Transcriptional Interactomic Inhibition of ROR[alpha] Suppresses Th17-Related Inflammation

Purpose: Th17 cells and their cytokines are implicated in the pathogenesis of various autoimmune diseases. Retinoic acid-related orphan receptor alpha (ROR[alpha]) is a transcription factor for the differentiation and the inflammatory functions of Th17 cells. In this study, we generated the nucleus-transducible form of transcription modulation domain of ROR[alpha] (nt-ROR[alpha]-TMD) to investigate the functional roles of ROR[alpha] in vitro and in vivo under normal physiological condition without genetic alteration. Methods: The functions of nt-ROR[alpha]-TMD were analyzed in vitro through flow cytometry, luciferase assay, ELISA, and transcriptome sequencing. Finally, the in vivo therapeutic effects of nt-ROR[alpha]-TMD were verified in dextran sulfate sodium (DSS)-induced colitis mice. Results: nt-ROR[alpha]-TMD was effectively delivered into the cell nucleus in a dose- and time-dependent manner without any cellular toxicity. nt-ROR[alpha]-TMD competitively inhibited the ROR[alpha]-mediated transcription but not ROR[gamma]t-mediated transcription. Secretion of IL-17A from the splenocytes was suppressed by nt-ROR[alpha]-TMD without affecting the secretion of Th1- or Th2-type cytokine and T cell activation events such as induction of CD69 and CD25. The differentiation potential of naive T cells into Th17 cells, not into Th1, Th2, or Treg cells, was significantly blocked by nt-ROR[alpha]-TMD. Consistently, mRNA sequencing analysis showed that nt-ROR[alpha]-TMD treatment down-regulated the expression of the genes related to the differentiation and functions of Th17 cells. Treatment of DSS-induced colitis mice with nt-ROR[alpha]-TMD improved the overall symptoms of colitis, such as body weight change, colon length, infiltration of inflammatory cells, and the level of inflammatory cytokines in the serum. In the mesenteric lymph node (MLN) of the nt-ROR[alpha]-TMD-treated mice, the population of CD4+IL-17A+ Th17 cells was reduced, and the population of CD4+Foxp3+ Treg cells increased. Conclusion: nt-ROR[alpha]-TMD has a potential to be developed as a novel therapeutic reagent for treating various inflammatory diseases in which Th17 cells are the leading pathological player. Keywords: retinoic acid-related orphan receptor, T helper 17, transcriptional modulation, DSS-induced colitis, inflammatory disease