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Soluble Macrophage Biomarkers Indicate Inflammatory Phenotypes in Patients with Knee Osteoarthritis

Hikmat N. Daghestani, PhD1,2; Carl F. Pieper, DPH3; and Virginia B. Kraus, MD, PhD1,2* Affiliations: 3 Duke Molecular Physiology Institute1, Departments of Medicine2 and Biostatistics and

Bioinformatics3, Duke University School of Medicine, Durham, NC

*Address correspondence to Virginia B. Kraus, MD, PhD, Duke Molecular Physiology

Institute, PO Box 104775, Carmichael Building, 300 N. Duke St., Durham, NC 27701.

Tel: 919-681-6652; Fax: 919-684-8907; E-mail: vbk@duke.edu.

Keywords: Inflammation; biomarker; knee osteoarthritis; macrophages Financial Support: EC20 scan samples were obtained with support of a grant from Lilly

USA, Inc. This work was supported by P01 AR050245 (VBK), and NIH 5T32AI007217-30

(HND).

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Supplementary Tables: Supplementary Table 1. Associations of synovial fluid CD163 and CD14 concentrations with osteoarthritis (OA) phenotypes with end-stage OA (KL4) knee samples excluded.

OA Phenotype

Synovial Fluid CD163

Synovial Fluid CD14

β* 95% CI P-value β* 95% CI P-value

Activated macrophages in joint capsulea

0.750 0.152, 1.348 0.014 0.750 -0.007, 1.508 0.052

Activated macrophages in synoviuma

1.644 0.937, 2.352 <0.0001 1.718 0.817, 2.620 0.0002

OST severitya 2.401 0.515, 4.288 0.013 2.474 0.357, 4.592 0.022

OST severityb 0.052 -0.115, 0.219 0.541 0.574 0.269, 0.880 0.0002

JSN severitya 0.653 0.356, 0.767 <0.0001 0.920 0.616, 1.225 <0.0001

JSN severityb 0.027 -0.001, 0.055 0.056 0.074 -0.048, 0.196 0.234

OST progressionb 0.064 0.024, 0.104 0.002 0.061 -0.033, 0.155 0.206

JSN progressionb 0.065 -0.014, 0.027 0.537 0.006 -0.059, 0.072 0.848 a. Etarfolatide (EC20) scan cohort, b. Prediction of Osteoarthritis Progression (POP) cohort. *Adjusted for age, gender, and body mass index. β, parameter estimate (based on CD163 and CD14 in µg/ml); 95% CI, 95% confidence interval; OST, osteophyte; JSN, joint space narrowing.

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Supplementary Table 2. Association of serum CD163 and plasma CD14 concentrations with osteoarthritis (OA) phenotypes with end-stage OA (KL4) knee samples excluded.

OA Phenotype

Serum CD163

Plasma CD14

β* 95% CI P-value β* 95% CI P-value Activated

macrophages in joint capsulea

1.724 0.504, 2.944 0.006 0.874 0.352, 1.395 0.001

Activated macrophages in synoviuma

3.153 1.694, 4.612 < 0.0001 0.916 -0.221, 2.052 0.114

OST severitya 1.405 -3.459, 6.270 0.571 -0.173 -2.150, 1.803 0.864

OST severityb -1.001 -1.768, -0.233 0.011 0.170 -0.424, 0.764 0.575

JSN severitya 0.542 -0.077, 1.160 0.086 0.065 -0.526, 0.656 0.829

JSN severityb 0.069 -0.243, 0.381 0.666 -0.007 -0.219, 0.205 0.950

OST progressionb -0.102 -0.366, 0.162 0.450 0.103 -0.123, 0328 0.373

JSN progressionb -0.010 -0.132, 0.112 0.867 0.108 -0.012, 0.228 0.078 a. Etarfolatide (EC20) scan cohort, b. Prediction of Osteoarthritis Progression (POP) cohort. *Adjusted for age, gender, and body mass index. β, parameter estimate (based on CD163 and CD14 in µg/ml); 95% CI, 95% confidence interval; TF, tibiofemoral; OST, osteophyte; JSN, joint space narrowing.

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Supplementary Table 3. Association of CD163 and CD14 with joint symptoms with end-stage osteoarthritis (KL4) knee samples excluded.

Sample Type

CD163

CD14

β* 95% CI P-value β* 95% CI P-value

Synovial fluida 0.179 -0.213, 0.571 0.370 0.442 0.017, 0.866 0.042

Synovial fluidb -0.051 -0.126, 0.025 0.188 0.038 -0.087, 0.162 0.5525

Serum/Plasmaa 1.106 0.528, 1.683 0.0002 0.479 0.215, 0.744 0.0004

Serum/Plasmab 0.317 -0.107, 0.380 0.270 0.134 -0.125, 0.393 0.3118

a. Etarfolatide (EC20) scan cohort, b. Prediction of Osteoarthritis Progression (POP) cohort. *Adjusted for age, gender, and body mass index. β, parameter estimate (based on CD163 and CD14 in µg/ml); 95% CI, 95% confidence interval.

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Supplementary Table 4. The combinatorial effect of CD163 and CD14 with osteoarthritis phenotypes assessed by quasi-likelihood information criterion (QICu) in the Etarfolatide (EC20) scan cohort.

OST

severity JSN

severity

Activated macrophages

in joint capsule

Activated macrophages in synovium

Joint symptoms

Plasma CD14 44 44 44 44 44

Serum CD163 44 44 44 44 44

Plasma CD14 + Serum CD163 45 45 45 45 45

SF CD14 38 38 38 38 38

SF CD163 42 42 42 42 42

SF CD14 + SF CD163 39 39 39 39 39

OST, osteophyte; JSN, joint space narrowing; SF, synovial fluid. A lower QICu value indicates a more suitable model,

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Supplementary Table 5. The combinatorial effect of CD163 and CD14 with osteoarthritis phenotypes assessed by quasi-likelihood information criterion (QICu) in the Prediction of Osteoarthritis Progression (POP) cohort.

OST

severity JSN

severity OST

progression JSN

progression Joint

symptoms

Plasma CD14 311 312 249 249 290

Serum CD163 309 310 247 247 288

Plasma CD14 + Serum CD163 310 311 248 248 289

SF CD14 175 175 135 135 161

SF CD163 174 174 131 131 156

SF CD14 + SF CD163 159 159 122 122 145

OST, osteophyte; JSN, joint space narrowing; SF, synovial fluid. A lower QICu value indicates a more suitable model,