miR-146a in Myasthenia Gravis Thymus Bridges Innate Immunity With Autoimmunity and Is Linked to Therapeutic Effects of Corticosteroids

• Published in: Frontiers in immunology 2020-03, Vol.11, p.142
• Main Authors: Bortone, Federica, Scandiffio, Letizia, Marcuzzo, Stefania, Bonanno, Silvia, Frangiamore, Rita, Motta, Teresio, Antozzi, Carlo, Mantegazza, Renato, Cavalcante, Paola, Bernasconi, Pia
• Format: Article
• Language: English
• Subjects: Adolescent; Adrenal Cortex Hormones - pharmacology; Adrenal Cortex Hormones - therapeutic use; Adult; Autoimmunity; B-Lymphocytes - immunology; Cells, Cultured; Child; Female; Gene Expression Regulation, Neoplastic - drug effects; Germinal Center - drug effects; Germinal Center - immunology; Humans; Immunity, Innate; Immunology; innate immunity; Male; MicroRNAs - blood; MicroRNAs - genetics; Middle Aged; miR-146a; myasthenia gravis; Myasthenia Gravis - blood; Myasthenia Gravis - drug therapy; Myasthenia Gravis - immunology; Prednisone - pharmacology; Prednisone - therapeutic use; RNA, Messenger - genetics; Signal Transduction - drug effects; thymus; Thymus Gland - immunology; Toll-Like Receptors - metabolism; Treatment Outcome; Young Adult
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2020.00142

Toll-like receptor (TLR)-mediated innate immune responses are critically involved in the pathogenesis of myasthenia gravis (MG), an autoimmune disorder affecting neuromuscular junction mainly mediated by antiacetylcholine receptor antibodies. Considerable evidence indicate that uncontrolled TLR activation and chronic inflammation significantly contribute to hyperplastic changes and germinal center (GC) formation in the MG thymus, ultimately leading to autoantibody production and autoimmunity. miR-146a is a key modulator of innate immunity, whose dysregulation has been associated with autoimmune diseases. It acts as inhibitor of TLR pathways, mainly by targeting the nuclear factor kappa B (NF-κB) signaling transducers, interleukin 1 receptor associated kinase 1 (IRAK1) and tumor necrosis factor (TNF) receptor associated factor 6 (TRAF6); miR-146a is also able to target c-REL, inducible T-cell costimulator (ICOS), and Fas cell surface death receptor (FAS), known to regulate B-cell function and GC response. Herein, we investigated the miR-146a contribution to the intrathymic MG pathogenesis. By real-time PCR, we found that miR-146a expression was significantly downregulated in hyperplastic MG compared to control thymuses; contrariwise, IRAK1, TRAF6, c-REL, and ICOS messenger RNA (mRNA) levels were upregulated and negatively correlated with miR-146a levels. Microdissection experiments revealed that miR-146a deficiency in hyperplastic MG thymuses was not due to GCs, but restricted to the GC-surrounding medulla, characterized by IRAK1 overexpression. We also showed higher c-REL and ICOS mRNA levels, and lower FAS mRNA levels, in GCs than in the remaining medulla, according to the contribution of these molecules in GC formation. By double immunofluorescence, an increased proportion of IRAK1-expressing dendritic cells and macrophages was found in hyperplastic MG compared to control thymuses, along with GC immunoreactivity for c-REL. Interestingly, in corticosteroid-treated MG patients intrathymic miR-146a and mRNA target levels were comparable to those of controls, suggesting that immunosuppressive therapy may restore the microRNA (miRNA) levels. Indeed, an effect of prednisone on miR-146a expression was demonstrated on peripheral blood cells. Serum miR-146a levels were lower in MG patients compared to controls, indicating dysregulation of the circulating miRNA. Our overall findings strongly suggest that defective miR-146a expression could contribute to persistent