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BS25 Investigating the MIR-101-3P/TRIB1 axis in macrophage immunometabolism
  1. Chiara Niespolo1,
  2. Juan Salamanca Viloria2,3,
  3. Sumeet Deshmukh4,
  4. Oscar Villacanas Perez2,
  5. Ian Sudbery4,
  6. Heather Wilson1,
  7. Endre Kiss-Toth1
  1. 1Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield
  2. 2Mind the Byte (formerly Intelligent Pharma), Barcelona
  3. 3University of Barcelona
  4. 4Department of Molecular Biology and Biotechnology, University of Sheffield


Introduction MiR-101-3p has been implicated in the regulation of macrophage cholesterol efflux and pro-inflammatory genes by directly targeting ABCA1 and MKP-1, which are both downregulated in atherosclerosis. By using bioinformatic analysis we identified TRIB1 as a direct target of miR-101. The TRIB1 gene plays major roles in myeloid cells, regulating inflammation and lipid metabolism. Its deficiency is associated with a severe reduction of anti-inflammatory tissue macrophages (M2-like cells) and an increase in plasma triglycerides.

Here we aimed to uncover the role of miR-101-3p/TRIB1 interaction and its consequences on human macrophage immunometabolism.

Methods MiRanda target prediction algorithm was used to identify macrophage-specific miRNAs targeting TRIB1, along with additional web-based tools. MiR-101-3p was selected based on its conservation among species as well as its high predictive score and free energy. A luciferase reporter assay was used to validate the miR-101-3p/TRIB1 interaction, employing a miR-101 mimic and inhibitor. Gene expression (RT-qPCR) and protein analysis (Western Blot) was carried out in human monocyte-derived macrophages (hMDMs) following transient manipulation of TRIB1 and miR-101 levels.

Results miR-101-3p has a functional binding site for the 3’UTR of TRIB1 and when overexpressed in hMDMs is able to reduce TRIB1 expression at both mRNA and protein levels. Over-expression of TRIB1 in hMDMs alters the levels of pro-inflammatory genes and scavenger receptors, inducing an M2-like phenotype. Conversely, overexpression of miR-101 is associated with a pro-inflammatory phenotype (M1-like).

Conclusion miR-101-3p offers a potential target to simultaneously enhance the expression of ABCA1, MKP-1 and TRIB1, thus improving macrophage lipid metabolism and opposing inflammation, thereby attenuating atherosclerosis development.

Conflict of interest No

  • macrophages
  • microRNAs
  • Tribbles-1

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