Genome-Wide mRNA and Long Non-Coding RNA Analysis of Porcine Trophoblast Cells Infected with Porcine Reproductive and Respiratory Syndrome Virus Associated with Reproductive Failure

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is a vertically transmitted reproductive disorder that is typically characterized by miscarriage, premature birth, and stillbirth in pregnant sows after infection. Such characteristics indicate that PRRSV can infect and penetrate the porcine placental barrier to infect fetus piglets. The porcine trophoblast is an important component of the placental barrier, and secretes various hormones, including estrogen and progesterone, to maintain normal pregnancy and embryonic development during pregnancy. It is conceivable that the pathogenic effects of PRRSV infection on porcine trophoblast cells may lead to reproductive failure; however, the underlying detailed mechanism of the interaction between porcine trophoblast (PTR2) cells and PRRSV is unknown. Therefore, we conducted genome-wide mRNA and long non-coding RNA (lncRNA) analysis profiling in PRRSV-infected PTR2. The results showed that 672 mRNAs and 476 lncRNAs were significantly different from the control group after viral infection. Target genes of the co-expression and co-location of differential mRNAs and lncRNAs were enriched by GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis, revealing that most of the pathways were involved in cell nutrient metabolism, cell proliferation, and differentiation. Specifically, the estrogen signaling pathway, the PI3K (PhosphoInositide-3 Kinase)-Akt (serine/threonine kinase) signaling pathway, and the insulin secretion related to embryonic development were selected for analysis. Further research found that PRRSV inhibits the expression of G-protein-coupled estrogen receptor 1 (GPER1), thereby reducing estrogen-induced phosphorylation of AKT and the mammalian target of rapamycin (mTOR). The reduction in the phosphorylation of AKT and mTOR blocks the activation of the GPER1- PI3K-AKT-mTOR signaling pathway, consequently restraining insulin secretion, impacting PTR2 cell proliferation, differentiation, and nutrient metabolism. We also found that PRRSV triggered trophoblast cell apoptosis, interrupting the integrity of the placental villus barrier. Furthermore, the interaction network diagram of lncRNA, regulating GPER1 and apoptosis-related genes, was constructed, providing a reference for enriching the functions of these lncRNA in the future. In summary, this article elucidated the differential expression of mRNA and lncRNA in trophoblast cells infected with PRRSV. This infection could inhibit the PI3K-AKT-mTOR pathway and trigger apoptosis, providing insight into the mechanism of the vertical transmission of PRRSV and the manifestation of reproductive failure.

Integrative transcriptomic profiling of mRNA, miRNA, circRNA, and lncRNA in alveolar macrophages isolated from PRRSV-infected porcine

“…Predominantly employing PAMs as an in vitro infection model, these studies have highlighted that DE-lncRNAs participate in a range of cellular signaling pathways, including viral infection, immune response, NF-kappa B signaling, and toll-like receptor signaling ( 29 , 31 , 33 , 55 ). The transcriptomic analysis of porcine endometrial epithelial cells further revealed the associations of lncRNAs with apoptosis-related genes ( 27 ), and another study on the transcriptome of porcine trophoblast cells suggested PRRSV-induced regulation of apoptosis genes via lncRNAs ( 13 ). These insights provide substantial evidence concerning the mechanisms underlying PRRSV’s vertical transmission and the manifestation of reproductive failure.…”

“…Recent years have witnessed numerous investigations striving to decipher the intricate interplay between lncRNAs and gene expression in the context of PRRSV infection ( 13 , 27 , 29 , 31 , 33 , 55 ). Predominantly employing PAMs as an in vitro infection model, these studies have highlighted that DE-lncRNAs participate in a range of cellular signaling pathways, including viral infection, immune response, NF-kappa B signaling, and toll-like receptor signaling ( 29 , 31 , 33 , 55 ).…”

“…This study discovered that lnc_000397 impedes PRRSV replication by stimulating the expression of interferon-stimulated genes (ISGs) ( 33 ). Moreover, lncRNAs were identified as participants in PRRSV infection by orchestrating the regulation of GPER1 and apoptosis-related genes ( 13 ). Further study extended investigations to lncRNAs, miRNAs, and mRNAs of porcine endometrial epithelial cells post-PRRSV infection, unearthing numerous miRNAs and lncRNAs involved in various pathways such as cell apoptosis and p53 signaling ( 27 ).…”

Fuzhengjiedu San inhibits porcine reproductive and respiratory syndrome virus by activating the PI3K/AKT pathway

“…The expression of NF-κB p65, p-NF-κB and p65 was increased with favipiravir treatment, implying that the antiviral effectivity of favipiravir against peste des petits ruminants virus (PPRV) is mediated by the JAK/STAT and PI3K/AKT pathways [ 23 ]. A recent genome-wide mRNA and long non-coding RNA (lncRNA) search and analysis in PRRSV-infected PTR2 showed that PRRSV infection could inhibit the PI3K/AKT/mTOR pathway and trigger apoptosis [ 24 ]. Another study determined the phosphorylation status of Akt during infection of MARC-145 cells with the highly pathogenic PRRSV strain, HuN4, and provided new evidence of a novel role for the PI3K/Akt pathway in PRRSV infection [ 25 ].…”

“…and analysis in PRRSV-infected PTR2 showed that PRRSV infection could inhibit the PI3K/ AKT/mTOR pathway and trigger apoptosis [24]. Another study determined the phosphorylation status of Akt during infection of MARC-145 cells with the highly pathogenic PRRSV strain, HuN4, and provided new evidence of a novel role for the PI3K/Akt pathway in PRRSV infection [25].…”