Dataset Information

Proteolytic information flows from human knee osteoarthritic cartilage to synovial fluid and defines the roles of key drug targets -1

ABSTRACT: Objectives: Proteolytic cartilage loss is a major mechanism of osteoarthritis (OA) pathogenesis but few joint breakdown biomarkers are currently available. We sought to determine the overlap of proteolytic peptides in OA cartilage and synovial fluid on a proteome-wide scale with the goals of increasing the biomarker repertoire and for attribution to individual proteases. Design: Proteins isolated from matched human OA cartilage and synovial fluid from 5 knees were analyzed by N-terminomics using Terminal Amine Isotopic Labeling of Substrates (TAILS), comprising labeling and enrichment of protein N-termini, high-resolution mass spectrometry and bioinformatics. Cartilage was digested with ADAMTS5, MMP13 and CMA1, and TAILS was used to identify their specific activities. Results: Analysis of matched knee OA cartilage and synovial fluid degradomes consistently demonstrated cartilage breakdown products in synovial fluid. Of over 20,000 cleaved peptides in the OA cartilage and synovial fluid degradomes, 677, originating from 153 proteins, were present in all samples, the majority arising from cartilage. ADAMTS5, MMP13 and CMA1 digestion of cartilage identified numerous cleavage sites for each protease, distinct cleavage site preferences, and peptides arising from the activities of these proteases in synovial fluid. Conclusions: Cartilage ECM-derived proteolytic fragments are consistently present in synovial fluid, many attributable to ADAMTS5, MMP13 and CMA1 activity, which showed little overlap with each other or the previously determined HtrA1activity profile. The present work increases the proteolytic biomarker space for OA investigation, showing that diverse proteases contribute to cartilage destruction, and that their individual contributions can be determined using multiple protease-specific biomarkers.

INSTRUMENT(S): Bruker Daltonics timsTOF series

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cartilage

DISEASE(S): Osteoarthritis

SUBMITTER: Sumit Bhutada

LAB HEAD: Suneel Apte

PROVIDER: PXD048053 | Pride | 2025-02-04

REPOSITORIES: pride

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Dataset's files

Source:

			Action	DRS
	CMA_Human_cartilage_TAILS1_Fr1.d.zip	Other
	CMA_Human_cartilage_TAILS1_Fr1.pepXML	Pepxml
	CMA_Human_cartilage_TAILS1_Fr2.d.zip	Other
	CMA_Human_cartilage_TAILS1_Fr2.pepXML	Pepxml
	CMA_Human_cartilage_TAILS1_Fr3.d.zip	Other

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Publications

Degradomics defines proteolysis information flow from human knee osteoarthritis cartilage to matched synovial fluid and the contributions of secreted proteases ADAMTS5, MMP13 and CMA1 to articular cartilage breakdown.

Bhutada Sumit S Hoyle Anna A Piuzzi Nicolas S NS Apte Suneel S SS

Osteoarthritis and cartilage 20240916 1

<h4>Objectives</h4>Proteolytic cartilage extracellular matrix breakdown is a major mechanism of articular cartilage loss in osteoarthritis (OA) pathogenesis. We sought to determine the overlap of proteolytic peptides in matched knee OA cartilage and synovial fluid on a proteome-wide scale to increase the prospective biomarker repertoire and to attribute proteolytic cleavages to specific secreted proteases.<h4>Design</h4>Matched human knee OA cartilage and synovial fluid (n = 5) were analyzed by ...[more]

PMID: 39293776

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