Transcriptome Analysis Reveals the Potential Role of Long Noncoding RNAs in Regulating Fowl Adenovirus Serotype 4-Induced Apoptosis in Leghorn Male Hepatocellular Cells

Abstract

Hepatitis-hydropericardium syndrome (HHS) is caused by fowl adenovirus serotype 4 (FAdV-4) and has resulted in considerable economic losses to the poultry industry globally. FAdV-4 elicits apoptosis in host cells. Long noncoding RNAs (lncRNAs) have emerged as important regulatory RNAs with profound effects on various biological processes, including apoptosis. However, it remains unknown whether lncRNAs participate in FAdV-4-induced apoptosis. In this study, RNA sequencing was applied to determine the transcription of cellular lncRNA in leghorn male hepatocellular (LMH) cells infected with FAdV-4. Cellular RNA transcription analysis demonstrated that FAdV-4 infection elicited 1798 significantly differentially expressed (DE) lncRNAs in infected LMH cells at 24 h post-infection (hpi) compared to mock control infection. In addition, 2873 DE mRNAs were also found. Target prediction and analyses revealed that 775 DE lncRNAs whose 671 target mRNAs were among the DE mRNAs were involved in several signaling pathways, including the AMPK signaling pathway, p53 signaling pathway and insulin signaling pathway. From these 775 DE lncRNAs, we identified 71 DE lncRNAs related to apoptosis based on their target gene functions. Subsequently, lncRNA 54128 was selected from the 71 identified DE lncRNAs, and its role in FAdV-4-induced apoptosis was verified. LncRNA 54128 interference significantly suppressed the rate of apoptosis, which was accompanied by reduced BMP4 transcription levels. To the best of our knowledge, this is the first study to analyze host lncRNA transcription during FAdV-4 infection. Our findings provide a better understanding of host responses to FAdV-4 infection and provide new directions for understanding the potential association between lncRNAs and FAdV-4 pathogenesis.

Transcriptome Analysis of Genes Responding to Infection of Leghorn Male Hepatocellular Cells With Fowl Adenovirus Serotype 4

“…Cells were cultured for ~24 h to reach 80% confluence and subsequently infected with FAdV-4 (MOI = 0.1); uninfected LMH cells were used as a mock control. FAdV-4 infected with LMH Cells caused cytopathic effect (CPE), viral titers and viral loads as previously described ( 15 ).…”

Biological features of fowl adenovirus serotype-4

“…It has been reported that viral infection alters the lncRNA transcription profile of infected host cells, which is involved in the establishment and maintenance of persistent infection ( Meng et al., 2017 ). It has been reported that FAdV-4 induces apoptosis in LMH cells by modulating the transcription of the BMP4 gene, which is targeted by lncRNA 54128 ( Wen et al., 2021 ). The miRNAs and lncRNAs whose transcription is induced during FAdV-4 infection, as well as their target mRNAs, are listed in Table 1 .…”

Cellular protein HSC70 promotes fowl adenovirus serotype 4 replication in LMH cells via interacting with viral 100K protein

“…The expression of SUMO-1 activating enzyme subunit ( SAE1 ) was greatly upregulated to restrict FAdV-4 infection in LMH cells ( Hou et al, 2021 ). Additionally, FAdV-4 elicited different expressions of long noncoding RNA ( lncRNA ), microRNA, and mRNA ( Wu et al, 2020 ) in infected LMH cells and lncRNA 54128 was identified to greatly suppress the virus induced apoptosis ( Wen et al, 2021 ). All these results suggested that viral and host factors jointly contributed to the pathogenesis of hypervirulent FAdV-4 strains.…”