –C1QA, C1QB, C1QC. The CRP (Hakobyan et al, 2005; Dickerson et al. 2007) and C1Q (Boyajyan et al, 2008) are upregulated in schizophrenia, and likely contribute to disease progression Hypothesis The functional variants of the CRP and C1Q genes might be involved in pathogenesis of schizophrenia.

4. Objective: To investigate possible association of selected variants in CRP and C1Q genes with susceptibility to schizophrenia. Selected SNP variants: CRP: rs1417938 (T/A) – intron rs1800947 (C/G) – intron rs1205 (C/T) – UTR-3´ C1QB: rs291982 (G/T) – intron rs631090 (T/C) – intron

5. Subjects and methods Study population ◦ patients with paranoid schizophrenia n=103 ◦ healthy subjects n=105 ◦ all subjects unrelated Armenians Methods ◦ DNA extraction: Standard phenol-chloroform method using 3-5ml venous blood from subjects of both groups ◦ Genotyping: Polymerase chain reaction with sequence- specific primers (PCR-SSP) ◦ Statistical analysis: Pearson’s Chi-square test was performed for analysis of genotyping data.

6. Proportion of mutant allele frequencies Results: CRP rs1417938 rs1800947 rs1205

7. Results : CRP rs1417938 P=0.1 rs1205 P=0.1

8. Results : CRP The distribution of genotypes for all 3 CRP SNPs corresponded to the Hardy-Weinberg equilibrium No association between the selected three SNPs in the CRP gene and schizophrenia has been found (P>0.05).

9. Results: C1 QB Proportion of mutant allele Pn=0.01; OR=0,46 0.5346<CI<0.9084 rs291982 ControlSchizophrenia

10. Results : C1 QB The distribution of genotypes for both C1QB SNPs corresponded to the Hardy-Weinberg equilibrium The carriers of the C1QB rs291982*T allele were significantly less frequent in patients with schizophrenia compared to control subjects (P nominal =0.01, P corr =0.05) No association between the C1QB rs631090 SNP and schizophrenia has been found.

11. Conclusions Polymorphism rs291982 of the C1QB gene is associated with schizophrenia in this pilot study. C1QB gene variants or a locus located nearby may therefore be involved in susceptibility to schizophrenia. No association between the C1QB rs631090 SNP and schizophrenia has been found. No association between the selected variants of the CRP gene and schizophrenia was observed.

12. Discussion To our knowledge, this is the first study investigating a relationship of C1Q gene with schizophrenia. According to the recommendation for the genetic – association studies our results should be replicated in independent cohorts. The exact role of C1 Q molecules and their gene variants in schizophrenia remains to be clarified.

13. Future plans To enlarge study group for gaining more statistical power and for subanalysis according to the particular disease phenotypes To evaluate several other CRP polymorphisms to achieve more coverage of the gene To investigate additional two SNPs within C1Q genes (rs913243 and rs172378) using PCR techniques To perform haplotype mapping for C1Q genes

14. Roksana Zakharyan Presenting author: Roksana Zakharyan PhD Student & Junior researcher Institute of Molecular Biology, Yerevan, Armenia Acknowledgements Study is supported by International Visegrad Fund Prof. Anna Boyajyan, DrSc Yeravan, Armenia Prof. MUDr. Martin Petrek, CSc & MUDr. Frantisek Mrazek, PhD. Olomouc, Czech Republic

15. Thank you