Gohar Mkrtchyan

Markers of apoptotic dysfunctions in schizophrenia

According to the modern concepts, alterations of apoptosis and its genetic regulation are associated with the etiopathogenesis of schizophrenia, which is observed at both the brain and peripheral blood levels. However, studies of this phenomenon are at the initial stage, and the molecular and cellular mechanisms that underlie the anomalies of the processes of apoptotic cell death in schizophrenia are unclear. In the present study, we determined the levels of apoptotic markers, annexin A5 and H-ficolin proteins, in the sera of patients with chronic and first-episode schizophrenia and healthy subjects to test the proposed relationship between schizophrenia and the rs11575945 (−1C/T) single-nucleotide substitution (functional polymorphism) of Kozak consensus sequence in the regulatory region of the annexin A5 gene. Methods of a solid-phase enzyme-linked immunosorbent assay and polymerase chain reaction with allele-specific primers were used. It was shown that the pathogenesis of schizophrenia is characterized by an increased rate of apoptosis, which is more pronounced in the case of the first-episode neuroleptic-free patients than in the case of chronic patients that receive typical neuroleptic haloperidol. It was also shown that the rs11575945 polymorphism of the annexin A5 gene is associated with schizophrenia, and its minor allele is responsible for higher levels of the annexin A5 protein in the blood and represents one of the risk factors for the development of this disease.

Increased levels of circulating Annexin A5 in Familial Mediterranean fever

BackgroundFamilial Mediterranean fever is a genetic autoinflammatory disease most commonly affecting the ethnic groups originating from around the Mediterranean Sea. Apoptosis plays an important role in down-regulation of the inflammatory response by reducing the lifespan of activated immunocompetent cells. Thus, increased apoptosis may be associated with pathogenesis of familial Mediterranean fever.MethodsIn the present study we determined the serum levels of apoptotic marker, Annexin A5, in familial Mediterranean fever patients, within an attack and attack-free, in comparison to healthy subjects and assessed the influence of colchicine treatment on this parameter. In addition, in all study subjects serum levels of C-reactive protein and interleukine-1β, and the total leukocyte count were also determined.ResultsOur results demonstrated that pathogenesis of familial Mediterranean fever is characterized by the increased levels of circulating Annexin A5, which is higher in patients within the attack and which associate with the increased levels of C-reactive protein and interleukine-1β and total leukocyte count.ConclusionsThe results obtained indicate elevated rates of apoptosis of subpopulations of leukocytes involved in autoinflammation and recurrent episodes of fever in familial Mediterranean fever. It was also revealed that regular colchicine treatment sufficiently decreases the rate of apoptosis in familial Mediterranean fever patients by affecting the intensity of autoinflammatory reactions.

Involvement of Anomalous Apoptosis in Impairments to Synaptic Plasticity in Post-Traumatic Stress Disorder

Enzyme-linked immunosorbent assay was used to measure levels of apoptosis and synaptic plasticity marker proteins, i.e., annexin A5 and complexin 2 respectively, as well as the proinflammatory cytokine tumor necrosis factor α (TNF-α), in serum from patients with post-traumatic stress disorder (PTSD) in comparison with healthy subjects. Correlations between these parameters were studied. The results obtained here showed that annexin A5 and complexin 2 concentrations in PTSD patients were significantly lower than normal, while TNF-α levels were higher. PTSD patients showed a positive correlation between annexin A5 and complexin 2 levels on the one hand, and a negative correlation between annexin A5 and TNF-α levels on the other. These data lead to the conclusion that the pathogenesis of PTSD is characterized by reduced apoptosis associated with defects in synaptic plasticity. It is suggested that anomalous apoptosis may also be among the factors supporting the development of the chronic inflammation typical of the pathogenesis of PTSD.

Schizophrenia-associated Risk and Protective Variants of c-Fos Encoding Gene

Defects in synaptic plasticity play a key role in pathophysiology of schizophrenia. Pathomechanisms responsible for synaptic plasticity alterations in schizophrenia are very complicated and not well defined. Transcription factor c-Fos plays an important role in regulation of synaptic plasticity. In the present study we evaluated the association of rs7101 and rs1063169 single nucleotide polymorphisms (SNPs) of c-Fos encoding gene (FOS) with schizophrenia. A total of 604 DNA samples of schizophrenia-affected and healthy subjects of Armenian ancestry were genotyped using polymerase chain reaction with sequence-specific primers. Also, comparative determination of the blood levels of c-Fos protein in schizophrenia patients and controls was performed using the enzyme-linked immunosorbent assay. Potential interaction between protein level and genotypes as well as relationships between genotypes/protein level and clinical-demographic characteristics of schizophrenia patients were assessed. The results obtained demonstrated that mutant allele of FOS rs1063169 SNP is negatively associated with schizophrenia and may be nominated as a protective factor for this disorder. On the other hand, according to our results, the FOS rs7101T mutant allele is positively associated with schizophrenia and, therefore, may be considered as a risk factor for this disorder. In addition, decreased c-Fos plasma levels in schizophrenia patients compared to controls were found. In conclusion, the results of this study suggest that FOS is among the candidate genes of schizophrenia and that changes in the expression of c-Fos protein may contribute to molecular pathomechanisms of schizophrenia-related alterations in synaptic plasticity.

Alterations in the Immune Response, Apoptosis and Synaptic Plasticity in Posttraumatic Stress Disorder: Molecular Indicators and Relation to Clinical Symptoms

Posttraumatic stress disorder (PTSD) (ICD-10 codes: F43.1, F62.0; DSM-IV-TR code: 309.81) [1, 2] is a complex severe and chronic psychiatric illness influenced by environmental and genetic factors [3-10]. PTSD is an anxiety disorder developed in a person experiencing, wit‐ nessing, or learning about an extreme physically or psychologically distressing event, asso‐ ciated with unprecedented violence [11, 12]. Traumatic events that can trigger PTSD include massacres, mass murder scenes, international, civil, political, ethnic and religious wars, gen‐ ocides, natural and man-made disasters, criminal assaults, serious accidents, terrorist at‐ tacks, incarceration, trafficking, rape and other types of sexual assaults [12-17], life threatening illness and the sudden death of a loved one, serious medical illness, injury, sur‐ gery, hostage, kidnapping, difficult labors, etc [18-20]. Individuals who experience a trauma of this nature may develop symptoms that fall into three distinct clusters: re-experiencing phenomenon; avoidance and numbing; and autonomic hyperarousal. Symptoms usually be‐ gin within the first 3 months after the traumatic event and last for many years, although there may be a delay of months, or even years, before symptoms appear. PTSD patients are characterized by severe emotional state, sharp reduction in adaptive and information receiv‐ ing abilities. They usually remain out of society, become drug addicted, alcoholic and often commit suicide [21-24]. Degrees of risk to develop PTSD from different traumatic events are presented in table 1.

L-Ficolin and H-Ficolin as Links Between Upregulated Immune Response and Increased Rate of Apoptosis in Familial Mediterranean Fever

Familial Mediterranean fever (FMF) is autoinflammatory disease characterized by self-limited recurrent episodes of fever (attack) and serosal inflammation. Recent findings suggest the involvement of aberrant apoptosis in pathomechanisms of this disorder. FMF is caused by mutations in the gene of pyrin, a protein, which controls inflammation and apoptosis by regulating interleukin-1I² (IL-1I²) processing and nuclear-factor-IoB and pro-caspase-1 activation. L-ficolin and H-ficolin participate in the clearance of apoptotic cells acting as opsonins or by activating the complement lectin pathway. In this study we evaluated blood serum levels of L- and H-ficolins, and IL-I² by the enzyme-linked immunosorbent assay and total leukocyte count (TLC) in attack and attack-free FMF patients in comparison to healthy subjects. Correlation between measured parameters was assessed. Data was analyzed by parametric and nonparametric statistics. Elevated H-ficolin level and TLC were detected in patients during attack period, whereas increased levels of L-ficolin and IL-1I² were found in both attack and attack free patients with higher values during attack. Positive correlation between H-ficolin and L-ficolin levels in patients and healthy subjects was detected. Our results suggest excess production of L- and H-ficolins and increased apoptosis rate in FMF and indicate that H-ficolin is operating during development of acute autoinflammatory reactions, whereas L-ficolin is operating in both acute and subclinical autoinflammatory responses associated with this disease.

394 – Implication of genetic polymorphisms and changes in expression levels of proteins regulating neuronal plasticity and apoptosis in schizophrenia disorder

Introduction Pathologic changes in neuronal plasticity and apoptosis are important factors influencing alterations in synaptic transmission and impaired cognitive function, characteristic features of schizophrenia. Brain derived neurotrophic factor (BDNF), complexin-2, and annexin A-5 are known as members of regulatory pathways responsible for maintenance of neuronal plasticity and apoptosis. Objectives The study was focused on assessment of possible association of functional single nucleotide polymorphisms (SNPs) and expression levels of BDNF, complexin-2, and annexin A-5 with schizophrenia. Aims For this purpose we investigated functional polymorphisms and plasma levels of these proteins in schizophreniaaffected neuroleptic-treated and neuroleptic-free patients and healthy controls. Methods DNA samples were genotyped by polymerase chain reaction with sequence-specific primers. Plasma levels of BDNF, complexin-2, and annexin A-5 were measured with an enzyme-linked immunosorbent assay. The significance of differences between allele and phenotype frequencies in study groups was determined using Pearsons χ2 test. For evaluation of intergroup differences in the levels of BDNF, complexin-2, and annexin A-5, both parametric and non-parametric statistics were used. The assessment of correlation between the measured parameters within each group was also performed. P-values less than 0.05 were considered significant. Results In patients with schizophrenia, as compared to controls, the carriers of mutant alleles of the functional polymorphisms of BDNF, complexin-2, and annexin A-5 genes were overrepresented, and the blood levels of these proteins were altered. Conclusions Functional polymorphisms of genes encoding BDNF, complexin-2, and annexin A-5 proteins are associated with schizophrenia resulting in altered expression levels of these proteins upon this disorder.