Ruxolitinib inhibits IFNγ-stimulated Sjögren’s salivary gland MSC HLA-DR expression and chemokine-dependent T cell migration

Abstract

Objectives Sjӧgren’s disease (SjD) is a systemic autoimmune disease characterized by focal lymphocytic infiltrate of salivary glands (SGs) and high SG IFNγ, both of which are associated with elevated lymphoma risk. IFNγ is also biologically relevant to mesenchymal stromal cells (MSCs), a SG resident cell with unique niche regenerative and immunoregulatory capacities. In contrast to the role of IFNγ in SjD, IFNγ promotes an anti-inflammatory MSC phenotype in other diseases. The objective of this study was to define the immunobiology of IFNγ-exposed SG-MSCs with and without the JAK1 & 2 inhibitor, ruxolitinib. Methods SG-MSCs were isolated from SjD and controls human subjects. SG-MSCs were treated with 10 ng/ml IFNγ +/- 1000 nM ruxolitinib. Experimental methods included flow cytometry, RNA-sequencing, chemokine array, ELISA, and transwell chemotaxis experiments. Results We found that IFNγ promoted expression of SG-MSC immunomodulatory markers, including HLA-DR, and this expression was inhibited by ruxolitinib. We confirmed the differential expression of CXCL9, CXCL10, CXCL11, CCL2, and CCL7, initially identified with RNA-sequencing. SG-MSCs promoted CD4+ T cell chemotaxis when pre-stimulated with IFNγ. Ruxolitinib blocks chemotaxis through inhibition of SG-MSC production of CXCL9, CXCL10, and CXCL11. Conclusions These findings establish that ruxolitinib inhibits IFNγ-induced expression of SG-MSC immunomodulatory markers and chemokines. Ruxolitinib also reverses IFNγ-induced CD4+ T cell chemotaxis, through inhibition of CXCL9, -10, and -11. Because IFNγ is higher in SjD than control SGs, we have identified SG-MSCs as a plausible pathogenic cell type in SjD. We provide proof of concept supporting further study of ruxolitinib to treat SjD.

CD169+ classical monocyte as an important participant in Graves’ ophthalmopathy through CXCL12-CXCR4 axis

“…This drug has recently shown promising results in clinical studies for treating autoimmune diseases and fibrotic lesions. 55 , 56 , 57 Our results showed that ruxolitinib treatment significantly decreased IFN-γ-stimulated CD169 + clas_mono, both in terms of transcriptional and protein levels ( Figure 5 G). Moreover, we observed a significant improvement in survival analysis among patients with a low proportion of CD169 + classical monocytes ( Figure 5 F).…”

“…Of significance, CD169 + clas_mono responded positively to ruxolitinib, a highly selective JAK inhibitor that is effective and safe for treating autoimmune diseases and fibrotic lesions. 55 , 56 , 57 …”

Emerging biologic frontiers for Sjogren’s syndrome: Unveiling novel approaches with emphasis on extra glandular pathology

“…Notable, promising research by Renaudineau and colleagues suggests that JAK1/2 inhibitors, AG490 and ruxolitinib, may reverse Frontiers in Pharmacology frontiersin.org 13 specific pathways implicated in pSS pathogenesis (McCoy et al, 2022). Tofacitinib, a JAK1 and JAK3 inhibitor, has the potential to treat pSS by restoring autophagy and mitigating inflammation, particularly by targeting IL-6 expression (Barrera et al, 2021).…”

JAK/STAT pathway targeting in primary Sjögren syndrome

“…The biological effects of ruxolitinib, a JAK1 and JAK2 inhibitor, on mesenchymal stromal cells (MSCs), isolated from salivary glands of both pSS patients and controls, have been recently evaluated in vitro . [ 76 ] A ruxolitinib-mediated inhibition of IFN-γ-induced expression of MSC immunomodulatory markers, such as HLA-DR (Major histocompatibility complex (MHC) II cell surface receptor) expression, has emerged by these experimental studies, together with the block of CD4 + T cell chemotaxis through the inhibition of MSC production of CXCL9, CXCL10, and CXCL11, suggesting potential implications for ruxolitinib in pSS therapy. [ 76 ]…”