Navegando por Palavras-chave "Epigenetics"

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    Butirato, produto da microbiota intestinal, previne dano aos podócitos via mecanismos epigenéticos e dependentes de receptores acoplados a proteína G
    (Universidade Federal de São Paulo (UNIFESP), 2017-07-31) Felizardo, Raphael Jose Ferreira [UNIFESP]; Camara, Niels Olsen Saraiva [UNIFESP]; http://lattes.cnpq.br/8098379714093877; http://lattes.cnpq.br/9834749309169873; Universidade Federal de São Paulo (UNIFESP)
    Introduction: Chronic Kidney Disease (CKD) is characterized by structural abnormalities and progressive decrease of kidney function until complete loss of filtration capacity. Damages to podocytes, specialized cells involved on filtration process, are one of the major causes that lead to a CKD. The literature comprises a large number of experimental attempts that focus on mitigating the damage to the podocytes. In recent years, many studies point to the gut microbiota as a modulator of intestinal and extraintestinal diseases through the generation of short chain fatty acids (SCFA). It is already known that chronic kidney patients have an imbalanced gut microbiota and lower production of SCFA. Thus, we have explored the role of butyrate, an AGCC able to regulate epigenetic processes and activate G protein coupled receptors (GPR), during the progression of experimental nephropathy induced by adriamycin, focusing mainly on the protection of podocytes. Methods: Wild type mice in a Balb/c background, Gpr109a-/- mice and Gpr109a-/-.Gpr43-/- mice were induced to develop glomerulopathy by a single dose of adriamycin and treated with butyrate. Results: Wild type mice treated with butyrate showed improvement of renal function, associated to a preserved podocyte layer in the glomerular basement membrane and reduction of pro-inflammatory and pro-fibrotic markers in the kidneys. Particularly, butyrate modulated the activity of enzymes involved on epigenetic modifications in the kidneys and changed the frequency of histone markers (H3K9Ac, H3K4me3 and H3K9me3) in the promoter region of the genes encoding synaptopodin, podocin and NEPH-1 in the podocytes. Concomitantly, treatment with butyrate was not sufficient to improve the renal function of Gpr109a -/- mice and Gpr109a-/-.Gpr43-/-mice. Activation of the receptors was associated with the regulations of small GTPases activity Rac1 and Cdc42 and maintenance of the organization of actin filaments in the podocytes grown in vitro. Conclusion: Our results demonstrate that butyrate exerts important effects on podocyte homeostasis during experimental nephropathy through epigenetic and GPR109a receptors-mediated mechanisms.
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    Desregulação Emocional Grave Em Crianças E Adolescentes Com Tdah: Avaliação Multidimensional Dos Fatores Ambientais, Neuropsicológicos E Epigenéticos
    (Universidade Federal de São Paulo (UNIFESP), 2017-08-02) Schuch, Viviane [UNIFESP]; Muszkat, Mauro [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Severe Emotional Dysregulation (SED) is defined as dysfunction in controlling the physiological excitement of the brain caused by strong emotions and is often associated with the difficulties of inhibiting inappropriate behavior in response to positive or negative emotions. Although emotional dysregulation is not part of the cardinal symptoms of Attention Deficit Hyperactivity Disorder (ADHD) according to the DSM-5 (Diagnostic and Statistical Manual of Mental Disorders 5th Edition), characterized by the symptomatological triad inattention, hyperactivity and impulsivity, SED is often associated with a more unfavorable evolution in the presence of comorbidities and neuropsychological dysfunctions, which result in problems in social, educational and family environments. The aim of this thesis was to perform a multidimensional neuropsychological assessment of environmental and epigenetic factors in children aged 6 to 15 years with diagnosis of ADHD and symptoms of SED. The presence of SED was defined as the sum of the Child Behavior Checklist Anxiety / Depression, Aggression and Attention subscales (AAA) subscales was greater than 210. Two studies were conducted. In Study I, we evaluated the neuropsychological performance of 52 children with ADHD, 26 of them with SED (experimental group) compared to 26 children without SED (control group). All cases underwent interdisciplinary neuropsychological assessment at the NANI - UNIFESP ADHD clinic, including intellectual level determination, Concentrated and selective attention evaluation and ecological executive functions through the BRIEF Behavior Rating Inventory of Executive Functions (BRIEF) and CBCL questionnaires. Survey of environmental factors through a “Structured questionnaire of socio-environmental factors” (QEFSA). Considering the established confidence level (p≤ 0.05), there was a difference between the groups, with lower performance of the SED group in all subscales of the CBCL, with the exception of Retreatment / Depression and Somatic Complaints. There was a statistically significant difference between the presence of SED and the degree of executive dysfunction in BRIEF. The presence of SED was associated with a higher hyperactivity and impulsivity score, according to DSM-5 criteria. There was a negative correlation between the SED and age at onset of the ADHD symptoms, and the SED was more frequent in the ADHD of earlier onset. Regarding the environmental profile, the SED was associated with exposure to previous traumatic experiences reported in the QEFSA. These findings suggest that SED symptoms may be markers of evolution and expression of the neuropsychological profile of ADHD and if identified early can help us to identify children in situations of vulnerability. In study II the objective was to recognize the differences in the methylation profile of the genome of children and adolescents with ADHD (15 with SED and 14 without SED). The genomic methylation profile of the 40 children was analyzed using the 450k (Illumina) technology. Principal component analysis shows that the gender difference represents the greatest source of variability in the data. We identified some probes that are differentially methylated or variously methylated, identifying candidate genes to be epigenetic markers of DEG. The first most differentially methylated probe between the two groups is in the ADK gene, related posynaptic plasticity, altered dopaminergic function, attentional impairment, and deficits in cognitive domains and sleep regulation. The second most differentially methylated probe is in the KCNIP4 gene, implicated in synaptic plasticity and neuronal proliferation and differentiation systems. In the Gene Ontology (GO) analysis, many of the major differentially methylated genes are related to the nervous system, providing clues as to how the epigenetic modulation of the phenotype of emotional self-regulation occurs.
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    DNA methylation profile of the DKK2 gene as a biomarker in patients with colorectal cancer
    (Funpec-Editora, 2017) Silva, T. D. [UNIFESP]; Felipe, A. V. [UNIFESP]; Vidigal, V. M. [UNIFESP]; Saad, S. S. [UNIFESP]; Forones, N. M. [UNIFESP]
    Purpose: Epigenetic changes can be detected in precancerous lesions, suggesting that may be involved in the early stages of carcinogenesis. The methylation of specifics genes has been correlated with the outcome of many different types of cancers. This study compared the levels of DNA methylation between normal and tumor tissues from patients with colorectal cancer (CRC). Methods: Candidate genes were screened using the MethylProfiler T PCR Array System and the Dickkopf-related protein 2 (DKK2) genes were selected for study. DNA methylation of these genes was assessed by polymerase chain reaction-high-resolution melting (PCR-HRM) analysis. Results: Out of the 112 patients studied, 68 were controls with a mean age (SD)of 59.9 years (13.4) and 45 were patients with CRC with a mean age (SD) of 64.6 years (13.6). Among the patients with CRC, 25 were women, 28 were diagnosed with colon cancer and 18 were diagnosed with rectal cancer. The number of patients with Stage I, II, III, and IV of the disease, as per the TNM classification, were 2 (4.3%), 20 (43.4%), 18 (39.1%), and 2 (4.3%), respectively. Furthermore, the DKK2 gene had a higher methylation profile in the CRC tissues when compared to normal mucosa (p < 0.001), whereas the methylation profile analysis was not statistically significant when comparing the stages of the disease (p = 0.078). Conclusion: The presence of differentially methylated sequences of the DKK2 gene between the groups showed that the methylation changes of these genes could potentially be prognostic and predictive markers in CRC.
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    Efeito da suplementação do fruto da palmeira juçara (Euterpe edulis Mart.) nas modificações epigenéticas e estado inflamatório em monócitos de adultos obesos
    (Universidade Federal de São Paulo, 2020-06-23) Santamarina, Aline Boveto [UNIFESP]; Pisani, Luciana Pellegrini [UNIFESP]; Oyama, Lila Missae [UNIFESP]; http://lattes.cnpq.br/7125541171554727; http://lattes.cnpq.br/3983527783636073; http://lattes.cnpq.br/7371478593039938; Universidade Federal de São Paulo (UNIFESP)
    Introdução: A obesidade já está estabelecida como problema de saúde pública e intrinsecamente relacionada ao quadro de inflamação subclínica crônica, bem como alterações epigenéticas. Com isso houve crescimento na busca por alimentos capazes de atuar no combate e prevenção da obesidade. Assim, o fruto da palmeira juçara possui potencial inexplorado devido à sua composição rica em antocianinas, ácidos graxos insaturados e fibras alimentares. Objetivo: Avaliar o efeito do consumo da polpa liofilizada do fruto da palmeira juçara (Euterpe edulis Mart.), sobre os parâmetros inflamatórios, epigenéticos e metabólicos relacionados à obesidade. Métodos: 27 ratos Wistar machos foram divididos em 4 grupos experimentais: dieta controle; dieta hiperlipídica; dieta hiperlipídica + juçara 0,25%; e dieta hiperlipídica + juçara 0,5% por 7 dias. Foi avaliado o consumo alimentar, massa corporal e de tecidos adiposos viscerais, hipotálamo e fígado. No soro foram avaliadas enzimas hepáticas, e frações lipídicas circulantes por ensaios colorimétricos; o acúmulo ectópico de lipídios no fígado foi avaliado por análise histológica. Avaliou-se a expressão proteica por western blotting da via TLR4 / NFκB e produção de citocinas (IL-10, IL-6, TNF-α) por ELISA. Em modelo clínico, 27 adultos com obesidade (IMC 30,0 – 39,9 kg/m2), foram randomizados em 2 grupos, em uso ou não (placebo) de suplementação de juçara por 6 semanas. Foi realizada avaliação antropométrica e da ingestão alimentar, perfil de ácidos graxos presentes no soro; foi avaliada a via TLR4 / NFκB, por expressão gênica (RT-qPCR) e proteica (western blotting) além da produção de citocinas por ELISA em monócitos isolados do sangue. Em células THP-1, foram analisados os efeitos do ácido graxo oleico e da cianidina 3-rutinosideo isoladamente ou em associação sobre a expressão gênica (RT-qPCR) e proteica (western blotting e multiplex) de marcadores inflamatórios relacionas à via TLR4 / NFκB. Resultados: A dose de juçara de 0,25% preveniu ganho de peso corporal, e reduziu inflamação nos tecidos central e periféricos bem como lesão hepática mediante à dieta hiperlipídica em modelo animal. Em indivíduos obesos a suplementação melhorou o perfil de ácidos graxos do soro, perfil inflamatório e produção de citocinas além de modular o perfil epigenético. Os achados do estudo desenvolvido in vitro demonstrou que a combinação de ácido oleico e cianidina 3-rutinosideo teve melhores efeitos anti-inflamatórios em monócitos e macrófagos por modulação de NFκB e PPARγ. Conclusão: O consumo da juçara pode ser considerado um potencial nutracêutico, já que o efeito sinérgico entre o ácido oleico e antocianinas, pode potencializar efeitos anti-inflamatórios observados possibilitando do estabelecimento futuro de estratégias para o tratamento e prevenção da obesidade e suas comorbidades.
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    Efeitos da dieta hiperlipídica sobre a expressão de receptores de leptina e sua ação sobre modificações epigenéticas em células mesenquimais multipotentes extraídas do tecido adiposo de camundongos.
    (Universidade Federal de São Paulo, 2022-12-20) Bispo, Andressa França de Sousa [UNIFESP]; Vale, Maria Isabel Cardoso Alonso [UNIFESP]; Correa, Lucia Maria Armelin [UNIFESP]; http://lattes.cnpq.br/4266175652800598
    Uma dieta hiperlipídica leva à obesidade, uma condição que gera importantes alterações no tecido adiposo branco (TAB). As células tronco mesenquimais derivadas do tecido adiposo (Adipose derivedStromalCells – ASCs), são responsáveis por gerar novos adipócitos, expandindo o tecido adiposo. Alterações na expressão de fatores de transcrição e de marcas epigenéticas no núcleo de ASCs podem ter efeitos na proliferação e na adipogênese dessas células afetando a expansãosaudável (hiperplásica) do TAB versus expansão patológica (hipertrófica) do TAB em indivíduos obesos. A leptina é uma adipocina com efeitos no sistema nervoso central, que incluem a indução do gasto energético e da saciedade. Também, tem grande relevância na ativação de células imunes e na modulação da resposta inflamatória. Sabe-se que, na obesidade, os efeitos centrais da leptina são prejudicados, pois os aumentos no acúmulo de TAB e nos níveis de leptina são acompanhados por resistência hipotalâmica à leptina, assim, seus efeitos locais podem prevalecer e contribuir para a expansão do TAB e aumento da inflamação. Esse fenômeno tem sido considerado resistência à leptina e pode ser resultado de deficiências em vários níveis na via de sinalização da leptina, incluindo acesso reduzido do hormônio ao seu receptor devido a alterações na expressão do receptor e/ ou alterações na transdução de sinal pós-receptor. O TAB é um ambiente rico em leptina, mas as ações locais da leptina sobre a adipogênese não foram completamente investigadas. Assim, se faz necessário avaliar as contribuições da sinalização parácrina da leptina e de seus receptores na obesidade, em células mesenquimais multipotentes do TAB, objetivo do presente projeto. Considerando que as ASCs são células progenitoras que residem em um nicho altamente enriquecido em leptina (secretadas por adipócitos) no TAB de obesos, hipotetizamos que a obesidade (via ação da leptina em ASCs), levaria a importantes alterações nas enzimas promotoras de marcas epigenéticas, que culmina na queda da expressão de alguns genes comumente silenciados na obesidade, especialmente aqueles relacionados à adipogênese. Para estudarmos estas alterações avaliamos a expressão do receptor de leptina por PCR em Tempo Real, em ASCs isoladas do TAB subcutâneo e epididimal de animais controles e obesos (induzidos por dieta hiperlipídica), a fim de correlacionar os dados com demais resultados do grupo. Adicionalmente, tratamos ASCs de obesos com leptina a fim de entender seu efeito parácrino (in vitro) sobre estas mesmas marcas. Nossos resultados mostraram que os animais obesos (induzidos pela dieta hiperlipídica DHL) apresentaram um aumento de ~30% na massa corpórea, maior glicemia de jejum (~50%) e intolerância à glicose. A expressão de ambos os receptores Lepr1 e Lepr2 foram diminuídos em ~40 e 45%, respectivamente, em ASCs de obesos em comparação aos animais controle. A leptina (como esperado) não foi expressa nestas células, tal qual LepR3. Detectamos uma diminuição na expressão da H3K27 acetilada nas ASCs de obesos, e da Acly, que parece ter um importante papel na acetilação da histona 3. Essa marca epigenética contribui para a ativação da transcrição de genes ligados a adipogênese. Adicionalmente, a expressão da Acly e da enzima demetilase Kdm6b (enzima que remove grupos metil para posterior ação de acetilases) diminuiu quando ASCs foram tratadas in vitro com leptina. Assim, sugerimos que a sinalização aumentada de leptina em ASCs diminui a expressão de ATP Citrato Liase, levando à redução de substrato para acetilação de H3K27 o que pode contribuir para o silenciamento de genes comumente observados na obesidade.
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    Estresse oxidativo, comportamento e modificações epigenética em modelo animal de hiper-homocisteinemia
    (Universidade Federal de São Paulo (UNIFESP), 2014) Rezende, Marina Mastelaro de [UNIFESP]; D’Almeida, Vânia [UNIFESP]
    Objetivo: Analisar como o aumento plasmático de homocisteína por diferentes fontes e durações influencia as funções cognitivas, considerando as alterações bioquímicas e epigenéticas decorrentes desse acúmulo em camundongos. Metodologia: Um total de 125 camundongos da linhagem C57Black foi separado em quatro grupos experimentais e seus tratamentos foram mantidos por três diferentes durações, totalizando 12 grupos. Os tratamentos consistiram na adição de L-metionina nas concentrações de 0,5 e 1% (grupos M05 e M1) e DL-homocisteína a 0,09% (grupo H) na água de beber dos animais. Esse procedimento foi mantido por 2, 4 e 6 meses, a fim de se analisar não apenas as respostas aos tratamentos, mas também a influência de sua duração. Em todos os períodos houve também o acompanhamento de animais que não tiveram a água de beber suplementada, como controles (grupo CT). Separados por 1 semana a cada teste, antes do término do final dos tratamentos, os animais foram submetidos aos testes comportamentais: Caixa de atividade, Reconhecimento de objetos novos, Labirinto aquático de Morris e Campo aberto, a fim de se averiguar possíveis alterações cognitivas devidas aos tratamentos. Após os tratamentos e avaliações comportamentais, os animais foram eutanasiados por decapitação e tiveram sangue, cérebro, gordura, coração, fígado e fêmur coletados para os testes relacionados ao balanço redox, epigenética e estado geral dos animais. Os dados são apresentados como média ± desvio padrão. Resultados e conclusões: Os tratamentos foram capazes de aumentar os níveis plasmáticos de homocisteína em relação aos animais CT, porém o perfil de respostas variou conforme tratamento e duração. As quantidades de cisteína plasmática também tiveram um padrão dependente de tratamento e duração. Com estes dados, pode-se concluir que as respostas metabólicas decorrentes da indução de hiper-homocisteinemia são abrangentes e específicas para cada situação. Considerando os marcadores de estresse oxidativo eritrocitários (catalase e superóxido dismutase), aparentemente os tratamentos com metionina têm consequências mais importantes após 2 meses de suplementação, enquanto com homocisteína, 6 meses. Levando em consideração as quantidades de glutationa plasmática e eritrocitária, não parece ter havido influência devida ao tratamento, o que pode ser devido ao diferenciado metabolismo de tióis em roedores. Não houve também alteração dos parâmetros biométricos nos grupos experimentais, o que pode ser devido à magnitude dos aumentos alcançados. Poucas foram as alterações observadas nos testes comportamentais, e elas foram mais devidas a alterações decorrentes do envelhecimento dos animais do que devidas aos diferentes períodos de tratamento. As alterações bioquímicas centrais foram também mais relacionadas ao envelhecimento do que ao tratamento. Pode-se concluir que a indução de hiper-homocisteinemia por diferentes formas de tratamento resulta em modificações bioquímicas e comportamentais específicas ao tipo e duração de cada tratamento.
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    Identificação do perfil de hidroximetilação do DNA genômico durante a progressão do melanoma
    (Universidade Federal de São Paulo (UNIFESP), 2020-05-18) Leite, Guilherme Burgarelli [UNIFESP]; Jasiulionis, Miriam Galvonas [UNIFESP]; Universidade Federal de São Paulo
    The loss of 5hmC is considered a hallmark in many types of cancer. Different studies in melanoma showed a profound decrease in hydroxymethylation in melanomas in comparison with healthy melanocytes. However, these studies are limited to two very specific steps of the melanoma progression. Using a murine model established in our lab consisting of cell lines corresponding to distinct stages of the melanoma progression, this study aims to assess the DNA hydroxymethylation dynamics during this process, and the impact of the tumor microenvironment in DNA methylation and hydroxymethylation in melanoma cells. Here we show that Tet2-silenced cell lines, cell lines isolated from tumor in situ and lung metastasis present characteristics compatible with increased aggressiveness and a build-up of 5mC and 5hmC modifications in genes which regulate ion channels and transporters and signaling pathways regulators. Furthermore, we identified a 5hmC signature in specific promoters and gene body suggesting a safeguarding role of this epigenetic mark against melanoma progression. Finally, 9 in 10 genes identified as differentially expressed in the 4C11+ metastatic cell line in comparison with the melan-a, 4C and 4C11- cell lines in our murine melanoma progression model showed correlation with poor overall survival in melanoma patients (COL4A1, DOCK8, IRF8, HLA-A [mouse H2-K1 analogous], PAK1, PARM1, PDE3A, RUNX1 e TNC). Seven of these genes also presented the described 5hmC epigenetic signature (Dock8, Irf8, H2-K1 [HLA-A analogous], Pak1, Parm1, Pde3a e Tnc). Together, our results show the importance of epigenetic profiling in different stages of the melanoma progression as a prognostic and predictive marker and also as a therapy target, and the role of the tumor microenvironment in these modifications.
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    Identificação e análise funcional de genes desregulados por metilação envolvidos em etapas iniciais e tardias da progressão do melanom
    (Universidade Federal de São Paulo (UNIFESP), 2020-07-30) Papaiz, Debora Dangelo [UNIFESP]; Jasiulionis, Miriam Galvonas [UNIFESP]; Universidade Federal de São Paulo
    DNA mutations and epigenetic alterations are responsible for the malignant transformation and progression of tumors. Different patterns of methylation in cancer cells are capable of silencing tumor suppressor genes; increasing the expression of genes previously silenced; and of supporting chromosomal instability and loss of cellular imprinting. Melanoma is a very aggressive skin cancer due to its high probability of developing metastasis, and its incidence in population has increased more than any other cancer. Our laboratory established a model of malignant transformation from murine non-tumorigenic melanocytes through cycles of adhesion blockage, resulting in cell lines in different stages of the melanoma progression. Through the analysis of DNA methylation profile (ERRBS) and gene expression (RNA-seq) of each cell line, it was possible to evaluate how DNA methylation regulates gene expression along melanoma progression. Furthermore, we studied genes that had different methylation patterns in promoter and gene body regions and negative or positive correlation with gene expression, respectively. This study provided the identification of potentially important genes for the process of malignant transformation of melanocytes and melanoma metastasis, as well as identified possibly new prognostic biomarkers for melanoma patients.
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    In vitro use of latency reversing agents to overcome HIV latency
    (Universidade Federal de São Paulo (UNIFESP), 2017-02-23) Samer, Sadia [UNIFESP]; Diaz, Ricardo Sobhie [UNIFESP]; Miyuki, Telma Oshiro; http://lattes.cnpq.br/1101949023118413; http://lattes.cnpq.br/0846508761438062; http://lattes.cnpq.br/8843157636360084; Universidade Federal de São Paulo (UNIFESP)
    The use of antiretroviral therapy (ART) dramatically suppresses HIV replication to undetectable levels and can prolong the life of an infected individual, but low level viremia and proviral DNA are detected in majority of patients on ART even with intensifications of antiretroviral regimens. Furthermore virus typically rebounds within one to eight weeks once the treatment is interrupted. This rebound of viral load is mainly due to quiescent, integrated, replication competent latent proviral pool. Being latent, it remains hidden from the immune cells and the infection persists for years until and unless the cells carrying it die by apoptosis or are succumbed to cytopathic effect, once viral replication is induced by antigenic stimuli. Reducing this latent pool may help the patients to reach at least a transient drug-free remission of their disease. Latent HIV proviruses are silenced as a result of deacetylation and methylation of histones located at the long terminal repeats (LTRs) and lack of some viral (Tat) and host factors (NFKB, NFAT, AP1, PTEFb) that assist viral transcription. Breakthrough of latency reversal agents (LRAs) has helped to break this dormant HIV state so that HIV protein expression can be induced, followed by the clearance of that cell by viral cytopathic effects, immune cells among individuals under suppressive ART. Since HIV transcription is the interplay between host and viral factors, it utterly needs active cell state with upregulated transcription factors and their access to viral genome. We used two different approaches to activate the proviral compartment. One at chromatin level, Using Phytohaemagglutinin and histone deacetylase and methyltransferse inhibitors (HDACi and MTi, respectively) and the other approach was co-stimulation of the cells using Thalidomide coupled with Phytohaemagglutinin as TCR-agonist. Both ways were found efficient to reactivate/purge the virus from the patients on long term ART and undetectable viral load but the nature of the purged virus was different in both cases. The samples treated with HDACi/MTis were to be used later to sensitize monocyte derived dendritic cells (MDDCs) as MDDCs vaccine. So, we needed a high titer of virus. Viral purging was observed in samples of antiretroviral treated patients at different time points of supernatant collection, but the viral load showed decline over time in samples treated with HDACI/MTis (Nicotinamide and Chaetocin/BIX01294 respectively) in autologous culture. When cultures were shifted to an allogenic system to avoid host restriction factors, and to use healthy CD4 receptors to make new infection in order to obtain better viral yield, the virus behaved the same way, showing a viral load decline over time towards extinction. So the required viral titer could not be obtained due to massive misfit viral pool in the culture. Samples treated with Thalidomide showed an abrupt viral purging in 6/7 samples after 48 hours treatments with PHA and Thalidomide, in contrast to HDACi/MTis, where the average purging time was 5 days (Min 3, Max 7) and 6.5 days (Min 3, Max 29) respectively. Because the treatment with thalidomide was just for a mini in vitro test to check its latency reversal potential, we did not expand the virus from this treatment. We visually inspected the viral particles by electron microscopy, purged from both treatments. In first case where the samples were treated with HDACi/MTIs, we exclusively found virus with small size, diffused and unclear matrix and in most of cases lack of double membrane. In the second case where samples were treated with thalidomide, virus particles were of normal virus size, clear matrix and double membrane. We therefore hypothesize that long term suppressive ART will reduce the number of replicating competent HIV over time, since those strains will randomly interrupt latency and be eliminated by ART. This phenomenon might lead to the accumulation of defective HIV in proviral compartment. Finding of this cryptic defective provirus pool in autologous/allogenic system and defects seen by electron microscopy from the samples treated with HDACs/MTis and on the other hand normal viral population seen in the samples treated with a co-stimulator are directing to design new strategies aimed at eliminating cells harboring this defective proviral pool in order to further decrease the latent HIV reservoir among individuals under long term suppressive ART.
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    Investigação da herança intergeracional - via espermatozoides - de alterações na expressão de miRNAs relacionados a transtornos psiquiátricos
    (Universidade Federal de São Paulo (UNIFESP), 2020-09-30) Campagnoli, Gabriella Mesas [UNIFESP]; Teixeira, Taiza Stumpp [UNIFESP]; Universidade Federal de São Paulo
    Introduction: Epigenetics is identified as an important component of the inheritance of several conditions, among which “missing heritability”, such as schizophrenia, since the genetic aspects of this disorder do not explain its heritability. Besides, the fact that the environment is crucial for the manifestation and development of schizophrenia brings to light the importance of the involvement of epigenetic mechanisms in this disorder. As schizophrenia is considered a developmental disorder, or the use of experimental models that allow investigating the development of this disorder from the early life is essential,especially with regard to respect for its molecular aspects. MiRNAs play an important role in neurogenesis, involved in behavioral changes and psychiatric disorders. Thus, an epigenetic analysis of brain regions and sperm can contribute to the understanding of epigenetic aspects involved in schizophrenia and specific about their epigenetic inheritance. Aims: To investigate the profile of miRNAs related to schizophrenia in sperm, prefrontal cortex and hippocampus of SHR rats. Methods: Sperm miRNAs, prefrontal cortex and hippocampus were extracted, quantified and reproduced in the construction of libraries for further sequencing. The captured data were used for bioinformatics and statistical analyzes, as well as an interactive analysis was carried out between differently expressed miRNAs and target genes through the Metascape and Cytoscape platforms. Results: We observed differential expression of miRNAs in the three types of tissue/cell. A bioinformatics analysis has shown that the vast majority of differently expressed miRNA target genes, both for sperm and the prefrontal cortex and hippocampus, are using the nervous system and its important pathways to relate to behavior, neurodevelopment and synaptic processes. Among the genes included, the miRNAs differently expressed, 12 coincide with the genes expressly expressed in patients with schizophrenia, or that reinforce SHR as a good model for the study of these disorders. Conclusion: the male rats of the SHR lineage show alterations in the expression of miRNAs related to neurological involvement and schizophrenia in sperm, prefrontal cortex and hippocampus, as the SHR model is useful for epigenetic studies and other molecular aspects related to schizophrenia. Finally, this study suggests that the SHR model may be useful for future studies on the epigenetic inheritance of miRNAs.
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    Methyl-CpG-Binding Protein (MBD) Family: Epigenomic Read-Outs Functions and Roles in Tumorigenesis and Psychiatric Diseases
    (Wiley-Blackwell, 2016) Gigek, Carolina Oliveira [UNIFESP]; Chen, Elizabeth Suchi [UNIFESP]; Smith, Marilia Arruda Cardoso [UNIFESP]
    Epigenetics is the study of the heritable changes on gene expression that are responsible for the regulation of development and that have an impact on several diseases. However, it is of equal importance to understand how epigenetic machinery works. DNA methylation is the most studied epigenetic mark and is generally associated with the regulation of gene expression through the repression of promoter activity and by affecting genome stability. Therefore, the ability of the cell to interpret correct methylation marks and/or the correct interpretation of methylation plays a role in many diseases. The major family of proteins that bind methylated DNA is the methyl-CpG binding domain proteins, or the MBDs. Here, we discuss the structure that makes these proteins a family, the main functions and interactions of all protein family members and their role in human disease such as psychiatric disorders and cancer. (C) 2015 Wiley Periodicals, Inc.
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    Modulação dos fatores epigenéticos nos momentos iniciais da infecção pelo HIV-1 em células T CD4+ in vitro
    (Universidade Federal de São Paulo (UNIFESP), 2017-03-31) Nunes, Jorge Meneses [UNIFESP]; Janini, Luiz Mario Ramos [UNIFESP]; http://lattes.cnpq.br/5713863164263481; http://lattes.cnpq.br/6876504227479535; Universidade Federal de São Paulo (UNIFESP)
    Current research trends in human immunodeficiency virus 1/acquired immune deficiency syndrome (HIV-1/AIDS) and epigenetic phenomena have been focusing on elucidating the effect of epigenetics on the viral genome and the subsequent effects on the viral life cycle, particularly during latency. However, the relationship between HIV-1 and possible epigenetic modifications of the host cell genome has not been thoroughly studied. In this context, the objective of the present study was to identify epigenetic factors that are regulated at the beginning of HIV-1 infection in activated and quiescent CD4+ T cells. We analyzed the gene and protein expression of a specific set of epigenetic targets, global DNA methylation, and HIV-1 replication kinetics for 36 hours after infecting CD4+ T cells in vitro. The cells were obtained from the blood bags of eight healthy donors. Among the 84 epigenetic targets analyzed, infection affected the expression of the genes aurora kinase B (AURKB), aurora kinase C (AURKC), and DNA methyltransferase 3B (DNMT3B). The global DNA methylation percentage exhibited opposing trends in the infected and uninfected (control) groups, increasing with time in the former and decreasing in the latter. Moreover, the detected levels of HIV-1 were higher in activated than in quiescent cells. Thus, the epigenetic targets AURKB, AURKC, and DNMT3B and the global DNA methylation profile are regulated during HIV-1 replication in CD4+ T cells, and this regulation can be influenced by the activation state of the cell at the time of infection.
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    Mudanças no padrão de metilação do DNA na progressão do melanoma
    (Universidade Federal de São Paulo (UNIFESP), 2020-07-30) Rius, Flavia Eichemberger [UNIFESP]; Jasiulionis, Miriam Galvonas [UNIFESP]; Universidade Federal de São Paulo
    Melanoma is the cancer derived from melanocytes, cells located in the basal layer the epidermis which are responsible for melanin production. In Brazil there we 6,260 deaths by melanoma at 2018, with this being the skin cancer with highe number of deaths. In this work, a model constituted by cell lineages correspondent different stages of melanoma progression was used to disclose DNA methylati marks which could characterize each stage. The experiment called ERRB (Enhanced Reduced Representation Bisulfite Sequencing) was realized to access methyl cytosines in CpG context in the cell lines, at base pair level. Methylati states were analyzed in malignant transformation of melanocytes as well as acquisition of metastatic capability of melanoma. In melanoma maligna transformation it was observed biggest hypomethylation in genic promoters, whilst melanoma metastatization it was observed significant hypermethylation in tho regions. The analysis of regulatory capability of those genic promoters showed th hypomethylated promoters have an important relationship to regulation of ge expression for ransformation signature as well as for metastatization of melanom signature. Genes which present hypomethylated promoters in maligna transformation signature as well as in metastatization signature enrich for pathwa related to embryonic development, specially of return to epigenetic a transcriptional state of neural crest cells. Hypermethylated promoters we associated to cell membrane regulation in the signature of melanom metastatization. Using TCGA data, multivariate analyses of ten of the gen identified in this study show correlation between the level of methylation and surviv of patients with melanoma for four genes, namely NEFM, PLXNB1, SEMA7A a SYT5. Analysis of the methylome of cell lineages composing the model of study h provided a great amount of data, which could still be explored to identify not on new biomarkers for prognostic, but also to disclose mechanisms involved malignant transformation of melanocytes as well as in melanoma progression.
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    Physical exercise as an epigenetic modulator of brain plasticity and cognition
    (Pergamon-Elsevier Science Ltd, 2017) Fernandes, Jansen [UNIFESP]; Arida, Ricardo Mario [UNIFESP]; Gomez-Pinilla, Fernando
    A large amount of evidence has demonstrated the power of exercise to support cognitive function, the effects of which can last for considerable time. An emerging line of scientific evidence indicates that the effects of exercise are longer lasting than previously thought up to the point to affect future generations. The action of exercise on epigenetic regulation of gene expression seem central to building an "epigenetic memory" to influence long-term brain function and behavior. In this review article, we discuss new developments in the epigenetic field connecting exercise with changes in cognitive function, including DNA methylation, histone modifications and microRNAs (miRNAs). The understanding of how exercise promotes long-term cognitive effects is crucial for directing the power of exercise to reduce the burden of neurological and psychiatric disorders.