Gene expression changes in damaged osteoarthritic cartilage identify a signature of non-chondrogenic and mechanical responses S.L. Dunn, J. Soul, S.

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Gene expression changes in damaged osteoarthritic cartilage identify a signature of non-chondrogenic and mechanical responses S.L. Dunn, J. Soul, S. Anand, J.-M. Schwartz, R.P. Boot-Handford, T.E. Hardingham Osteoarthritis and Cartilage Volume 24, Issue 8, Pages 1431-1440 (August 2016) DOI: 10.1016/j.joca.2016.03.007 Copyright © 2016 The Authors Terms and Conditions

Fig. 1 Histological grading and GAG content of OA articular cartilage. Safranin O staining of the PLC (A) and DMC (B) of representative OA patient samples. Modified Mankin score of the articular cartilage (C) and GAG content of the cartilage tissue (D) obtained from the PLC and the DMC and used for RNA-seq analysis and an independent patient cohort for validation (n = 16). Images acquired using a [20×/0.80 Plan Apo] objective using the 3D Histech Pannoramic 250 Flash II slide scanner. Magnification× 5. ****P < 0.0001, **P < 0.01. Osteoarthritis and Cartilage 2016 24, 1431-1440DOI: (10.1016/j.joca.2016.03.007) Copyright © 2016 The Authors Terms and Conditions

Fig. 2 Comparison of intact vs damaged OA cartilage transcriptome studies. Overlap of DEGs identified by the RNA-seq data and the two existing microarray datasets Snelling et al. and Ramos et al. (A). The log2 fold change of the 22 DEGs in the three compared datasets (B). Osteoarthritis and Cartilage 2016 24, 1431-1440DOI: (10.1016/j.joca.2016.03.007) Copyright © 2016 The Authors Terms and Conditions

Fig. 3 PhenomeExpress analysis. Network analysis incorporating cross-species gene-phenotype associations, identified 23 differentially expressed networks based on direct protein–protein interactions in the damaged cartilage. Sub-networks linked to OA included; mitotic cell cycle (P = 0.0001) (A), regulation of transcription (P = 0.021) (B), Wnt signalling and calcium modulating pathway (P = 0.0102) (C), apoptotic processes (P = 0.0056) (D), negative regulation of blood coagulation (P = 0.0001) (E) and ECM organisation (P = 0.0001) (F). The fold change of the proteins is shown by the node colour and OA associated phenotype annotated proteins used to generate the sub-networks are shown with a black border. Osteoarthritis and Cartilage 2016 24, 1431-1440DOI: (10.1016/j.joca.2016.03.007) Copyright © 2016 The Authors Terms and Conditions

Fig. 4 Multi-layered architecture of damaged OA cartilage signalling. Network showing regulatory interactions between the differentially expressed, predicted upstream TFs and the PhenomeExpress identified pathways in damaged OA cartilage. Transcriptional links between the four upstream TFs are shown with grey arrows. Regulatory links between the TFs and significantly upregulated target genes present in PhenomeExpress pathways are shown red, green, pink and blue for FOSL1, AHR, E2F1 and FOXM1 respectively. The PhenomeExpress pathways are named by the top enriched GO biological process term. Only PhenomeExpress pathways with at least two target genes present are shown. Osteoarthritis and Cartilage 2016 24, 1431-1440DOI: (10.1016/j.joca.2016.03.007) Copyright © 2016 The Authors Terms and Conditions