Identificação de biomarcadores na fibrogênese pulmonar da paracoccidioidomicose
Carregando...
Arquivos
Data
2018-02-27
Autores
Santos, Amanda Ribeiro
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Paracoccidioidomicose (PCM) é uma micose sistêmica causada por fungos do gênero Paracoccidioides; suas principais formas clinicas são aguda/subaguda, crônica (FC) e residual. A maioria dos pacientes com a FC da doença, mesmo após tratamento eficaz, apresenta sequelas, principalmente fibrose pulmonar (FP) e enfisema. Os problemas sociais, econômicos e psicológicos desencadeados pela FP são subestimados. Apesar da fibrogênese na PCM ser reconhecida como um processo precoce, seus mecanismos não são totalmente conhecidos. No presente estudo, determinamos o perfil proteômico sérico de pacientes com PCM-FC com diferentes desfechos de FP. Vinte e nove pacientes com PCM pulmonar foram acompanhados periodicamente e as amostras de soro sanguíneo foram coletadas em quatro momentos: antes do tratamento (M0), cura clínica (M1), cura sorológica (M2) e cura aparente (M3). O grupo controle foi constituído por amostras de quinze indivíduos fumantes, pareados com idade e sexo dos pacientes avaliados. Ao final do tratamento (M3), os pacientes foram submetidos à avaliação de tomografia computadorizada e dois grupos de pacientes foram formados com base nos resultados de PF: PF menor (n = 17) e PF maior (n = 12). As proteínas de elevada abundância no soro foram removidas e, em seguida, submetidas à análise proteômica shotgun através do sistema nanoACQUITY UPLC-Xevo QTof MS. A identificação e quantificação das proteínas foram realizadas usando o software PLGS Expression E. Análise de enriquecimento e análise de rede de interação proteína-proteína (PPI) foram realizadas usando o software String. Antes do tratamento, os pacientes com PF menor, em comparação aos controles, apresentavam 91 proteínas diferencialmente expressas (60 sub-reguladas 31 superreguladas). Soro de pacientes com maior PF apresentaram 70 proteínas diferencialmente expressas (66 sub-reguladas 4 reguladas). A maioria dessas proteínas estavam envolvidas na regulação do processo celular e resposta inflamatória. Nas condições ensaiadas, o estudo permitiu identificar as proteínas APOH, A2M, GC e SERPINA1 como alvos envolvidos nos mecanismos moleculares da fibrogênse pulmonar na paracoccidioidomicose. Foi observado, ainda, que o momento de cura clínica apresentou maior atividade pró-fibrótica durante o seguimento dos pacientes. Além disso, foram identificadas 20 proteínas candidatas a biomarcadores no prognóstico da fibrose pulmonar, conforme segue: família das hemoglobulinas (HBB, HBD, HBG2, HBE1, HBG1, HBA1), ENO3, A2M, APOH, GC, IGHG-1, -2 e 3, IGLC1, -2 e -3, HP, SERPINA1, LRG1 e PZP. Nossos achados fornecem novos alvos para estudos futuros envolvendo a FP na PCM, assim como biomarcadores séricos em potencial.
Paracoccidioidomycosis (PCM) is systemic mycosis caused by fungi of the genus Paracoccidioides; its main clinical forms are acute / subacute, chronic (CF) and residual. Most FC-PCM patients, even after effective treatment, present sequelae, mainly pulmonary fibrosis (PF) and emphysema. The social, economic and psychological problems triggered by PF are underestimated. Although fibrogenesis in PCM is recognized as an early process, its mechanisms are not fully known. In the present study, we aimed to determine the serum proteomic profile of patients with FC-PCM with different (PF) outcomes. Twenty-nine patients with pulmonary PCM patients were followed up periodically, and blood serum samples were collected in four moments: before treatment (M0), clinical cure (M1), serological cure (M2), and apparent cure (M3). For control group (n=15), serum samples of sex and age-matched smokers were also collected. At the end of treatment (M3), the patients were submitted to CT Scan evaluation and two groups of patients were formed based on PF outcomes: minor PF (n=17) and major PF (n=12). High-abundance proteins were removed from serum and submitted to shotgun proteomic assay using a nanoACQUITY UPLC-Xevo QTof MS system. The identification and quantification of proteins were performed using PLGS Expression E software. Enrichment analysis and protein-protein interaction (PPI) network analysis were performed using String software. Before treatment, patients with minor PF, in comparison to controls, showed 91 differentially expressed proteins (60 down-regulated and 31 up-regulated). Serum of patients with major PF exhibited 70 differentially expressed proteins (66 down-regulated and 4 up-regulated). Most of these proteins are involved in the regulation of cellular process and inflammatory response. The results allowed the identification of APOH, A2M, GC and SERPINA1 as targets involved in the molecular mechanisms of pulmonary fibrogenesis in paracoccidioidomycosis. It was also observed that the clinical cure presented greater pro-fibrotic activity during the follow-up of the patients. In addition, 20 candidate biomarkers were identified in the prognosis of pulmonary fibrosis, as follows: hemoglobin family (HBB, HBD, HBG2, HBE1, HBG1, HBA1), ENO3, A2M, APOH, GC, IGHG-1, -2 and 3, IGLC1, -2 and -3, HP, SERPINA1, LRG1 and PZP. Our findings provide new targets for further studies involving mechanisms of PF in PCM, as well as for promising potential biomarkers.
Paracoccidioidomycosis (PCM) is systemic mycosis caused by fungi of the genus Paracoccidioides; its main clinical forms are acute / subacute, chronic (CF) and residual. Most FC-PCM patients, even after effective treatment, present sequelae, mainly pulmonary fibrosis (PF) and emphysema. The social, economic and psychological problems triggered by PF are underestimated. Although fibrogenesis in PCM is recognized as an early process, its mechanisms are not fully known. In the present study, we aimed to determine the serum proteomic profile of patients with FC-PCM with different (PF) outcomes. Twenty-nine patients with pulmonary PCM patients were followed up periodically, and blood serum samples were collected in four moments: before treatment (M0), clinical cure (M1), serological cure (M2), and apparent cure (M3). For control group (n=15), serum samples of sex and age-matched smokers were also collected. At the end of treatment (M3), the patients were submitted to CT Scan evaluation and two groups of patients were formed based on PF outcomes: minor PF (n=17) and major PF (n=12). High-abundance proteins were removed from serum and submitted to shotgun proteomic assay using a nanoACQUITY UPLC-Xevo QTof MS system. The identification and quantification of proteins were performed using PLGS Expression E software. Enrichment analysis and protein-protein interaction (PPI) network analysis were performed using String software. Before treatment, patients with minor PF, in comparison to controls, showed 91 differentially expressed proteins (60 down-regulated and 31 up-regulated). Serum of patients with major PF exhibited 70 differentially expressed proteins (66 down-regulated and 4 up-regulated). Most of these proteins are involved in the regulation of cellular process and inflammatory response. The results allowed the identification of APOH, A2M, GC and SERPINA1 as targets involved in the molecular mechanisms of pulmonary fibrogenesis in paracoccidioidomycosis. It was also observed that the clinical cure presented greater pro-fibrotic activity during the follow-up of the patients. In addition, 20 candidate biomarkers were identified in the prognosis of pulmonary fibrosis, as follows: hemoglobin family (HBB, HBD, HBG2, HBE1, HBG1, HBA1), ENO3, A2M, APOH, GC, IGHG-1, -2 and 3, IGLC1, -2 and -3, HP, SERPINA1, LRG1 and PZP. Our findings provide new targets for further studies involving mechanisms of PF in PCM, as well as for promising potential biomarkers.
Descrição
Palavras-chave
fibrose pulmonar, proteômica, paracoccidioides, paracoccidioidomicose, marcadores bioquímicos