Expressão de microRNAs Circulantes em Doença de Crohn e Retocolite Ulcerativa
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Data
2020-03-02
Autores
De Síbia, Carina de Fátima [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Introdução: As doenças inflamatórias intestinais são doenças crônicas representadas principalmente pela Doença de Crohn (DC) e Retocolite Ulcerativa (RCU). A DC caracteriza-se por focos de inflamação transmural que podem afetar qualquer segmento do trato gastrointestinal (TGI).Pode ser classificada em tipo inflamatório, estenosante ou penetrante/fistulizante. Na RCU, a doença é caracteristicamente restrita à superfície da mucosa e submucosa. Os mecanismos envolvidos na patogênese de ambas as doenças ainda são desconhecidos. Ressaltamos a importância de mecanismos de regulação gênica, em particular os microRNAs (miRNAs) que estão envolvidos em processos biológicos importantes, e têm sido evidenciados como biomarcadores com potencial utilidade clínica no diagnóstico, determinação do prognóstico e tratamento dos pacientes. Objetivo: Identificar miRNAs circulantes com expressão significativamente alterada no plasma de pacientes com DC ou RCU, comparando os pacientes respondedores vs. não respondedores ao tratamento. Metodologias: Foram coletadas amostras de plasma de pacientes atendidos no Ambulatório de Doenças Inflamatórias Intestinais do Hospital das Clínicas da Faculdade de Medicina de Botucatu e diagnosticados com DC (n=25) ou RCU (n=30). A DC foi classificada de acordo com a classificação de Montreal e a atividade da doença foi baseada no índice CDAI. A RCU foi classificada de acordo com a extensão da doença em proctite, hemicolite esquerda e pancolite e a atividade da doença foi classificada de acordo com o escore de Mayo. O plasma foi utilizado para extração do RNA e quantificação da expressão de miRNAs na plataforma TaqMan Low Density Array. Para análises de dados utilizamos o programa Expression Suite com cut-off de nível de alteração, foldchange (FC) ≥ 2,0 e p<0,05. Adicionalmente, utilizamos ferramentas de bioinformática (mirDIP, miRTarBase e ToppGeneSuite) para interpretação do significado biológico dos dados. Resultados: A idade média dos pacientes com DC foi de 44,4 anos e 60% eram do sexo feminino. Em relação às características clínicas, 64% dos pacientes foram diagnosticados com a doença entre 17-40 anos de idade, 36% apresentam a localização ileo colonica, 40% com comportamento estenosante, 4% apresentavam-se em atividade clínica da doença e 44% estavam em uso de terapia biológica. Identificamos 37 miRNAs significativamente alterados (FC≥2 e p≤0,05) comparados aos indivíduos saudáveis. Em pacientes respondedores ao tratamento, 22 miRNAs estavam com expressão diminuída e 3 com expressão aumentada. Nos pacientes não respondedores, 10 miRNAs estavam com expressão diminuída e 2 com expressão aumentada. Nos pacientes com RCU, a idade média foi de 51,3 anos e 53% eram do sexo feminino, 70% dos pacientes tiveram comprometimento de todo o cólon, 27% apresentavam atividade moderada e 33% dos pacientes estavam em uso de aminossalicílicos. Identificamos 37 miRNAs alterados, dos quais 18 miRNAs estavam com expressão diminuída e 1 miRNA com expressão aumentada em pacientes respondedores ao tratamento. Nos pacientes não respondedores, foram identificados 5 miRNAs com expressão diminuída e 3 com expressão aumentada. Conclusões: Diferentes miRNAs podem constituir biomarcadores preditivos nas diferentes formas de DII: DC e RCU.
Introduction: Inflammatory bowel diseases (IBD) are chronic diseases mainly represented by Crohn´s disease (CD) and Ulcerative Colitis (UC). CD is characterized by transmural inflammation foci that can affect any segment of the gastrointestinal tract (TGI. It can be classified as inflammatory, stenosing or penetrating/fistulizing. In UC, the disease is restricted to the surface of the mucosa and submucosa. The mechanisms involved in the pathogenesis of both diseases are still unknown. We emphasize the importance of mechanisms of gene regulation, in particular microRNAs (miRNAs) that are involved in important biological processes and have been shown to be biomarkers with potential clinical utility in the diagnosis, determination of prognosis and treatment of patients. Objectives: Identify circulating miRNAs with significantly altered expression in the plasma of patients with CD or UC, comparing responders vs. responders. non-responders to treatment. Methods: Plasma samples were collected from patients seen at the Intestinal Inflammatory Diseases Outpatient Clinic from Botucatu Medical School (UNESP) diagnosed with CD (n = 25) or UC (n = 30). CD was classified according to the Montreal classification and disease activity was based on the CDAI index. The UC was classified according to the extent of the disease into proctitis, left hemicolitis and pancolitis and the activity of the disease was classified according to the Mayo score. Plasma was used to extract RNA and quantify the expression of miRNAs on the TaqMan Low Density Array platform. For data analysis we used the Expression Suite program with cut-off level of change, foldchange (FC) ≥ 2.0 and p <0.05. Additionally, we use bioinformatics tools (mirDIP, miRTarBase and ToppGene Suite) to interpret the biological meaning of the data. Results: The average age of patients with CD was 44.4 years and 60% were female. Regarding the clinical characteristics, 64% of the patients were diagnosed with the disease between 17-40 years of age, 36% had the ileocolonic localization, 40% with stenosing behavior, 4% were in clinical activity of the disease and 44% were using biological therapy. We identified 37 significantly altered miRNAs (FC≥2 and p≤0.05) compared to healthy individuals. In patients who responded to treatment, 22 miRNAs were with decreased expression and 3 with increased expression. In non-responders, 10 miRNAs were with decreased expression and 2 with increased expression. In patients with UC, the mean age was 51.3 years and 53% were female, 70% of the patients had involvement of the entire colon, 27% had moderate activity and 33% of the patients were using aminosalicylics. We identified 37 altered miRNAs, of which 18 miRNAs were with decreased expression and 1 miRNA with increased expression in patients who responded to treatment. In non-responders, 5 miRNAs with decreased expression and 3 with increased expression were identified. Conclusions: Distinct sets of miRNAs can may represent predictive biomarkers in thedifferent forms of IBD: CD and UC.be
Introduction: Inflammatory bowel diseases (IBD) are chronic diseases mainly represented by Crohn´s disease (CD) and Ulcerative Colitis (UC). CD is characterized by transmural inflammation foci that can affect any segment of the gastrointestinal tract (TGI. It can be classified as inflammatory, stenosing or penetrating/fistulizing. In UC, the disease is restricted to the surface of the mucosa and submucosa. The mechanisms involved in the pathogenesis of both diseases are still unknown. We emphasize the importance of mechanisms of gene regulation, in particular microRNAs (miRNAs) that are involved in important biological processes and have been shown to be biomarkers with potential clinical utility in the diagnosis, determination of prognosis and treatment of patients. Objectives: Identify circulating miRNAs with significantly altered expression in the plasma of patients with CD or UC, comparing responders vs. responders. non-responders to treatment. Methods: Plasma samples were collected from patients seen at the Intestinal Inflammatory Diseases Outpatient Clinic from Botucatu Medical School (UNESP) diagnosed with CD (n = 25) or UC (n = 30). CD was classified according to the Montreal classification and disease activity was based on the CDAI index. The UC was classified according to the extent of the disease into proctitis, left hemicolitis and pancolitis and the activity of the disease was classified according to the Mayo score. Plasma was used to extract RNA and quantify the expression of miRNAs on the TaqMan Low Density Array platform. For data analysis we used the Expression Suite program with cut-off level of change, foldchange (FC) ≥ 2.0 and p <0.05. Additionally, we use bioinformatics tools (mirDIP, miRTarBase and ToppGene Suite) to interpret the biological meaning of the data. Results: The average age of patients with CD was 44.4 years and 60% were female. Regarding the clinical characteristics, 64% of the patients were diagnosed with the disease between 17-40 years of age, 36% had the ileocolonic localization, 40% with stenosing behavior, 4% were in clinical activity of the disease and 44% were using biological therapy. We identified 37 significantly altered miRNAs (FC≥2 and p≤0.05) compared to healthy individuals. In patients who responded to treatment, 22 miRNAs were with decreased expression and 3 with increased expression. In non-responders, 10 miRNAs were with decreased expression and 2 with increased expression. In patients with UC, the mean age was 51.3 years and 53% were female, 70% of the patients had involvement of the entire colon, 27% had moderate activity and 33% of the patients were using aminosalicylics. We identified 37 altered miRNAs, of which 18 miRNAs were with decreased expression and 1 miRNA with increased expression in patients who responded to treatment. In non-responders, 5 miRNAs with decreased expression and 3 with increased expression were identified. Conclusions: Distinct sets of miRNAs can may represent predictive biomarkers in thedifferent forms of IBD: CD and UC.be
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Palavras-chave
MicroRNAs, Doença de Crohn, Retocolite Ulcerativa, Doença Inflamatória Intestinal, Crohn´s Disease, Ulcerative Colitis