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Identification of lncRNAs involved in the PCV2 infection of PK-15 cells

ABSTRACT: Porcine circovirus type 2 (PCV2) can cause severe disease in the affected pigs, resulting in massive economic loss for the swine industry. Transcriptomics and proteomics approaches have been widely employed to identify the underlying molecular mechanisms of PCV2 infection. Numerous differentially expressed mRNAs, miRNAs, and proteins, together with their associated signaling pathways, have been identified, paving the way to further validation of their biological functions. Long non-coding RNA (lncRNA) is an important regulator of multiple biological processes. However, rare information regarding its role in the PCV2 infection has been obtained. Hence, in our study, RNA-seq was performed by infecting PK-15 cells with PCV2. Analysis of the differentially expressed genes (DEGs) suggested that cytoskeleton, apoptosis, cell division, and protein phosphorylation were significantly disturbed. Then, using stringent parameters, six lncRNAs were identified. Additionally, the potential targets of the lncRNAs were predicted using both cis- and trans-prediction methods. Interestingly, we found that the HOXB (Homeobox B) gene cluster was probably the target of the lncRNA LOC106505099. Enrichment analysis of the target genes showed that numerous developmental processes were altered during PCV2 infection. Therefore, our study revealed that lncRNAs could affect porcine embryo development through the regulation of the HOXB genes.

ORGANISM(S): Sus scrofa

PROVIDER: GSE149711 | GEO | 2020/06/19

REPOSITORIES: GEO

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Project description:Purpose: Post-weaning multisystemic wasting syndrome (PMWS) is a disease associated with porcine circovirus type 2 (PCV2) infection. One main feature of PMWS is seriously decreased lymphocyte in lymphoid tissues. This research aims to investigate the molecular mechanism of evident lymphocyte depletion in spleen from Yorkshire x Landrace (YL) pigs after infection with PCV2. Methods: At the age of 6 weeks, 15 piglets of Laiwu (LW) and YL pigs were assigned randomly into two groups, respectively: 10 infected pigs and 5 uninfected pigs. Spleen tissues of each group were collected at 35 days post infection with PCV2. Three PCV2-infected YL pigs with apparently pathologic characteristics and mock-infected YL pigs were selected for RNA sequencing, respectively. The RNA extraction, library construction and RNA-Seq were performed on the Illumina HiSeq2500 at Beijing BioMarker Technologies (Beijing, China).Differential expression genes (DEGs) were filtrated from the result of transcriptome. The quantitative real-time PCR was performed to validate the expression patterns of candidate DEGs between LW and YL pigs. Results: The clean reads percolated from the raw reads were mapped to Sus scrofa genome (Sscrofa10.2) using Tophat2 software. The efficiency of alignment was about 76% between the six samples and reference genome, and approximate 50% clean reads were perfectly matched to pig genome. Two samples with high R-squared value (T1-T3: 0.98; T5-T6: 0.97) from PCV2 and mock-infected YL pigs, respectively, was performed differential expression analysis through DESeq after biologic repetition correlation detection. With the criterion of log FC (fold change) > 1.5 and of P value < 0.05, 90 significantly differentially expressed genes were identified between the two groups, of which 57 were up-regulated and 33 were down-regulated. Eleven candidate DEGs were confirmed by quantitative real-time PCR, which expression pattern were generally in accordance with the results of RNA-seq. Conclusions: Many DEGs participate in immune response and some of them are involved in cells proliferation or apoptosis, such as CD81, HGF, CXCL13, KLF11, PSMD4, CYCS and ACTC1. The expression changes of these DEGs may cause the lymphocyte depletion in spleen after challenge with PCV2.

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Identification of lncRNAs involved in the PCV2 infection of PK-15 cells

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