Differential modulation of Ahr and Arid5a: A promising therapeutic strategy for autoimmune encephalomyelitis

Multiple sclerosis (MS) is an autoimmune disease that involves demyelination of axons in the central nervous system (CNS) and affects patients worldwide. It has been demonstrated that ligand-activated Aryl Hydrocarbon Receptor (Ahr) ameliorates experimental autoimmune encephalomyelitis (EAE), a murine model of MS, by increasing CD4⁺FoxP3⁺ T cells. Recent evidence indicates that AT-rich interactive domain-containing protein 5a (Arid5a) is required for EAE pathogenesis by stabilizing Il6 and OX40 mRNAs. However, the differential modulation of Ahr and Arid5a in autoimmunity as a therapeutic strategy is unexplored. Herein, an in silico, in vitro and in vivo approach identified Flavipin (3,4,5-trihydroxy-6-methylphthalaldehyde) as an Ahr agonist that induces the expression of Ahr downstream genes in mouse CD4⁺ T cells and CD11b⁺ macrophages. Interestingly, Flavipin inhibited the stabilizing function of Arid5a and its counteracting effects on Regnase-1 on the 3' untranslated region (3'UTR) of target mRNAs. Furthermore, it inhibited the stabilizing function of Arid5a on Il23a 3'UTR, a newly identified target mRNA. In EAE, Flavipin ameliorated disease severity, with reduced CD4⁺IL-17⁺ T cells, IL-6 and TNF-α and increased CD4⁺FoxP3⁺ T cells. Moreover, EAE amelioration was concomitant with reduced CD4⁺OX40⁺ and CD4⁺CD45⁺ T cells in the CNS. RNA interference showed that the modulatory effects of Flavipin on pro- and anti-inflammatory mediators in CD4⁺ T cells and macrophages were Ahr- and/or Arid5a-dependent. In conclusion, our findings reveal differential modulation of Ahr and Arid5a as a new therapeutic strategy for MS.

3′UTR, 3′ untranslated region; ActinD, actinomycin D; Ahr; Ahr, aryl hydrocarbon receptor; Arid5a; Arid5a, AT-rich interactive domain-containing protein 5a; Arnt, Ahr nuclear translocator; Autoimmunity; CFA, complete Freund's adjuvant; CNS, central nervous system; EAE, experimental autoimmune encephalomyelitis; Inflammation; LPS, lipopolysaccharide; MOG35-55, myelin oligodendrocyte glycoprotein; MS, multiple sclerosis; Multiple sclerosis; PAS-A and PAS-B, Per-Arnt-Sim domain; RBP, RNA-binding protein; RIP, RNA immunoprecipitation; SPF, specific pathogen-free; Therapeutic; miR, microRNA.