Análise da expressão gênica celular e viral durante a infecção in vitro pelo herpesvírus canino 1
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Data
2017-01-12
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Universidade Estadual Paulista (Unesp)
Resumo
A análise de transcriptoma é essencial para determinar a relação entre as informações codificadas num genoma, a sua expressão e variação genotípica. O transcriptoma é o conjunto completo de transcritos e a quantidade gerada, relacionado a um estágio de desenvolvimento específico ou condição fisiológica da célula. Estes transcritos podem ser mapeados utilizando genomas como referência, para investigação de expressão do genes. Para isso, exige procedimentos de mineração de grandes volumes de dados de RNA-Seq para extrair conhecimentos biológicos. As ferramentas de bioinformática foram desenvolvidas para facilitar e agilizar essas análises, transformando dados em informações. Assim, alguns desses recursos foram empregados na análise de dados gerados pelo RNAseq, a partir de RNAm de cultura celular MDCK infectada por herpesvírus canino tipo 1 (1CaHV-1 - Canid alphaherpesvirus type 1) em diferentes momentos pós infecção. Dessa forma, foi realizado um dual RNAseq, em que foi avaliado tanto a expressão gênica celular do hospedeiro como do patógeno viral, no curso da infecção. Para isso, foram analisados o transcriptoma das atividades celulares e os processos envolvidos no ciclo de infecção viral, até o momento 32h-pi. Assim, foram identificados as atividades de respostas celulares à infecção viral, mecanismos regulatórios induzidos pelo vírus, transcrição de genes virais imediatos, iniciais e tardios. Dentre eles, foi verificado a elevação da expressão do gene COX-2 induzido pela replicação viral, sendo relacionado à hipóxia e apoptose. Tal fato levanta a possibilidade do uso de inibidores seletivos de COX-2 para interrupção do ciclo de replicação viral e no tratamento da infecções por CaHV-1. Também verificou-se que não ocorreu a cascata de indução da expressão de genes do ciclo de infecção forma temporal e ordenada. No qual foi verificado a expressão de genes tardios precocemente, e genes imediatos e iniciais permanecendo a expressão tardiamente. Até o momento foi primeiro trabalho realizado de dual RNAseq para análise de expressão gênica celular e viral para CaHV-1.
Transcriptome analysis is essential to determine the relationship between the information encoded in a genome, its expression and genotypic variation. The transcriptome is the complete set of transcripts and the amount generated, related to a specific developmental stage or physiological condition of the cell. These transcripts can be mapped using genomes as reference for investigation of gene expression. Therefore, were required procedures to mine large volumes of RNA-Seq data to extract biological knowledge. Bioinformatics tools have been developed to facilitate and streamline these analyzes, transforming data into information. Thus, some of these features were employed in the analysis of data generated by RNAseq from canine herpesvirus type 1 (1CaHV-1 - Canid alphaherpesvirus type 1) MDCK cell culture mRNA at different post-infection stages. Thereby, a dual RNAseq was performed, in which both the host cell and the viral pathogen gene expression were evaluated in the course of infection. For this, was analyzed the transcriptome of the cellular activities and the processes involved in the cycle of viral infection, until the moment 32h-pi. Thus, the activities of cellular responses to viral infection, regulatory mechanisms induced by the virus, and transcription of early, early and late viral genes were identified. Among them, was verified the elevation of COX-2 gene expression induced by viral replication, unleashing the hypoxia and apoptosis mechanisms. This fact would suggest the possibility of the use of selective COX-2 inhibitors to interrupt the viral replication cycle and in the treatment of CaHV-1 infections. We found that the induction cascade of gene expression from the infection cycle did not occurred ordered and temporally. The expression of late genes was verified early, and early immediate and early genes remaining expression tardily. To date, it was the first work of dual RNAseq of cellular and viral gene expression analysis for CaHV-1.
Transcriptome analysis is essential to determine the relationship between the information encoded in a genome, its expression and genotypic variation. The transcriptome is the complete set of transcripts and the amount generated, related to a specific developmental stage or physiological condition of the cell. These transcripts can be mapped using genomes as reference for investigation of gene expression. Therefore, were required procedures to mine large volumes of RNA-Seq data to extract biological knowledge. Bioinformatics tools have been developed to facilitate and streamline these analyzes, transforming data into information. Thus, some of these features were employed in the analysis of data generated by RNAseq from canine herpesvirus type 1 (1CaHV-1 - Canid alphaherpesvirus type 1) MDCK cell culture mRNA at different post-infection stages. Thereby, a dual RNAseq was performed, in which both the host cell and the viral pathogen gene expression were evaluated in the course of infection. For this, was analyzed the transcriptome of the cellular activities and the processes involved in the cycle of viral infection, until the moment 32h-pi. Thus, the activities of cellular responses to viral infection, regulatory mechanisms induced by the virus, and transcription of early, early and late viral genes were identified. Among them, was verified the elevation of COX-2 gene expression induced by viral replication, unleashing the hypoxia and apoptosis mechanisms. This fact would suggest the possibility of the use of selective COX-2 inhibitors to interrupt the viral replication cycle and in the treatment of CaHV-1 infections. We found that the induction cascade of gene expression from the infection cycle did not occurred ordered and temporally. The expression of late genes was verified early, and early immediate and early genes remaining expression tardily. To date, it was the first work of dual RNAseq of cellular and viral gene expression analysis for CaHV-1.
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Transcriptoma, RNAseq, Herpesvírus, Infecção, Gene, Cão, Transcriptome, Herpesvirus, Infection, Dog