TYK2 inhibition reduces type 3 immunity and modifies disease progression in murine spondyloarthritis
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
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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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 6

TYK2 inhibition by small molecule and genetic mutation suppresses IL-23–induced activation of murine T cells in vitro.

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TYK2 inhibition by small molecule and genetic mutation suppresses IL-23–...
Lymph node cells were stimulated in vitro, and γδ T cell activation was assessed by flow cytometry. (A and B) Lymphocytes were preincubated with NDI-031407 for 30 minutes before stimulation for 15 minutes with 400 ng/mL IL-23. (A) Representative plots showing p-STAT3 staining under IL-23 or pervanadate stimulation in αβ and γδ T cells. (B) Pooled data of NDI-031407–treated, IL-23–stimulated γδ T cells. (C and D) Representative plots and pooled data of p-STAT3 in γδ T cells under IL-23 stimulation in TYK2 kinase-dead mice (TYK2K923E). (E–H) Lymphocytes were stimulated with 10 ng/mL IL-1β and/or 20 ng/mL IL-23 with brefeldin A for 4.5 hours before detection of IL-17A/IL-22 in γδ T cells by flow cytometry. Where applicable, lymphocytes were treated with NDI-031407 for 30 minutes before stimulation. (E) Representative plots showing cytokine staining in γδ T cells. (F) Pooled data of IL-17A+ and IL-22+ γδ T cells treated with NDI-031407. (G and H) IL-23/IL-1β stimulation of TYK2K923E (G) and TYK2–/– (H) lymphocytes. (I) IL-23/IL-1β stimulation of TYK2K923E lymphocytes with pan-JAK inhibitor ruxolitinib. Data are mean ± SEM whereby each data point is a separate well. All data are from a single experiment representative of 2–3 independent experiments. D and G, t test with Welch’s correction; B and F, 1-way ANOVA with Dunnett’s post hoc test compared with cytokine-stimulated/vehicle control; H, 1-way ANOVA with Dunnett’s post hoc test. In I, IL-1β only was compared with IL-1β/IL-23/1000 nM ruxolitinib for each genotype by t test with Welch’s correction, and 2-way ANOVA was used to compare all IL-1β/IL-23–stimulated cells and Dunnett’s multiple-comparisons test to compare vehicle- vs. ruxolitinib-treated samples within each genotype. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.