Distinct Serum Protein Pattern in Paranoid Schizophrena

Distinct Serum Protein Pattern in Paranoid SchizophrenaA DISTINCT blood blood serum PROTEIN PATTERN IN PATIENTS WITH PARANOID SCHIZOPHRENIAA1N. A. Timofeyeva1,3, I. V. Alekseeva1,3, S. A. Ivanova2,4, G. G. Simutkin2, A. V. Semke2, I. S. Losenkov2, N. A. Bokhan2, O. S. Fedorova1,3, A. A. Chernonosov1,3*1 Institute of Chemical Biology and Funda intellectual medication SB RAS, Novosibirsk 630090, Russia2 Mental Health Research Institute, Tomsk bailiwick Research Medical Center of the Russian Academy of Sciences, Tomsk, Russia3 Novosibirsk State University, Novosibirsk, Russia 4 National Research Tomsk Polytechnic University, Tomsk, RussiaABSTRACTThe proteomic approach, namely, a combination of 2D mousse electrophoresis and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry, is a powerful tool that allows lookers to identify proteins that be differentially expressed in disease states. dementia praecox is a chronic mental illness, whose aetiology is stil l unclear therefore, information about differences in serum protein patterns may improve the understanding of the pathophysiology of schizophrenia.The goal of this field of study was to use the proteomic approach to identify modify protein levels in the serum samples from patients with schizophrenia. For this study, blood was collected from 10 patients with paranoid schizophrenia and 10 healthy volunteers. We uncovered major changes in the expression of such(prenominal) proteins as apolipoproteins of classes A4 and C3, transthyretin (TTR), and serum amyloid A1. Further more than(prenominal), an change magnitude in expression was found only for apolipoprotein A4, whereas the expression of apolipoprotein C3, TTR, and serum amyloid A1 was comed. The discovered differences in the expression of serum proteins (TTR and serum amyloid) ar in good agreement with the results obtained by other seek groups during analyses of cerebrospinal fluid or post-mortem brilliance tissues by ot her methods.Key haggle psychiatric disorder, schizophrenia, A2proteomics, 2D electrophoresis, MALDI-TOF mass spectrometry, biomarker, serum.IntroductionAlthough in recent years, great progress has been made in reducing mortality and in the treatment of common illnesses such as cancer and cardiovascular disease, the mortality caused by mental disorders remains same(predicate) 1. Schizophrenia is a chronic mental illness, whose aetiology is still unclear. Schizophrenia is characterised by hallucinations, delusions (positive psychotic symptoms), affective problems A3(negative psychotic symptoms), and cognitive dysfunction 2. A number of hypotheses have been proposed about the pathogenesis of schizophrenia, for example, aberrations A4in the pathways of transmission of neurotransmitters dopamine and serotonin 3, 4 or pathological changes in embryonic neurogenesis owing to variations in gene neuregulin-1 5, as well as oxidative-stress-mediated cell damage cod to lowered levels of antio xidant defence enzymes in patients with schizophreniaA5 6. Such pathogenesis may be caused by a dysfunction of some enzymes (proteins) as well as changes of their quantity in the blood of these patients.At the same time, there is no information about differences in serum protein patterns that can be used for typing of psychopathologies among individuals at luck of developing psychiatric disorders 7. Diagnosis and nosology rely on symptoms and accumulated clinical observations, and thus far, have been based mostly on interviews with patients and on patients indwelling complaints 8. Moreover, menstruum medications still have substantial adverse effects and/or require weeks for therapeutic effects to manifest themselves non all patients respond to current pharmacotherapy 9. In sum, an insufficient understanding of psychiatric disorders at the molecular level and the lack of disease-specific changes in serum protein patterns prevent optimisation of diagnosis and treatment of such com plex psychiatric disorders as schizophreniaA6.The proteomic approach, namely, the combination of 2D gel electrophoresis and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS), allows researchers to reliably identify proteins degage from human bodily fluids 10, 11. Our exploratory study 12 showed that 2D gel electrophoresis is suitable for isolation of proteins from blood of patients with mental disorders. ThereforeA7, in the puzzle work, clinical blood samples from patients with a diagnosis of schizophrenia were tested to identify quantitative differences in the proteomic profile of serum.Results and DiscussionIn this study, we examined blood serum of healthy citizenry and patients with paranoid schizophrenia to search for quantitative and/or qualitative differences in proteins associated with this mental disorder. The use of 2D gel electrophoresis enables researchers to simultaneously isolate more than 300 protein spots on one gel contain ing 150 g of protein for subsequent MALDI-TOF MS/MS analysis 10, 11. We analysed differences in serum protein patterns by compargon the gels between the patients and healthy controls. The analysis A8of protein patterns in serum was focused on those protein spots that differed in 2D gels between the patients and healthy controls. Such protein spots were analysed by means of the Gel-Pro Analyzer software and normalised to the sum of three proteins (a, b, for details, see Materials and Methods). These three proteins are isoforms of apolipoprotein L (ApoL) 18. As a result, 15 protein spots were isolated and determine by peptide mass fingerprinting and MS/MS analysis. The heel of proteins identified in the NCBI database is shown in table A91. Some proteins haptoglobin, transthyretin (TTR), and apolipoprotein C3 shown in table 1 are present in more than one spot on a gel and have different pI values. Perhaps this phenomenon is due to various post-translational modifications or parti al processing.It was found that only the serum level of ApoA4 was increased (1.8-fold) as compared to the control group (figure 1a). Our findings support other A10reports on change protein levels in serum and cerebrospinal fluid in schizophrenia 19, 20. The decrease in the serum concentration relative to the control group was observed for ApoC3 and for ApoC2 in patients with schizophrenia (figure 1b). This downregulation was on average from 1.8- to 3-fold for ApoC3, and smaller for ApoC2 only 1.25-fold. These proteins are synthesised in the liver and are components of very low-density lipoproteins (VLDLs). Apolipoprotein C2 activates extrahepatic lipoprotein lipase, whereas apolipoprotein C3 can inhibit lipoprotein lipaseFig. 1 Examples of proteins with differential expression in human serum are presented in the enlarged sections of the 2DE profile. a) Apo A4 b) Apo C3 and C2 c) serum amyloid A1 d) transthyretin (the protein is present in two spots because of post-translational mod ifications). Sch schizophrenia.and activate LCAT 21, 22. Previously, it was found that the expression of apolipoproteins is altered in schizophrenia, bipolar disorder, and other psychiatric disorders 23. The authors found that low-density lipoproteins (LDLs) and VLDLs are the most prominent factors differentiating depressed patients from healthy controls, and that plasma unsaturated lipid concentrations are elevated in the depressed group. Thus, there is growing evidence that deregulated lipid homeostasis may play a common role in the pathophysiology of psychiatric disorders such as schizophrenia.Other proteins with a decreased concentration are serum amyloid A1 and TTR (figure 1c and 1d). Inflammatory amyloid A1 is among the supposed acute phase proteins, which have twain direct and indirect bactericidal and/or bacteriostatic properties. According to the classical theory of inflammation, in the acute phase of inflammation, the serum concentration of amyloid A1 increases 100- to 1 000-fold 24, whereas in our study, we observed a 2.3-fold decrease in the serum concentration of this protein in patients with schizophrenia. Perhaps this resultA11 is due to decreased immunity inTable 1. The list of proteins from human serum analysed by 2DE and identified by MALDI-MS/MS after in-gel digestion with trypsin.ProteinNCBIdatabase no.Protein IDpIMW (kDa)Score(individuals/significant)Fold Change(+/-)*Sch vs control1gi1578315962-antitrypsin5.3744.348/45-1.32 0.22gi338305SP 405.7436.717/10-1.31 0.093gi3337390Haptoglobin6.1438.235/29+1.22 0.134gi114318993Transthyretin (dimer)5.1620.2120/84-1.68 0.255gi11957960Apolipoprotein A45.2828.943/32+1.84 0.166gi223976Haptoglobin hp26.2341.739/20-1.38 0.177gi296653Haptoglobin hp26.2341.518/10-1.42 0.118gi296653Haptoglobin hp26.2541.539/15-1.56 0.249gi4507725Transthyretin5.5215.975/52-1.06 0.1310gi4507725Transthyretin5.5215.965/60-1.45 0.2311gi4557323Apolipoprotein C35.2310.8112/70-1.85 0.2112gi4557323Apolipoprotein C35.2310. 8115/68-3.28 0.1613gi4557323Apolipoprotein C25.4211.2104/78-1.25 0.0814gi40316910Serum amyloid A16.2813.5134/90-2.34 0.0915gi19626079Albumin fragment6.2022.456/40-2.06 0.32The fold change is equal to SPi/SPcontrol, where i is the identification number of a spot. Symbols - and + mean a decrease and increase, respectively. Sch schizophrenia.patients with psychiatric disorders or to the presence of comorbidities. In addition, A12it is achievable that a decrease in serum amyloid A1 concentration is related to downregulation of antioxidant-defence enzymes in patients with schizophrenia 6 because high-density lipoproteins (HDLs) inhibit oxidative modification of LDLs via the activity of their associated enzymes and apolipoproteins 25. If HDLs become so-called dysfunctional HDLs because of accumulation of oxidants derived from an inflammatory reaction, such HDLs inhibit the HDL-associated antioxidant enzymes and reduce the ability of apolipoproteins A1 to promote ABCA-1-mediated chole sterol efflux 25. In the literature, there are data on a strong positive relation between cholesterol levels and pathophysiological features of mood disorders. The bring together between mental health (brain) and cholesterol is believed to be based on hypothetical neuron-associated utensils. Cholesterol is an integral component of the plasma membrane of neurons and is present in myelin. Furthermore, cholesterol performs crucial functions in the development, stability, and workings of the synapse 26. Overall, aberrations in cholesterol in a psychiatric illness may substantially affect the mood via synaptic stability and lowered serotonergic activity.In the case of TTR, we observed a decrease in the serum concentration of its dimer and one of monomeric forms (protein 10 in Table 1) among the patients with schizophrenia (1.7-fold and 1.5-fold, respectively), whereas the serum level of another TTR monomeric form was found to be unchanged relative to the control group (protein 9 in Tab le 1).TTR is a liver-derived secretory protein and is the major serum carrier of thyroid hormones thyroxine and tri-iodothyronine. TTR is also gnarly in the transport of retinol via an interaction with retinol-binding proteins. Several studies were conducted in an attempt to identify disease biomarkers that could advance the understanding of the pathogenesis of schizophrenia. In some of these studies, a link between TTR and schizophrenia was found 27, 28. In ref. 28, it was estimated that 3% of TTR in ventricular cerebrospinal fluid A13and 10% of TTR in lumbar cerebrospinal fluid are derived from blood. To assess the involvement of blood TTR in the changes observed in the cerebrospinal fluid of patients with schizophrenia, those authors also studied serum TTR levels in the same people (simultaneously with cerebrospinal fluid collection) by an ELISA. They observed a significant moderate decrease in TTR concentration in serum samples of patients with schizophrenia compared to control s. Nevertheless, there was no familiarity between cerebrospinal-fluid and serum TTR levels in the same individuals, indicating that the protein levels of TTR are regulated by different systems in serum and in cerebrospinal fluid.ConclusionIn the present study, we identified differentially expressed proteins in the serum from patients with schizophrenia by proteomic analysis. We showed differential expression of such proteins as TTR, serum amyloid A1, and apolipoproteins of classes A4 and C3. Furthermore the increase in the expression was found only for apolipoprotein A4, whereas the expression of apolipoprotein C3, TTR, and serum amyloid A1 was decreased.Such alterations of the expression of these proteins may indicate problems with regulation, for example, in the synthesis. On the other hand, the altered protein expression may simply reflect the pathophysiological status of patients with schizophrenia, where these proteins could be candidates for biomarkers. Nevertheless, to confi rm the significance of the altered levels of these proteins in the pathogenesis A14of schizophrenia, and to determine their suitability as biomarkers of schizophrenia, further research is needed.Competing interests. The authors declare that they have no conflicts of interest related to the contents of this article.Funding. This research was made possible in part by a grant from the Russian Science Foundation (14-15-00480, with the exception of the work corresponding to MALDI-TOF MS/MS analysis) and Federal Agency for Scientific Organizations (the part of work corresponding to MALDI-TOF MS/MS analysis).ReferencesKessler RC, Demler O, Frank RG, Olfson M. 2005 Prevalence and treatment of mental disorders 1990 to 2003. N. Engl. J. Med. 352, 2515-2523.J. van Os, S. Kapur Schizophrenia. Lancet (Lond. Engl.), 374 (2009), pp. 635-645H. Moore, A.R. West, A.A. Grace. 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