Staphylococcus aureus Avoids Autophagy Clearance of Bovine Mammary Epithelial Cells by Impairing Lysosomal Function

Jing

Oxidative Medicine and Cellular Longevity

et al. 2021

Mastitis is a common disease in the dairy industry that causes huge economic losses worldwide. Exosomes (carrying proteins, miRNA, lncRNA, etc.) play a vital role in the regulation of immune response. lncRNA can play a variety of regulatory roles by combining with protein, RNA, and DNA. The expression of mRNA and lncRNA in exosomes derived from bovine mammary epithelial cells infected by S. aureus is rarely understood. To explore this issue, RNA sequencing analysis was performed on exosomes derived from S. aureus-infected and noninfected MAC-T cells. Analysis of the sequencing results showed that there were 186 differentially expressed genes, 431 differentially expressed mRNAs and 19 differentially expressed lncRNAs in the exosomes derived from S. aureus-infected and noninfected MAC-T cells. By predicting lncRNA target genes, it was found that 19 differentially expressed lncRNAs all acted on multiple mRNAs in cis and trans. GO analysis revealed that differentially expressed genes and lncRNA target genes played significant roles in such metabolism (reactive oxygen species metabolic processes), transmembrane transport, cellular response to DNA damage stimulus, and response to cytokines. KEGG enrichment indicated that lncRNA target genes gathered in the TNF pathway, Notch pathway, MAPK pathway, NF-kappa B pathway, Hippo pathway, p53 pathway, reactive oxygen species metabolic processes, and longevity regulating pathway. In summary, all data indicated that differentially expressed gene, mRNA, and lncRNA in transcriptional profiling of exosomes participated in bacterial invasion and adhesion, oxidative stress, inflammation, and apoptosis-related signaling pathway. The data obtained in this study would provide valuable resource for understanding the lncRNA information in exosomes derived from dairy cow mammary epithelial cells and conduced to the study of S. aureus infection in dairy cow mammary glands.

“…Mastitis is an inflammation of the udders of dairy cows that is almost always caused by pathogenic microorganisms [ 21 ]. It seriously harms dairy farming and the dairy industry.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional Profiling of Exosomes Derived from Staphylococcus aureus‐Infected Bovine Mammary Epithelial Cell Line MAC‐T by RNA‐Seq Analysis

Jing

Oxidative Medicine and Cellular Longevity

et al. 2021

“…However, although autophagy has an antibacterial effect, there is evidence that pathogens have developed several ways to evade this mechanism (70). S. aureus can avoid autophagy clearance in bMECs by impairing lysosome function (30). Which may be an important mechanism of S. aureus to evade the host's immune response.…”

Section: Discussionmentioning

confidence: 99%

“…The model of intracellular infection was established in our previous study (30). In brief, MAC-T cells were seeded on 6-well cell plates, and when cells were cultured to monolayers, S. aureus (OD 600nm = 0.8-1.2) was inoculated with (treatment group) or without (control group) an MOI of 8:1.…”

Section: Intracellular Infection Modelmentioning

confidence: 99%

See 1 more Smart Citation

RNA-Seq Whole Transcriptome Analysis of Bovine Mammary Epithelial Cells in Response to Intracellular Staphylococcus aureus

Wang

Geng

et al. 2020

Front. Vet. Sci.

Self Cite

Staphylococcus aureus (S. aureus), a common mastitis pathogen widespread in the natural environment of dairy farms, is capable of invading mammary epithelial cells making treatment difficult. However, the mechanism of the response of bovine mammary epithelial cell to S. aureus invasion remains elusive. In this study, transcriptomic analysis and bioinformatics tools were applied to explore the differentially expressed RNAs in bovine mammary epithelial cells (bMECs) between the control and S. aureus-treated group. A total of 259 differentially expressed mRNAs (DEmRNAs), 27 differentially expressed microRNAs (DEmiRNAs), and 21 differentially expressed long non-coding RNAs (DElncRNAs) were found. These RNAs mainly enrich the inflammatory response, immune response, endocytosis, and cytokine-cytokine receptor interaction. qRT-PCR was used to analyze the quality of the RNA-seq results. In particular, to the defense mechanism of bovine mammary epithelial cells against intracellular S. aureus, the PPAR signaling pathway and the genes (ACOX2, CROT, and NUDT12) were found to be up-regulated to promote the production of peroxisomes and ROS, DRAM1 expression was also up-regulated to facilitate the activation of autophagy, indicating that the above mechanisms were involved in the elimination of intracellular S. aureus in bovine mammary epithelial cells.

“…Mastitis is an in ammation of the udders of dairy cows that is almost always caused by pathogenic microorganisms [21]. It seriously harms dairy farming and the dairy industry.…”

Section: Discussionmentioning

confidence: 99%

LncRNA Profiling of Exosomes Derived From Staphylococcus Aureus-induced Bovine Mammary Epithelial Cell Lines MAC-T

Yu¹,

Jing²,

Chen³

et al. 2020

Preprint

Background: Exosomes (carrying proteins, miRNA, lncRNA, etc.) play a vital role in participating in the occurrence and development of mastitis. LncRNA can play a variety of regulatory roles by combining with protein, RNA, and DNA. The expression of mRNA and lncRNA in exosomes derived from bovine mammary epithelial cells infected by S. aureus is rarely understood.Methods: We used ultracentrifugation to separate exosomes derived from S. aureus- stimulated MAC-T cells. After extracting exosomal RNA, RNA sequencing was performed. We screened differentially expressed genes, mRNA and lncRNA, and predicted their functions through KEGG and GO analysis. 6 mRNA and lncRNA were randomly selected for RT-qPCR verification.Results: Analysis of the sequencing results showed that there were 186 differentially expressed genes, 431 differentially expressed mRNAs and 19 differentially expressed lncRNAs in the exosomes derived from S. aureus-infected and non-infected MAC-T acells. By predicting lncRNA target genes, it was found that 19 differentially expressed lncRNAs all acted on multiple mRNAs in cis and trans. GO analysis revealed that differentially expressed genes and lncRNA target genes played significant roles in such metabolism, transmembrane transport, cellular response to DNA damage stimulus, response to cytokines. KEGG enrichment indicated that lncRNA target genes gathered in the TNF pathway, Notch pathway, phosphatidylinositol signaling system and p53 pathway. Conclusion: The data suggested that S. aureus induced changes in genes, mRNA, and lncRNA in exosomes secreted by MAC-T cells, which furtherly triggered other cells damage through exosomal delivery. This study would provide valuable resource for understanding the lncRNA information in exosomes derived from dairy cow mammary epithelial cells, and conduced to the study of S. aureus infection in dairy cow mammary glands.