한빛사논문
Saseong Lee,1 Eunbyeol Choi,1,2 Sehyun Chae,3 Jung Hee Koh,1,4 Yoolim Choi,5 Jung Gon Kim ,1,6 Seung-Ah Yoo,1,7 Daehee Hwang,5 Wan-Uk Kim 1,4
1Center for Integrative Rheumatoid Transcriptomics and Dynamics, The Catholic University of Korea, Seoul, The Republic of Korea
2Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul, The Republic of Korea
3Neurovascular Unit Research Group, Korea Brain Research Institute, Daegu, The Republic of Korea
4Department of Internal Medicine, The Catholic University of Korea, School of Medicine, Seoul, The Republic of Korea
5Department of Biological Sciences, Seoul National University, Seoul, The Republic of Korea
6Department of Internal Medicine, Inje University Ilsan Paik Hospital, Goyang, The Republic of Korea
7Department of Medical Life Sciences, The Catholic University of Korea, Seoul, The Republic of Korea
SL, EC and SC are joint first authors.
Correspondence to Professor Wan-Uk Kim
Abstract
Objectives: 'Invasive pannus' is a pathological hallmark of rheumatoid arthritis (RA). This study aimed to investigate secretome profile of synovial fibroblasts of patients with RA (RA-FLSs), a major cell type comprising the invasive pannus.
Methods: Secreted proteins from RA-FLSs were first identified using liquid chromatography-tandem mass spectrometry analysis. Ultrasonography was performed for affected joints to define synovitis severity at the time of arthrocentesis. Expression levels of myosin heavy chain 9 (MYH9) in RA-FLSs and synovial tissues were determined by ELISA, western blot analysis and immunostaining. A humanised synovitis model was induced in immuno-deficient mice.
Results: We first identified 843 proteins secreted from RA-FLSs; 48.5% of the secretome was associated with pannus-driven pathologies. Parallel reaction monitoring analysis of the secretome facilitated discovery of 16 key proteins related to 'invasive pannus', including MYH9, in the synovial fluids, which represented synovial pathology based on ultrasonography and inflammatory activity in the joints. Particularly, MYH9, a key protein in actin-based cell motility, showed a strong correlation with fibroblastic activity in the transcriptome profile of RA synovia. Moreover, MYH9 expression was elevated in cultured RA-FLSs and RA synovium, and its secretion was induced by interleukin-1β, tumour necrosis factor α, toll-like receptor ligation and endoplasmic reticulum stimuli. Functional experiments demonstrated that MYH9 promoted migration and invasion of RA-FLSs in vitro and in a humanised synovitis model, which was substantially inhibited by blebbistatin, a specific MYH9 inhibitor.
Conclusions: This study provides a comprehensive resource of the RA-FLS-derived secretome and suggests that MYH9 represents a promising target for retarding abnormal migration and invasion of RA-FLSs.
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