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Home / Equine Research Bank / PLoS One / Characterization of basal and lipopolysaccharide-induced mic...
PloS one2017; 12(5); e0177664; doi: 10.1371/journal.pone.0177664

Characterization of basal and lipopolysaccharide-induced microRNA expression in equine peripheral blood mononuclear cells using Next-Generation Sequencing.

Abstract: The innate immune response to lipopolysaccharide contributes substantially to the morbidity and mortality of gram-negative sepsis. Horses and humans share an exquisite sensitivity to lipopolysaccharide and thus the horse may provide valuable comparative insights into this aspect of the inflammatory response. MicroRNAs, small non-coding RNA molecules acting as post-transcriptional regulators of gene expression, have key roles in toll-like receptor signaling regulation but have not been studied in this context in horses. The central hypothesis of this study was that lipopolysaccharide induces differential microRNA expression in equine peripheral blood mononuclear cells in a manner comparable to humans. Illumina Next Generation Sequencing was used to characterize the basal microRNA transcriptome in isolated peripheral blood mononuclear cells from healthy adult horses, and to evaluate LPS-induced changes in microRNA expression in cells cultured for up to four hours. Selected expression changes were validated using quantitative reverse-transcriptase PCR. Only miR-155 was significantly upregulated by LPS, changing in parallel with supernatant tumor necrosis factor-α concentration. Eight additional microRNAs, including miR-146a and miR-146b, showed significant expression change with time in culture without a clear LPS effect. Target predictions indicated a number of potential immunity-associated targets for miR-155 in the horse, including SOCS1, TAB2 and elements of the PI3K signaling pathway, suggesting that it is likely to influence the acute inflammatory response to LPS. Gene alignment showed extensive conservation of the miR-155 precursor gene and associated promoter regions between horses and humans. The basal and LPS-stimulated microRNA expression pattern characterized here were similar to those described in human leukocytes. As well as providing a resource for further research into the roles of microRNAs in immune responses in horses, this will facilitate inter-species comparative study of the role of microRNAs in the inflammatory cascade during endotoxemia and sepsis.
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Publication Date: 2017-05-26 PubMed ID: 28552958PubMed Central: PMC5446123DOI: 10.1371/journal.pone.0177664Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research study investigates how the lipopolysaccharide type of bacteria influences the inflammation response through an exploration of microRNA activity in horses’ blood cells. It uses advanced sequencing technology to map changes in microRNA expression. The findings corroborate with human leukocytes’ responses, providing a valuable resource for understanding sepsis and inflammatory reactions.

Objective of the Research

  • The primary aim of this research was to discern the impact of lipopolysaccharide (LPS), a component often found in gram-negative bacteria, on the microRNA expression in equine peripheral blood mononuclear cells (PBMCs). This objective stems from a broader interest in understanding the innate immune response to LPS, which is considered a significant contributor to the morbidity and mortality associated with gram-negative sepsis.

Research Methodology

  • The researchers utilized Next-Generation Sequencing technology from Illumina to analyze the microRNA transcriptome in the PBMCs isolated from healthy horses.
  • These blood cells were cultured for up to four hours, and then LPS-induced changes in microRNA expression were evaluated.
  • The researchers validated certain expression changes using quantitative reverse-transcriptase PCR.

Key Findings

  • The study identified significant upregulation of only miR-155 microRNA in response to LPS. This expression change paralleled with the increased concentration of tumor necrosis factor-α, a protein responsible for regulating immune cells, in the cell culture’s supernatant.
  • Eight other microRNAs displayed significant expression changes over time in culture but did not clearly correlate with the LPS effect.
  • The researchers predicted certain immunity-associated target genes for miR-155, such as SOCS1, TAB2, and components of the PI3K signaling pathway. These predictions imply that miR-155 could influence the acute inflammatory response to LPS.
  • The researchers found substantial conservation of the miR-155 precursor gene and associated promoter regions between horses and humans.

Contribution to the Scientific Community

  • The research has charted both basal and LPS-induced microRNA expression patterns in horse leucocytes—a finding that aligns with patterns described in human cells.
  • This study can be pivotal for future research about the role of microRNAs in immune responses, particularly in horses. Additionally, it can also be an essential resource for interspecies comparative studies, mainly around how microRNAs contribute to the inflammatory response during sepsis and endotoxemia.

Cite This Article

APA
Parkinson NJ, Buechner-Maxwell VA, Witonsky SG, Pleasant RS, Werre SR, Ahmed SA. (2017). Characterization of basal and lipopolysaccharide-induced microRNA expression in equine peripheral blood mononuclear cells using Next-Generation Sequencing. PLoS One, 12(5), e0177664. https://doi.org/10.1371/journal.pone.0177664

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 12
Issue: 5
Pages: e0177664
PII: e0177664

Researcher Affiliations

Parkinson, Nicholas J
  • Department of Large Animal Clinical Sciences, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic and State University, Blacksburg, Virginia, United States of America.
Buechner-Maxwell, Virginia A
  • Department of Large Animal Clinical Sciences, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic and State University, Blacksburg, Virginia, United States of America.
Witonsky, Sharon G
  • Department of Large Animal Clinical Sciences, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic and State University, Blacksburg, Virginia, United States of America.
Pleasant, R Scott
  • Department of Large Animal Clinical Sciences, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic and State University, Blacksburg, Virginia, United States of America.
Werre, Stephen R
  • Laboratory for Study Design and Statistical Analysis, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic and State University, Blacksburg, Virginia, United States of America.
Ahmed, S Ansar
  • Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic and State University, Blacksburg, Virginia, United States of America.

MeSH Terms

  • Animals
  • Cytokines / biosynthesis
  • High-Throughput Nucleotide Sequencing
  • Horses
  • Leukocytes, Mononuclear / metabolism
  • Lipopolysaccharides / pharmacology
  • MicroRNAs / blood
  • MicroRNAs / genetics
  • Reverse Transcriptase Polymerase Chain Reaction

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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Citations

This article has been cited 4 times.
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