TYK2 inhibition reduces type 3 immunity and modifies disease progression in murine spondyloarthritis
Eric Gracey, … , Wenyan Miao, Robert D. Inman
Eric Gracey, … , Wenyan Miao, Robert D. Inman
Published March 9, 2020
Citation Information: J Clin Invest. 2020;130(4):1863-1878. https://doi.org/10.1172/JCI126567.
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Research Article Autoimmunity

TYK2 inhibition reduces type 3 immunity and modifies disease progression in murine spondyloarthritis

Abstract

Spondyloarthritis (SpA) represents a family of inflammatory diseases of the spine and peripheral joints. Ankylosing spondylitis (AS) is the prototypic form of SpA in which progressive disease can lead to fusion of the spine. Therapeutically, knowledge of type 3 immunity has translated into the development of IL-23– and IL-17A–blocking antibodies for the treatment of SpA. Despite being able to provide symptomatic control, the current biologics do not prevent the fusion of joints in AS patients. Thus, there is an unmet need for disease-modifying drugs. Genetic studies have linked the Janus kinase TYK2 to AS. TYK2 is a mediator of type 3 immunity through intracellular signaling of IL-23. Here, we describe and characterize a potentially novel small-molecule inhibitor of TYK2 that blocked IL-23 signaling in vitro and inhibited disease progression in animal models of SpA. The effect of the inhibitor appears to be TYK2 specific, using TYK2-inactive mice, which further revealed a duality in the induction of IL-17A and IL-22 by IL-23. Specifically, IL-22 production was TYK2/JAK2/STAT3 dependent, while IL-17A was mostly JAK2 dependent. Finally, we examined the effects of AS-associated TYK2 SNPs on TYK2 expression and function and correlated them with AS disease progression. This work provides evidence that TYK2 inhibitors have great potential as an orally delivered therapeutic for SpA.

Authors

Eric Gracey, Dominika Hromadová, Melissa Lim, Zoya Qaiyum, Michael Zeng, Yuchen Yao, Archita Srinath, Yuriy Baglaenko, Natalia Yeremenko, William Westlin, Craig Masse, Mathias Müller, Birgit Strobl, Wenyan Miao, Robert D. Inman

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Figure 1

TYK2 inhibition by a novel small molecule blocks IL-23–induced STAT3 phosphorylation and IL-17A production in human CD4+ T cells.

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TYK2 inhibition by a novel small molecule blocks IL-23–induced STAT3 pho...
(A and B) NDI-031407, a novel TYK2 inhibitor, was tested for: (A) Specificity for TYK2 against JAK1–3 kinases by radiometric assay with peptide substrates. Activity represents the ratio of activated substrate in DMSO versus inhibitor treatment. (B) Potency for IL-12–induced p-STAT4 and GM-CSF–induced p-STAT5 in PMBCs and IL-12–induced IFN-γ in NK92 cells. Activity represents the ratio of p-STAT to total STAT. Data in A and B are from a single experiment, representative of 3 independent experiments. The horizontal lines represent 50% inhibition. (C) Magnetically purified CD4+ T cells were cultured with anti-CD2/CD3/CD28 beads for 3 days with NDI-031407 in the presence of 20 ng/mL of cytokines. At endpoint, IL-17A was assessed in the culture supernatant by ELISA. (D–G) PBMCs were stimulated for 4 days with anti-CD2/CD3/CD28 beads. Cells were then serum-starved and pretreated with JAKinib for 30 minutes before 15-minute stimulation with pervanadate, 400 ng/mL IL-6, or 400 ng/mL IL-23. STAT phosphorylation was assessed by flow cytometry. (D) Representative dot plots showing p-STAT3 in relation to mature CD4+ T cells (left) and representative gating for p-STAT3+ cells in mature CD4+ T cells with the indicated treatments (right). (E) Pooled data showing p-STAT3 in mature CD4+ T cells. (F and G) Comparison of NDI-031407, tofacitinib, and ruxolitinib inhibition of IL-23R and IL-6R. Representative histograms show p-STAT3 in mature CD4+ T cells: unstimulated (black dashed line), cytokine-stimulated (gray shading), or 50 nM (thin lines) and 500 nM (thick lines) of the respective JAKinib. Threshold used to gate p-STAT3+ (blue dashed line) and percentage positive are indicated in parentheses. Graph title indicates the cytokine-associated JAKs. (C and E) IL-6/vehicle vs. IL-6/500 nM NDI-031407 by Wilcoxon matched-pairs signed-rank test and stimulated/vehicle-treated wells vs. stimulated/NDI-031407–treated wells by paired 1-way ANOVA with Dunnett’s post hoc test comparing treatments with vehicle control. For all scatter plots, each point represents an independent donor. **P < 0.01, ***P < 0.001, ****P < 0.0001.