MiR429 regulates IL‐17A expression in inflammatory monocytes in Rheumatoid Arthritis

Abstract

Rheumatoid Arthritis (RA) is a chronic, autoimmune disorder that leads to inflammation and degradation of joints. IL17A, a proinflammatory cytokine, has been previously shown to be involved in migratory inflammation pathways in autoimmune disorders like RA. IL17A initiates RA pathogenesis, suggesting that treatments targeted to neutralize IL17A would be effective therapies. Currently, anti‐IL‐17 therapies are among the most efficacious for treating RA. Additionally, studies indicate that monocytes are also causative agents of inflammation in RA. microRNAs (miRNA) have been identified as translational inhibitors in mRNA of eukaryotic cells. The relationship among monocytes, miRNAs and mRNA must be better analyzed in order to depict inflammatory pathways involved in RA. miR429 was chosen in this experiment because previous research in our lab has suggested that it is upregulated in RA monocytes. We hypothesized that the binding of miR429 to the 3′ UTR region of IL17A would down regulate the expression of IL17A and thus inflammation.Methods and ResultsGene targets involved in monocytes and inflammation were identified for miR429 in the miRNA databases miRanda and mirSVR. miR429 was found to have a relatively high binding score with IL17A. Isolating the 3′ UTR of the IL17A target gene was the first step in our series of experiments. U937 monocytes were grown and cDNA was created from isolated RNA. Primers were then designed using Primer‐BLAST program, and were created based on the hypothetical binding site on the IL17A 3′ UTR. PCR was used to amplify the 3′ UTR and a restriction enzyme digest was performed on the insert DNA and the pMirGlo vector.ConclusionOur results suggest that IL17A and miR429 are expressed by monocytes and may interact to modulate inflammation in RA.Support or Funding InformationAlbion College Biology Department and the Foundation for Undergraduate Research, Scholarly, and Creative ActivityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.