Synovial fluid biomarkers associated with osteoarthritis severity reflect macrophage and neutrophil related inflammation

Collin A Haraden, Janet L Huebner, Ming-Feng Hsueh, Yi-Ju Li, Virginia Byers Kraus, Collin A Haraden, Janet L Huebner, Ming-Feng Hsueh, Yi-Ju Li, Virginia Byers Kraus

Abstract

Background: To identify a synovial fluid (SF) biomarker profile characteristic of individuals with an inflammatory osteoarthritis (OA) endotype.

Methods: A total of 48 knees (of 25 participants) were characterized for an extensive array of SF biomarkers quantified by Rules Based Medicine using the high-sensitivity multiplex immunoassay, Myriad Human InflammationMAP® 1.0, which included 47 different cytokines, chemokines, and growth factors related to inflammation. Multivariable regression with generalized estimating equations (GEE) and false discovery rate (FDR) correction was used to assess associations of SF RBM biomarkers with etarfolatide imaging scores reflecting synovial inflammation; radiographic knee OA severity (based on Kellgren-Lawrence (KL) grade, joint space narrowing, and osteophyte scores); knee joint symptoms; and SF biomarkers associated with activated macrophages and knee OA progression including CD14 and CD163 (shed by activated macrophages) and elastase (shed by activated neutrophils).

Results: Significant associations of SF biomarkers meeting FDR < 0.05 included soluble (s)VCAM-1 and MMP-3 with synovial inflammation (FDR-adjusted p = 0.025 and 1.06 × 10-7); sVCAM-1, sICAM-1, TIMP-1, and VEGF with radiographic OA severity (p = 1.85 × 10-5 to 3.97 × 10-4); and VEGF, MMP-3, TIMP-1, sICAM-1, sVCAM-1, and MCP-1 with OA symptoms (p = 2.72 × 10-5 to 0.050). All these SF biomarkers were highly correlated with macrophage markers CD163 and CD14 in SF (r = 0.43 to 0.90, FDR < 0.05); all but MCP-1 were also highly correlated with neutrophil elastase in SF (r = 0.62 to 0.89, FDR < 0.05).

Conclusions: A subset of six SF biomarkers was related to synovial inflammation in OA, as well as radiographic and symptom severity. These six OA-related SF biomarkers were specifically linked to indicators of activated macrophages and neutrophils. These results attest to an inflammatory OA endotype that may serve as the basis for therapeutic targeting of a subset of individuals at high risk for knee OA progression.

Trial registration: Written informed consent was received from participants prior to inclusion in the study; the study was registered at ClinicalTrials.gov ( NCT01237405 ) on November 9, 2010, prior to enrollment of the first participant.

Keywords: Biomarker; Inflammation; Joint pain; Knee; Macrophage; Neutrophil; Osteoarthritis; Radiograph; Severity.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
STRING plot showing functional biomarker interrelationships. Depicted relationships represent biomarkers with significant associations with OA inflammation, radiographic OA, and OA symptoms analyzed using the STRING v10.5 database. The colors of the spheres correspond to the biological processes in which a particular biomarker is involved. Sphere color key: leukocyte migration (red), extracellular matrix organization (blue), and inflammatory response (green). Joining string color key (corresponding to origin of data used for STRING database): curated databases (blue), experimentally determined (pink), textmining (yellow), and co-expression (black). Abbreviations: CD, cluster of differentiation 14; CD163, cluster of differentiation 163; CCL2 (also referred to as MCP-1 or monocyte chemoattractant protein 1), C-C chemokine motif 2 vascular cell adhesion molecule 1; ELANE, neutrophil elastase; ICAM-1, intracellular adhesion molecule 1; MMP-3, matrix metalloproteinase-3; TIMP-1, tissue inhibitor of metallopeptidase inhibitor 1; VEGF, vascular endothelial growth factor
Fig. 2
Fig. 2
Inflammatory OA endotype based on synovial fluid biomarkers. This plot conceptualizes our data in the context of the current understanding of inflammation in OA. Abbreviations: OA, osteoarthritis; DAMPs, disease-associated molecular patterns; VCAM-1, vascular cell adhesion molecule 1; MMP-3, matrix metalloproteinase-3; ICAM-1, intracellular adhesion molecule 1; TLR, toll-like receptor; TIMP-1, tissue inhibitor of metallopeptidase inhibitor 1; VEGF, vascular endothelial growth factor; MCP-1, monocyte chemoattractant protein 1; KL, Kellgren-Lawrence; JSN, joint space narrowing; OST, osteophyte; NHANES, national health and nutrition examination survey measure of OA symptoms (pain, aching, stiffness)

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