Análise de expressão gênica e metilação de dna de pacientes em primeiro episódio psicótico virgens de tratamento

A esquizofrenia é o transtorno mental mais grave e incapacitante dentre os distúrbios psiquiátricos. A demora em instituir tratamento adequado e a duração do primeiro episódio psicótico estão entre os principais fatores de mau prognóstico da doença. O presente projeto propôs a investigação de marcadores genéticos e epigenéticos para o diagnóstico e tratamento da esquizofrenia por meio do estudo da expressão de genes alvos e do estudo do padrão de metilação de DNA (estudo de expressão gênica I). Paralelamente, em células progenitoras de neurônios, estudamos o efeito do silenciamento do gene MBD5 (Methyl-CpG Binding Domain Protein 5), importante para o neurodesenvolvimento e possivelmente relacionado à metilação de DNA (estudo funcional II). Para o estudo I, pacientes em primeiro episódio psicótico (PEP) virgens de tratamento e controles saudáveis foram submetidos à avaliação clínica e à coleta de sangue periférico. Os pacientes ressubmeteram-se às mesmas avaliações após oito semanas do início do tratamento com risperidona, sendo que parte deles também foram reavaliados após um ano de inclusão na pesquisa. Em todas as etapas de seguimento do estudo, o sangue dos pacientes foi recoletado, seguido pela extração de DNA e RNA das amostras. O estudo de expressão gênica I foi realizado por meio da técnica de PCR em tempo real quantitativo, e a análise metilação por meio de sequenciamento após conversão com bissulfito de sódio. Para o estudo funcional II, o silenciamento do gene MBD5 foi realizado em células progenitoras de neurônios. Elas foram submetidas ao sequenciamento de RNA, PCR em tempo real, Western blotting, e à citometria de fluxo. No estudo I os resultados indicaram uma redução na expressão do gene GCH1 (GTP cyclohydrolase 1) e um aumento na expressão dos genes NDEL1 (nudE neurodevelopment protein 1-like 1) e MBP (myelin basic protein) no sangue de pacientes em PEP quando comparados aos controles. Quando comparamos os pacientes antes e após o tratamento, o gene GABRR2 (gamma-aminobutyric acid (GABA) A receptor, rho 2) mostrou-se hipoexpresso após o tratamento com risperidona. Além disso, embora os dados sejam promissores, a expressão de GCH1 não parece ser regulada por metilação de DNA. Quanto ao estudo funcional II, foi observado que o silenciamento do gene MBD5 levava a um desbalanço entre proliferação e diferenciação das células progenitoras de neurônios. Os resultados desse estudo indicaram três genes possivelmente envolvidos na psicose propriamente dita, estando relacionados às vias dopaminérgicas e serotoninnérgicas (GCH1), ao neurodesenvolvimento (NDEL1) e à mielinização (MBP). Além disso, o gene GABRR2, embora não pareça ser alvo direto da risperidona, pode estar associado à resposta ao tratamento ou evolução da doença, indicando, assim, a importância da via GABAérgica. Estes dados poderão futuramente auxiliar a identificação de marcadores biológicos, tanto para a psicose quanto para o tratamento da esquizofrenia, possibilitando uma ação terapêutica precoce e acarretando na redução do tempo de psicose não tratada, o que pode resultar em diminuição da morbidade e melhor qualidade de vida para os pacientes. Também observamos que reduções na expressão de um gene envolvido em doenças do neurodesenvolvimento (MBD5) promovem um estado mais diferenciado que proliferativo de células progenitoras de neurônios, um fenômeno que parece estar presente nas alterações de outros genes envolvidos nessas doenças podendo, assim, ser um mecanismo convergente de doenças do neurodesenvolvimento, como o autismo, a deficiência intelectual e a esquizofrenia.Dados abertos - Sucupira - Teses e dissertações (2013 a 2016

Gene expression changes associated with trajectories of psychopathology in a longitudinal cohort of children and adolescents

We aimed to identify blood gene expression patterns associated to psychopathological trajectories retrieved from a large community, focusing on the emergence and remission of general psychiatric symptoms. Hundred and three individuals from the Brazilian High-Risk Cohort Study (BHRCS) for mental disorders were classified in four groups according to Child Behavior Checklist (CBCL) total score at the baseline (w0) and after 3 years (w1): low–high (L–H) (N = 27), high–low (H–L) (N = 12), high–high (H–H) (N = 34) and low–low (L–L) groups (N = 30). Blood gene expression profile was measured using Illumina HT-12 Beadchips, and paired analyses comparing w0 and w1 were performed for each group. Results: 98 transcripts were differentially expressed comparing w0 and w1 in the L-H, 33 in the H–L, 177 in the H–H and 273 in the L–L. Of these, 66 transcripts were differentially expressed exclusively in the L–H; and 6 only in the H–L. Cross-Lagged Panel Models analyses revealed that RPRD2 gene expression at w1 might be influenced by the CBCL score at w0. Moreover, COX5B, SEC62, and NDUFA2 were validated with another technique and were also differentially regulated in postmortem brain of subjects with mental disorders, indicating that they might be important not only to specific disorders, but also to general psychopathology and symptoms trajectories. Whereas genes related to metabolic pathways seem to be associated with the emergence of psychiatric symptoms, mitochondrial inner membrane genes might be important over the course of normal development. These results suggest that changes in gene expression can be detected in blood in different psychopathological trajectories

Peripheral interleukin-2 level is associated with negative symptoms and cognitive performance in schizophrenia

Although several studies have pointed to a possible role of interleukin 2 (IL-2) in schizophrenia (SZ), association between IL-2 and the different groups of symptoms has not been explored. the objective of this study was to investigate a possible correlation of peripheral IL-2 levels with symptoms and cognitive performance in patients with SZ. in addition, we compared the plasma levels of IL-2 between patients with SZ and healthy controls. Twenty-nine chronically medicated outpatients with SZ according to DSM-IV were compared with twenty-six healthy controls. the patients were evaluated with the Positive and Negative Syndrome Scale (PANSS), the Calgary Depression Scale for Schizophrenia (CDSS), the Clinical Global Impression (CGI) and the Global Assessment of Functioning (GAF). All the participants had blood collected into EDTA tubes by venipuncture between 9:00 and 10:00 AM. Plasma concentrations of IL-2 were determined by cytometric bead array. A computerized neuropsychological battery assessed verbal learning, verbal fluency, working memory, set shifting, executive function, inhibition and intelligence. Patients with SZ had lower levels of IL-2 than healthy controls (p < 0.001). in the SZ group, IL-2 levels were positively correlated with scores in the digit span test (rho = 0.416, P = 0.025) and intelligence (rho = 0.464, P = 0.011). We also found a negative correlation between IL-2 and total score in the negative subscale of PANSS (rho = -0.447, p = 0.015). Our findings suggest that IL-2 may be involved in the mechanisms related to cognitive deterioration and negative symptomatology in schizophrenia. (C) 2014 Elsevier Inc. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Psychiat, Schizophrenia Program PROESQ, BR-04044000 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychiat, Interdisciplinary Lab Clin Neurosci LINC, BR-04039032 São Paulo, BrazilUniv Fed Minas Gerais, Translat Psychoneuroimmunol Grp, BR-31270901 Belo Horizonte, MG, BrazilUniversidade Federal de São Paulo, Dept Psychiat, Schizophrenia Program PROESQ, BR-04044000 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychiat, Interdisciplinary Lab Clin Neurosci LINC, BR-04039032 São Paulo, BrazilWeb of Scienc

Effects of Risperidone on Cytokine Profile in Drug-Naive First-Episode Psychosis

Background: There is robust evidence that schizophrenia is characterized by immune-inflammatory abnormalities, including variations on cytokine levels. the results of previous studies, however, are heterogeneous due to several confounding factors, such as the effects of antipsychotic drugs. Therefore, research on drug-naive first-episode psychosis (FEP) patients is essential to elucidate the role of immune processes in that disorder.Methods: the aim of this study is to compare cytokine levels (IL-2, IL-10, IL-4, IL-6, IFN-gamma, TNF-alpha, and IL-17) in drug-naive FEP patients both before and after treatment with risperidone for 10 weeks, and to investigate possible associations between cytokine levels and clinical responses to treatment and presence of depressive symptoms. It this study, we included 55 drug-naive FEP patients who had repeated measurements of cytokine levels and 57 healthy controls.Results: We found that FEP patients had significantly higher IL-6, IL-10 and TNF-alpha levels than healthy controls. After risperidone treatment, these three cytokines and additionally IL-4 decreased significantly. No significant difference was found between the post-treatment cytokine levels in FEP patients and in healthy controls, suggesting that these alterations in cytokine profiles are a state marker of FEP. No significant association was found between risperidone-induced changes in cytokines and the clinical response to treatment or the presence of depression. There was a significant inverse association between the risperidone-induced changes in IL-10 and the negative symptoms.Conclusions: in conclusion, our results show a specific cytokine profile in FEP patients (monocytic and regulatory T-cell activation) and suggest immunoregulatory effects of risperidone treatment, characterized by suppressant effects on monocytic, Th2, and T-regulatory functions.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao SafraFundacao ABADSJanssenEli LillyLundbeckNovartisRocheUniversidade Federal de São Paulo, Dept Psychiat, São Paulo, BrazilFac Ciencias Med Santa Casa São Paulo, Episode Psychosis Program 1, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morphol & Genet, Div Genet, São Paulo, BrazilDeakin Univ, Dept Psychiat, Geelong, Vic 3217, AustraliaChulalongkorn Univ, Dept Psychiat, Bangkok, ThailandUniversidade Federal de São Paulo, Dept Psychiat, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morphol & Genet, Div Genet, São Paulo, BrazilWeb of Scienc

PRODH Polymorphisms, Cortical Volumes and Thickness in Schizophrenia

Schizophrenia is a neurodevelopmental disorder with high heritability. Several lines of evidence indicate that the PRODH gene may be related to the disorder. Therefore, our study investigates the effects of 12 polymorphisms of PRODH on schizophrenia and its phenotypes. To further evaluate the roles of the associated variants in the disorder, we have conducted magnetic resonance imaging (MRI) scans to assess cortical volumes and thicknesses. A total of 192 patients were evaluated using the Structured Clinical Interview for DSM-IV (SCID), Positive and Negative Syndrome Scale (PANSS), Calgary Depression Scale, Global Assessment of Functioning (GAF) and Clinical Global Impression (CGI) instruments. the study included 179 controls paired by age and gender. the samples were genotyped using the real-time polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP)-PCR and Sanger sequencing methods. A sample of 138 patients and 34 healthy controls underwent MRI scans. One polymorphism was associated with schizophrenia (rs2904552), with the G-allele more frequent in patients than in controls. This polymorphism is likely functional, as predicted by PolyPhen and SIFT, but it was not associated with brain morphology in our study. in summary, we report a functional PRODH variant associated with schizophrenia that may have a neurochemical impact, altering brain function, but is not responsible for the cortical reductions found in the disorder.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo UNIFESP, Disciplina Genet, Dept Morfol & Genet, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, LiNC, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Psiquiatria, São Paulo, BrazilFac Med ABC FMABC, Dept Ginecol & Obstet, Disciplina Genet & Reprod Humana, São Paulo, BrazilFed Univ Para, Lab Genet Humana & Med, BR-66059 Belem, Para, BrazilUniv Fed ABC, Ctr Math Computat & Cognit, Santo Andre, BrazilUniversidade Federal de São Paulo UNIFESP, Disciplina Genet, Dept Morfol & Genet, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, LiNC, São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Psiquiatria, São Paulo, BrazilFAPESP: 2011/50740-5FAPESP: 2007/58736-1Web of Scienc

Obsessive-compulsive symptoms, polygenic risk score, and thalamic development in children from the Brazilian High-Risk Cohort for Mental Conditions (BHRCS)

Background: Thalamic volume measures have been linked to obsessive-compulsive disorder (OCD) in children and adolescents. However, it is unclear if alterations in thalamic volumes occur before or after symptom onset and if there is a relation to the presence of sub-clinical obsessive-compulsive symptoms (OCS). Here, we explore the relationship between OCS and the rate of thalamic volume change in a cohort of children and youth at high risk to develop a mental disorder. A secondary aim was to determine if there is a relationship between OCS and the individual’s OCD polygenic risk score (OCD-PRS) and between the rate of thalamic volume change and the OCD-PRS. Methods: The sample included 378 children enrolled in the longitudinal Brazilian High-Risk Cohort for Mental Conditions. Participants were assessed for OCS and the symmetrized percent change (SPC) of thalamic volume across two time-points separated by 3 years, along with the OCD-PRS. Zero-altered negative binomial models were used to analyze the relationship between OCS and thalamic SPC. Multiple linear regressions were used to examine the relationship between thalamic SPC and OCD-PRS. Results: A significant relationship between OCS and the right thalamus SPC (p = 0.042) was found. There was no significant relationship between changes in thalamic volume SPC and OCD-PRS. Conclusions: The findings suggest that changes in the right thalamic volume over the course of 3 years in children may be associated to OCS. Future studies are needed to confirm these results and further characterize the specific nature of OCS symptoms associated with thalamic volumes

Mapping genomic loci implicates genes and synaptic biology in schizophrenia

Schizophrenia has a heritability of 60-80%1, much of which is attributable to common risk alleles. Here, in a two-stage genome-wide association study of up to 76,755 individuals with schizophrenia and 243,649 control individuals, we report common variant associations at 287 distinct genomic loci. Associations were concentrated in genes that are expressed in excitatory and inhibitory neurons of the central nervous system, but not in other tissues or cell types. Using fine-mapping and functional genomic data, we identify 120 genes (106 protein-coding) that are likely to underpin associations at some of these loci, including 16 genes with credible causal non-synonymous or untranslated region variation. We also implicate fundamental processes related to neuronal function, including synaptic organization, differentiation and transmission. Fine-mapped candidates were enriched for genes associated with rare disruptive coding variants in people with schizophrenia, including the glutamate receptor subunit GRIN2A and transcription factor SP4, and were also enriched for genes implicated by such variants in neurodevelopmental disorders. We identify biological processes relevant to schizophrenia pathophysiology; show convergence of common and rare variant associations in schizophrenia and neurodevelopmental disorders; and provide a resource of prioritized genes and variants to advance mechanistic studies

Mapping genomic loci prioritises genes and implicates synaptic biology in schizophrenia

Schizophrenia has a heritability of 60–80%1, much of which is attributable to common risk alleles. Here, in a two-stage genome-wide association study of up to 76,755 individuals with schizophrenia and 243,649 control individuals, we report common variant associations at 287 distinct genomic loci. Associations were concentrated in genes that are expressed in excitatory and inhibitory neurons of the central nervous system, but not in other tissues or cell types. Using fine-mapping and functional genomic data, we identify 120 genes (106 protein-coding) that are likely to underpin associations at some of these loci, including 16 genes with credible causal non-synonymous or untranslated region variation. We also implicate fundamental processes related to neuronal function, including synaptic organization, differentiation and transmission. Fine-mapped candidates were enriched for genes associated with rare disruptive coding variants in people with schizophrenia, including the glutamate receptor subunit GRIN2A and transcription factor SP4, and were also enriched for genes implicated by such variants in neurodevelopmental disorders. We identify biological processes relevant to schizophrenia pathophysiology; show convergence of common and rare variant associations in schizophrenia and neurodevelopmental disorders; and provide a resource of prioritized genes and variants to advance mechanistic studies

Schizophrenia and 22q11 deletion syndrome: Characterization of relevant genes

Introdução: A esquizofrenia é o transtorno mental mais grave e incapacitante entre os distúrbios psiquiátricos. Ela é uma doença complexa e com fenótipo heterogêneo. Dentre os fatores genéticos que parecem ter um papel na etiologia da esquizofrenia está a deleção 22q11.2. Objetivos: Investigar alterações cromossômicas, polimorfismos dos genes UFD1L e ZDHHC8, mutações no gene TBX1 e variações no número de cópias na esquizofrenia e na síndrome da deleção 22q11.2, e correlacionar com achados de avaliações genético-clínicas, psiquiátricas, neuropsicológicas e de neuroimagem. Métodos: Um total de 200 portadores de esquizofrenia, 200 indivíduos controles e 10 portadores do fenótipo clínico da síndrome da deleção 22q11.2, mas sem a deleção, participaram do presente estudo. Os pacientes com esquizofrenia foram estudados por citogenética clássica e Multiplex Ligation-dependent probe amplification. Os polimorfismos rs5992403 (gene UFD1L) e rs175174 (gene ZDHHC8) foram investigados em pacientes com esquizofrenia e controles por meio de PCR em tempo real com sonda TaqMan. Outros polimorfismos do gene UFD1L foram analisados, rs5746744 e rs1547931, por Restriction Fragment Length Polymorphism. Mutações no gene TBX1 foram investigadas em portadores do fenótipo clínico da síndrome da deleção 22q11.2, mas sem a deleção, por meio de sequenciamento genômico. As variações no número de cópias foram analisadas por meio da metodologia de array em pools. Os pacientes com esquizofrenia também foram avaliados por testes neuropsicológicos e por neuroimagem estrutural. Resultados: Todos os cariótipos estudados foram normais. Foi encontrada um paciente com a deleção de 1,5 megabases na região 22q11.2. Os polimorfismos rs5992403 (UFD1L) e rs175174 (ZDHHC8) foram associados com a idade de acometimento da esquizofrenia. Além disso, todos os polimorfismos investigados parecem desempenhar um papel na morfologia cerebral e em habilidades cognitvas. Nenhuma mutação foi encontrada no gene TBX1, apenas polimorfismos, em portadores do fenótipo clínico da 22q11DS. Foram encontradas três regiões amplificadas em pools de DNAs de portadores de esquizofrenia: 1p36.32, 2q37.3 e 22q11.21. Conclusões: O estudo permitiu avaliar a participação de fatores genéticos em determinadas características da esquizofrenia, propiciando um melhor entendimento sobre a etiologia e fisiopatologia dessa doença complexa.Background: Schizophrenia is a severe, persistent, debilitating and poorly understood psychiatric disorder. It is a complex disease with heterogeneous fenotype. Among the genetic factors that might have a role in schizophrenia, it is included 22q11.2 deletion. Objectives: We aimed to investigate chromosomal abnormalities, UFD1L and ZDHHC8 polymorphisms, TBX1 mutations and copy number variations in schizophrenia and 22q11.2 deletion syndrome and associate them with clinical genetics, psychiatric, neuropsychological and neuroimaging data. Methodology: A total of 200 schizophrenia patients, 200 healthy controls and 10 patients who have the 22q11.2 syndrome phenotype but no detectable deletion were selected. Schizophrenia patients were investigated through classical karyotyping by G-banding and Multiplex Ligationdependent probe amplification. UFD1L rs5992403 and ZDHHC8 rs175174 polymorphisms were genotyped in schizophrenia patients and healthy controls by Real Time PCR using TaqMan. UFD1L rs5746744 and rs1547931 polymorphisms were genotyped by Restriction Fragment Length Polymorphism. Tbx1 mutations were investigated in patients who have the 22q11.2 syndrome phenotype but no detectable deletion by sequencing. Affymetrix 6.0 microarrays in a pool of DNA samples was used to detect copy number variations. Schizophrenia patients were evaluated by neuropsychological tests and structural neuroimaging. Results: All karyotypes were normal. One patient presented a 1.5 megabases deletion in 22q11.2 region. UFD1L rs5992403 and ZDHHC8 rs175174 polymorphisms were associated with age at onset of schizophrenia. Moreover, all studied polymorphisms may have a role in brain morphology and cognition. No mutation, only polymorphisms, in TBX1 gene was found in 22q11.2 patients. Three regions were amplified in DNA pools of schizophrenia patients: 1p36.32, 2q37.3 e 22q11.21. Conclusion: This study evaluated the role of genetic factors in some schizophrenia fenotypes, providing a better understanding of its etiology and pathophysiology.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 2008/56464-7TEDEBV UNIFESP: Teses e dissertaçõe