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: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: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: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:This study investigated the systemic effects of 0.1 THz irradiation on the inflammatory status of rheumatoid arthritis (RA) in the collagen-induced arthritis (CIA) mouse model. It was found that THz irradiation apparently alleviated the symptoms of arthritis in CIA mice, reducing the joint swelling, mitigating the tissue damage and relieving the expression of pro-inflammatory factors (TNF-, IL-6, RANKL and MMPs) in the swollen joint of the CIA mice. Moreover, the reduction of serum inflammatory cytokines levels together with the reduced CD4+ T cell count and the recovered percentage of regulatory T cells (Treg) indicated that THz irradiation exerted a systemic immunoregulatory activity in CIA mice. The blood leukocyte mRNA-seq GO analysis also enriched several biological processes including the differentiation and adhesion of leukocytes, lymphocyte differentiation and T cell activation, providing additional supporting evidence for the immunoregulatory effects of THz.
Project description:The goal of this study is to define the pathophysiology of synovial fibroblasts in chroninc joint inflammation using NGS-derived synovial transcriptome profiling (RNA-seq) Methods: Synovial fibroblasts were isolated by FACs sorting [CD45-CD31-Popdoplanin+] from the synovium of healthy mice and mice undergoing experimental Collagen-Induced Arthritis (12 weeks old)(CIA). Methods: Whole Transcriptome Profiling of healthy and CIA synovial fibroblasts were generated by deep sequencing, in triplicate, using Illumina NextSeq™ 500 platform. Libraries were prepared using polyA selection (TruSeq stranded mRNA kit). Methods: The sequence reads that passed quality filters were aligned to mouse reference genome (GRCM38) using Hisat2 version 2.1.0. we mapped about 30 million sequence reads per sample (75 bp, paired-end) Methods: Featurecounts version 1.4.6 was used to quantify reads counts. Data quality control, non-expressed gene filtering, median ratio normalization (MRN) implemented in DESeq2 package and identification of differentially expressed (DE) genes were done using the R bioconductor project DEbrowser. Results: Results: We detected several differentially expressed (DE) genes in CIA synovial fibroblasts compared to naïve cells. These included 298 up-regulated genes and 88 down-regulated (>2 fold, adjp <0.05). DE genes reflected the reflecting the hyperplasia and activation observed during chronic joint disease. Pathways associated to DE genes were cell cycle and cell division and inflammatory response. Conclusions: our study shows pathophysiological changes associated to inflammatory synovial fibroblasts in a model of rheumatoid arthritis. Cells were directly isolated from fresh tissue, providing a valuable tool to study stromal inflammation.
Project description:The destruction of bone and cartilage results in a loss of joint functionality, critically impairing the quality of life in arthritis patients. Synovial fibroblasts (SFs) critically contribute to the pathogenesis of rheumatoid arthritis (RA) by acquiring either a pro-inflammatory or tissue-destructive phenotype. To explore the molecular mechanisms underlying the pathogenic fibroblast phenotype in arthritis, we performed single-cell RNA sequencing (scRNA-seq) on the synovial cells which were isolated from collagen-induced arthritis (CIA) mice.
Project description:Intermittent fasting (IF) reduces the inflammatory symptoms of Rheumatoid Arthritis (RA); however, its anti-inflammatory underlying molecular mechanisms are unknown. Aim: To investigate the effect of IF on joint inflammatory molecular mechanisms in collagen-induced arthritis (CIA) through DNA microarray and clinical and histopathological evaluations. Methods: CIA was induced in sixteen male DBA/1 mice divided into two groups, one received every other day IF for four weeks. The clinical, histological, and transcriptomic analyses, through DNA microarray, were done to evaluate the effect of IF on joint inflammation and remodeling. Results: CIA mice with IF show lower arthritis incidence and severity. IF also decreased the histological inflammation and joint damage. DNA microarrays dataset revealed 364 overexpressed and 543 subexpressed genes. The highest over- and subexpressed genes were the Ereg and MUPs, respectively. Retinol biosynthesis and the macrophage alternative activation pathways were associated with a positive z-score. Pathways with negative z-scores were related to the inflammatory process, including systemic lupus erythematosus in B cells and osteoarthritis, in which the RA-related genes Cd72, Cd79a, Ifna, Il33, and Bglap2 were found to be decreased. The bioinformatic algorithm identified four central possible regulators: CEBPA, FOXO1, HIF1A, PPARG, and PPARA. In the analysis of diseases and biofunctions, molecular transport, small molecule biochemistry, and lipid and carbohydrate metabolism tended to increase; conversely, organismal injury and abnormalities, inflammatory response, metabolic diseases, cancer, and endocrine, reproductive, gastrointestinal, skeletal, and muscular system disorders, were decreased by IF