Project description:Knee joint synovium was used for gene expression analysis of mouse collagen induced arthritis (CIA). Synovium was prepared at day 30 after initial sensitization from: healthy controls, CIA animals with no, with mild, with moderate, or with severe joint inflammation. Each sample group is represented by three replicates, each consisting of tissue collected from three to four animals. Experiment Overall Design: The data set consists of 15 samples: five groups with three replicates each. One sample group is from healthy controls, the other groups are from CIA animals with different degress of joint inflammation.
Project description:Knee joint synovium was used for gene expression analysis of mouse collagen induced arthritis (CIA). Synovium was prepared at day 30 after initial sensitization from: healthy controls, CIA animals with no, with mild, with moderate, or with severe joint inflammation. Each sample group is represented by three replicates, each consisting of tissue collected from three to four animals. Keywords: disease severity analysis
Project description:Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation, immune cell infiltration, and progressive joint destruction. Commonly used mouse models each carry practical constraints: collagen-induced arthritis (CIA) is largely restricted to the DBA/1 strain, whereas collagen antibody-induced arthritis (CAIA) is limited by antibody cost. To combine strain flexibility with a robust and reproducible effector phase, we established collagen- and antibody-induced arthritis (C&AIA) on the C57BL/6 background. To characterize the synovial cellular landscape of this model, we performed droplet-based single-cell RNA sequencing on synovial tissue dissected from inflamed knee joints of arthritic C&AIA mice. This dataset resolves the immune and stromal compartments of the C&AIA synovium and enables cross-species comparison with human RA synovial single-cell data.
Project description:Collagen-induced arthritis (CIA) was established in DBA/1 male mice using bovine type II collagen emulsified in Freund’s adjuvant. Arthritis severity was scored by an arthritis index, and CIA-positive animals were randomized to a CIA group or a CIA+F20 group (10 mg/kg,p.o. once daily for 21 days); age-matched non-immunized mice served as healthy controls. At endpoint, both forelimbs were collected by cutting at the elbow level to retain the forearm, wrist, and paw. Skin and excess muscle were removed while preserving joint capsules, synovium, and periarticular tissues for RNA-seq. Total RNA was extracted, mRNA libraries were prepared and sequenced (paired-end) on an MGI platform. Clean reads were obtained with fastp, aligned to Mus_musculus.GRCm39.109 (Ensembl) using HISAT2, and quantified with featureCounts.
Project description:In rheumatoid arthritis (RA), inflammation and joint destruction are exacerbated by a complicated interaction among immune cells, synovial fibroblasts and bone cells. It remains to be elucidated which cell-cell interaction critically drives the pathogenesis of RA and serves as a therapeutic target for synthetic disease modifying antirheumatic drugs (DMARDs) such as janus kinase (JAK) inhibitors. we performed a scRNA-seq analysis of the synovium of collagen-induced arthritis (CIA) mice treated with JAK inhibitor, followed by a computational analysis to identify the drug target cells and signaling pathways in vivo
Project description:High expression alleles of the innate cytokine, macrophage migration inhibitory factor (MIF), are associated with the development or the severity of autoimmune inflammatory diseases, including rheumatoid arthritis. Numerous studies support MIF’s role in activating inflammatory pathways and MIF inhibition reduces joint pathology in different experimental models of arthritis. We examined the impact of gene deletion of MIF or its cognate receptor CD74 in the T cell-dependent model of collagen-induced arthritis (CIA) and observed the complete absence of arthritis development, suggesting an unforeseen role for MIF/CD74 signaling in the development of arthritogenic T cells. While MIF has been shown in model systems to contribute to T cell activation by augmenting innate responses, fewer than 1% of T lineage cells express CD74 in naïve spleens and lymph nodes, and its functional consequences in pathogenic T cell subpopulations have not been studied. We found CD74+ T cells to expand during CIA and to increase in number within joint synovium, where they express an effector memory phenotype and recapitulate CIA development upon transfer into naïve mice. We further found evidence for the presence of CD74+ T cells in the circulation and joint synovium of patients with rheumatoid arthritis. MIF-dependent, CD74+ T cells may contribute to the chronicity of rheumatoid synovitis and to disease relapse in previously inflamed joints.
Project description:Fibroblast-like synoviocytes (FLS) were isolated from the inflamed joints of mice with collagen-induced arthritis (CIA) or the joints of naive littermates to characterise gene expression changes in response to chronic inflammation. Cells were isolated at two times of day (zeitgeber time, ZT4 and ZT16) to characterise diurnal variation in inflammatory symptoms and responses. FLS cells were isolated from joint digests based on Podoplanin expression prior to RNA extraction and library preparation for sequencing.
Project description:Oxylipins play important roles in various biological processes and are considered as mediators of inflammation for a wide range of diseases such as rheumatoid arthritis (RA). The purpose of this research was to study differences in oxylipin levels between a widely used collagen-induced arthritis (CIA) mice model and healthy control (Ctrl) mice. DBA/1J male mice (age: 6-7 weeks) were selected and randomly divided into two groups, viz. a CIA- and a Ctrl group. The CIA mice were injected intraperitoneal (i.p.) with the joint cartilage component collagen type II (CII) and an adjuvant injection of lipopolysaccharide (LPS). Oxylipin metabolites were extracted from plasma for each individual sample using solid phase extraction (SPE) and were detected with high performance liquid chromatography/tandem mass spectrometry (HPLC-ESI-MS/MS), using dynamic multiple reaction monitoring (dMRM). Both univariate and multivariate statistical analysis was applied. The results in univariate student's t-test revealed 10 significantly up- or down-regulated oxylipins in CIA mice, which were supplemented by another 6 additional oxylipins, contributing to group clustering upon multivariate analysis. The dysregulation of these oxylipins revealed the presence of ROS-generated oxylipins and an increase of inflammation in CIA mice. The results also suggested that the Collagen-induced arthritis might associate with dysregulation of apoptosis, possibly inhibited by activated NF- κ B because of insufficient PPAR-γ ligands.
Project description:Rheumatoid arthritis (RA) induced destruction of knee joints is a common cause of total knee arthroplasty (TKA). Under single cell RNA sequencing generated by 10x Genomics, we identified CD142+ synovial fibroblasts as a novel cluster, located at the sublining layer in normal and osteoarthritis knee synovium, but elevated and distributed at the lining layer in RA knee synovium, and infiltrated in multiple RA joint synovia. Intra-articular injection of CD142+ synovial fibroblasts can quickly and drastically damage the meniscus but has a slight effect on cartilage.Long-term follow-up of the RA cohort indicated that enriched CD142+ synovial fibroblasts in the lining layer was a risk factor for severe joint destruction and eventually underwent TKA. Our results demonstrate that CD142+ synovial fibroblasts can be used as an indicator to assess prognosis and a therapeutic target to inhibit meniscal damage, thereby alleviating RA knee joint destruction.