Dataset Information

A COMPREHENSIVE AND QUANTIFIABLE INSIGHT INTO THE PROTEOMES OF ARTICULAR CARTILAGE AND SUBCHONDRAL BONE

ABSTRACT: In this study, we used two workflows based on either pressure cycling technology (PCT) or ultrasonication (US) in combination with data independent acquisition (DIA) based mass spectrometry (MS) analysis to quantitatively assess the proteomes of articular cartilage and subchondral bone.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Equus Caballus (horse)

TISSUE(S): Articular Cartilage, Bone, Synovial Tissue

SUBMITTER: Louise Bundgaard

LAB HEAD: Louise Bundgaard

PROVIDER: PXD025882 | Pride | 2022-10-12

REPOSITORIES: pride

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Effective protein extraction combined with data independent acquisition analysis reveals a comprehensive and quantifiable insight into the proteomes of articular cartilage and subchondral bone.

Bundgaard L L Åhrman E E Malmström J J Auf dem Keller U U Walters M M Jacobsen S S

Osteoarthritis and cartilage 20210920 1

<h4>Objective</h4>The objectives of this study was to establish a sensitive and reproducible method to map the cartilage and subchondral bone proteomes in quantitative terms, and mine the proteomes for proteins of particular interest in the pathogenesis of osteoarthritis (OA). The horse was used as a model animal.<h4>Design</h4>Protein was extracted from articular cartilage and subchondral bone samples from three horses in triplicate by pressure cycling technology or ultrasonication. Digested pr ...[more]

PMID: 34547431

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Project description:Objective : To study molecular changes in the articular cartilage and subchondral bone of the tibial plateau from mice deficient in frizzled related protein (Frzb) compared to wild-type mice by transcriptome analysis. Methods : Gene-expression analysis of the articular cartilage and subchondral bone of 3 wild-type and 3 Frzb-/- mice was performed by microarray. Pathway analysis of differentially expressed genes between 3 wild-type and 2 Frzb-/- samples was explored with PANTHER, DAVID and GSEA bioinformatics tools. Activation of the WNT pathway was analyzed using western blot. The effects of Frzb gain and loss of function on chondrogenesis and cell proliferation was examined using ATDC5 micromasses and mouse ribcage chondrocytes. Results : Extracellular matrix-associated integrin and cadherin pathways, as well as WNT pathway genes were upregulated in Frzb-/- samples. Several WNT receptors, target genes, and other antagonists were upregulated, but no difference in active β-catenin was found. Analysis of ATDC5 cell micromasses overexpressing FRZB indicated an upregulation of aggrecan and Col2a1, and downregulation of molecules related to damage and repair in cartilage, Col3a1 and Col5a1. Silencing of Frzb resulted in downregulation of aggrecan and Col2a1. Pathways associated with cell cycle were downregulated. Ribcage chondrocytes derived from Frzb-/- mice showed decreased proliferation compared to wild-type cells. Conclusions : Our analysis provides evidence for tight regulation of WNT signaling, shifts in extracellular matrix components and effects on cell proliferation and differentiation in the articular cartilage - subchondral bone unit in Frzb-/- mice. These data further support an important role for FRZB in joint homeostasis and highlight the complex biology of WNT signaling in the joint. Gene-expression analysis of the articular cartilage and subchondral bone of 3 wild-type and 3 Frzb-/- mice was performed by microarray. Pathway analysis of differentially expressed genes between 3 wild-type and 2 Frzb-/- samples was explored with PANTHER, DAVID and GSEA bioinformatics tools.

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	20181105_FJ_LB_CAR4.raw	Raw
	20181105_FJ_LB_CAR6.raw	Raw
	20181105_FJ_LB_SB7.raw	Raw
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