Dynamic transcriptome analysis unveils key proresolving factors of chronic inflammatory arthritis
Jin-Sun Kong, … , Daehee Hwang, Wan-Uk Kim
Jin-Sun Kong, … , Daehee Hwang, Wan-Uk Kim
Published May 14, 2020
Citation Information: J Clin Invest. 2020;130(8):3974-3986. https://doi.org/10.1172/JCI126866.
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Research Article Autoimmunity Inflammation

Dynamic transcriptome analysis unveils key proresolving factors of chronic inflammatory arthritis

Abstract

Despite recent advances in understanding chronic inflammation remission, global analyses have not been explored to systematically discover genes or pathways underlying the resolution dynamics of chronic inflammatory diseases. Here, we performed time-course gene expression profiling of mouse synovial tissues along progression and resolution of collagen-induced arthritis (CIA) and identified genes associated with inflammation resolution. Through network analysis of these genes, we predicted 3 key secretory factors responsible for the resolution of CIA: Itgb1, Rps3, and Ywhaz. These factors were predominantly expressed by Tregs and antiinflammatory M2 macrophages, suppressing production of proinflammatory cytokines. In particular, Ywhaz was elevated in the sera of mice with arthritis resolution and in the urine of rheumatoid arthritis (RA) patients with good therapeutic responses. Moreover, adenovirus-mediated transfer of the Ywhaz gene to the affected joints substantially inhibited arthritis progression in mice with CIA and suppressed expression of proinflammatory cytokines in joint tissues, lymph nodes, and spleens, suggesting Ywhaz is an excellent target for RA therapy. Therefore, our comprehensive analysis of dynamic synovial transcriptomes provides previously unidentified antiarthritic genes, Itgb1, Rps3, and Ywhaz, which can serve as molecular markers to predict disease remission, as well as therapeutic targets for chronic inflammatory arthritis.

Authors

Jin-Sun Kong, Ji-Hwan Park, Seung-Ah Yoo, Ki-Myo Kim, Yeung-Jin Bae, Yune-Jung Park, Chul-Soo Cho, Daehee Hwang, Wan-Uk Kim

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Figure 7

Expression of Ywhaz and GFP after intra-articular injection of Ad-Ywhaz tagged with Gfp in mice with CIA.

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Expression of Ywhaz and GFP after intra-articular injection of Ad-Ywhaz ...
(A) A schematic diagram illustrating the establishment of CIA and treatment of mice with adenoviral vectors. Following primary and secondary immunization on days 0 and 14, respectively, 1 × 108 plaque-forming units (PFU) of the adenoviral vector containing Ywhaz (Ad-Ywhaz) or the control adenoviral vector (Ad-Con) in 10 μL HEPES were injected into the ankle joints of the mice at 30 and 37 days. Mice were then sacrificed at 33 or 52 days for further immunopathologic analyses. (B and C) mRNA expression levels of Ywhaz, Il6, and Tnf (B) and protein expression levels of Ywhaz (C) in the synovial tissues of CIA mice 33 days after primary immunization, which were determined by qRT-PCR and Western blot analysis, respectively. mRNA and protein levels of the target genes were normalized to those of Gapdh and β-actin, respectively. (D and E) GFP imaging (D) and qRT-PCR assays for Gfp (E) performed in the lung, liver, and ankle joint of CIA mice at 24, 48, and 72 hours after intra-articular injection of Ad-Ywhaz tagged with Gfp or those of mice without the injection. The color bar in D represents the gradient of radiant efficiency. Data in D are representative of 2 independent experiments with similar results, and data in B, C, and E are the mean ± SEM of more than 3 mice. For qRT-PCR assays, Gfp levels at each time point were first normalized to Gapdh levels, and then further normalized to the mean mRNA expression levels measured in the lung with intra-articular injection of Ad-Ywhaz tagged with Gfp. *P < 0.05, **P < 0.01, ***P < 0.001 as determined by Student’s t test (B and C) or by Kruskal-Wallis test with a post hoc test (Dunn’s correction; E).