Supplementary material Ann Rheum Dis Vanarsa K, et al. Ann Rheum Dis 2020;0:1–13. doi: 10.1136/annrheumdis-2019-216312 Supplementary material Ann Rheum Dis Vanarsa et al, Supplementary Table S2 Supplementary Table S2: Patient cohort used for the ELISA validation of urine biomarker candidates Variable All SLE Inactive Active Non-Renal Active Renal (N=62) (N=17) (N=16) (N=29) Age (years) 32.9±9.5 32.8±7.5 42.1±11.3 27.9±4.5 Gender Female, n(%) 62 (100.0%) 17 (100.0%) 16 (100.0%) 29 (100.0%) Ethnicity African-American 58 (93.5%) 16 (94.1%) 13 (81.3%) 29 (100.0%) Caucasian 4 (6.5%) 1 (5.9%) 3 (18.7%) 0 (0.0%) Clinical Assessment SLEDAI 5.9±4.2 0.4±0.8 7.7±3.0 8.1±3.0 rSLEDAI 2.4±2.9 0±0 0±0 5.1±2.1 PGA 1.3±0.8 0.6±0.7 0.8±0.4 2.0±0.6 Laboratory Metrics WBC (X109/L) 6.9±3.3 7.1±2.7 4.3±1.4 8.3±3.6 HGB (g/dl) 11.0±1.5 11.3±1.5 11.3±1.1 10.7±1.7 Platelets (X109/L) 284.0±95.8 235.9±78.9 263.5±62.1 323.2±106.0 C3 (mg/dl) 95.3±34.7 116.5±31.0 87.8±37.5 88.3±31.1 C4 (mg/dl) 18.7±10.2 22.0±8.7 14.9±8.9 19.1±11.1 ESR (mm/h) 43.9±29.2 34.1±26.3 47.9±22.0 47.2±33.5 UrPrCrRatio (mg/mg) 1.2±1.9 0.2±0.1 0.1±0.1 2.5±2.4 Serum Creatinine (mg/dl) 0.9±0.4 0.9±0.5 0.8±0.2 1.0±0.4 Anti-DNA, n(%) 32 (51.6%) 2 (11.8%) 11 (68.8%) 19 (65.5%) Medications Prednisone, n(%) 38 (61.3%) 9 (52.9%) 7 (43.8%) 22 (75.9%) Plaquenil, n(%) 51 (82.3%) 13 (76.5%) 13 (81.3%) 25 (86.2%) Mycophenolate Mofetil, n(%) 32 (51.6%) 8 (47.1%) 9 (56.3%) 15 (51.7%) Tacrolimus, n(%) 1 (1.6%) 0 (0.0%) 1 (6.3%) 0 (0.0%) Azathioprine, n(%) 7 (11.3%) 1 (5.9%) 2 (12.5%) 4 (13.8%) Cyclophosphamide, n(%) 2 (3.2%) 0 (0.0%) 0 (0.0%) 2 (6.9%) Methotrexate, n (%) 2 (3.2%) 0 (0.0%) 1 (6.3%) 1 (3.4%) Abbreviations used: PGA, physician global assessment; UrPrCr ratio, urine protein to creatinine ratio. Vanarsa K, et al. Ann Rheum Dis 2020;0:1–13. doi: 10.1136/annrheumdis-2019-216312 Supplementary material Ann Rheum Dis Supplementary Table S3: Well documented biomarker proteins rediscovered by the screen Molecule FC SLE/HC p value (t test) Supplementary References Albumin 2.0 0.075 S3, S4 ALCAM 1797.9 0.072 S47, S48 Angiostatin 160.9 0.105 S29 Calreticulin 4.6 0.1 S44 CD163 N/A* 0.08 S10, S11 CD30 20.5 0.008 S51 CXCL16 15.9 0.008 S20, S21 Cystatin S 3.0 0.185 Cystatin SA 1.7 0.529 Cystatin SN 0.4 0.313 E-Selectin 3.9 0.002 S37 Ferritin 187.0 0.001 S49, S50 FOLR1 1.4 0.517 ICAM-1 6.8 0.019 S1, S2 IL-12p40 N/A* 0.3 S35, S36 IL-15 4.1 0.47 S13, S14 IL-6 0.1 0.126 S5, S6 IP-10 43.1 0.013 S22, S23 I-TAC 2.5 0.137 Leptin 3.1 0.024 S12 Lipocalin-1 3.1 0.055 Lipocalin-2 3.2 0.012 S28 L-Selectin 16.7 0.013 MCP-1 7.8 0.089 S7 MCSF 1.5 0.344 S15, S16 MCSF R 51.9 0.008 MIP-3a 49.8 0.13 S27 MIP-3b 8.8 0.04 S45 OPG 52.0 0.002 S33, S34 OPN 7.2 0.004 S18, S19 PF4 43.1 0.109 S40, S41 RAGE 10.7 0.1 S26 RANTES 16.7 0.01 S42, S43 Resistin 8.1 0.015 S46 SDF-1a 9.1 0.249 S32 TFPI 397.8 0.027 S8, S9 TNF RI 3.7 0.038 S17 TNF RII 3.2 0.025 TREM-1 21.7 0.099 S38, S39 TWEAK 4.4 0.105 S30, S31 TWEAK R 1.5 0.743 uPA 3.7 0.004 VCAM-1 49.0 0.068 S24, S25 N/A, not available. * indicates FC was not calculated due to average of healthy control is 0. Vanarsa K, et al. Ann Rheum Dis 2020;0:1–13. doi: 10.1136/annrheumdis-2019-216312 Supplementary material Ann Rheum Dis Supplementary References: S1 Guan J, Wang G, Tam L-S, et al. Urinary sediment ICAM-1 level in lupus nephritis. Lupus 2012;21:1190–5. doi:10.1177/0961203312451334 S2 Sabry A, Sheashaa H, El-Husseini A, et al. Intercellular adhesion molecules in systemic lupus erythematosus patients with lupus nephritis. Clin Rheumatol 2007;26:1819–23. doi:10.1007/s10067-007-0580-7 S3 Ding J, Zheng Z, Li X, et al. Urinary Albumin Levels are Independently Associated with Renal Lesion Severity in Patients with Lupus Nephritis and Little or No Proteinuria. Med Sci Monit 2017;23:631–9. doi:10.12659/MSM.899973 S4 Yip J, Aghdassi E, Su J, et al. Serum albumin as a marker for disease activity in patients with systemic lupus erythematosus. J Rheumatol 2010;37:1667–72. doi:10.3899/jrheum.091028 S5 Jacob N, Stohl W. Cytokine disturbances in systemic lupus erythematosus. Arthritis Research & Therapy 2011;13:228. doi:10.1186/ar3349 S6 Ripley B, Goncalves B, Isenberg D, et al. Raised levels of interleukin 6 in systemic lupus erythematosus correlate with anaemia. Ann Rheum Dis 2005;64:849–53. doi:10.1136/ard.2004.022681 S7 Živković V, Cvetković T, Mitić B, et al. Monocyte chemoattractant protein-1 as a marker of systemic lupus erythematosus: an observational study. Rheumatol Int 2018;38:1003–8. doi:10.1007/s00296- 017-3888-x S8 Qin L, Stanley S, Ding H, et al. Urinary pro-thrombotic, anti-thrombotic, and fibrinolytic molecules as biomarkers of lupus nephritis. Arthritis Res Ther 2019;21:176. doi:10.1186/s13075-019-1959-y S9 Adams MJ, Palatinus AA, Harvey AM, et al. Impaired control of the tissue factor pathway of blood coagulation in systemic lupus erythematosus. Lupus 2011;20:1474–83. doi:10.1177/0961203311418267 S10 Endo N, Tsuboi N, Furuhashi K, et al. Urinary soluble CD163 level reflects glomerular inflammation in human lupus nephritis. Nephrol Dial Transplant 2016;31:2023–33. doi:10.1093/ndt/gfw214 S11 Nakayama W, Jinnin M, Makino K, et al. CD163 expression is increased in the involved skin and sera of patients with systemic lupus erythematosus. Eur J Dermatol 2012;22:512–7. doi:10.1684/ejd.2012.1756 S12 Hutcheson J, Ye Y, Han J, et al. Resistin as a potential marker of renal disease in lupus nephritis. Clin Exp Immunol 2015;179:435–43. doi:10.1111/cei.12473 S13 Stanley S, Mok CC, Vanarsa K, et al. Identification of Low-Abundance Urinary Biomarkers in Lupus Nephritis Using Electrochemiluminescence Immunoassays. Arthritis & Rheumatology (Hoboken, NJ) 2019;71:744–55. doi:10.1002/art.40813 S14 Liu C-C, Kao AH, Manzi S, et al. Biomarkers in systemic lupus erythematosus: challenges and prospects for the future. Ther Adv Musculoskelet Dis 2013;5:210–33. doi:10.1177/1759720X13485503 S15 Tian S, Li J, Wang L, et al. Urinary levels of RANTES and M-CSF are predictors of lupus nephritis flare. Inflamm Res 2007;56:304–10. doi:10.1007/s00011-007-6147-x Vanarsa K, et al. Ann Rheum Dis 2020;0:1–13. doi: 10.1136/annrheumdis-2019-216312 Supplementary material Ann Rheum Dis S16 Ramirez GA, Blasi M, Sciorati C, et al. Plasma levels of M-CSF are increased in ANCA-associated vasculitides with active nephritis. Results Immunol 2015;5:33–6. doi:10.1016/j.rinim.2015.10.002 S17 Adhya Z, El Anbari M, Anwar S, et al. Soluble TNF-R1, VEGF and other cytokines as markers of disease activity in systemic lupus erythematosus and lupus nephritis. Lupus 2019;28:713–21. doi:10.1177/0961203319845487 S18 Kitagori K, Yoshifuji H, Oku T, et al. Cleaved Form of Osteopontin in Urine as a Clinical Marker of Lupus Nephritis. PLoS One 2016;11. doi:10.1371/journal.pone.0167141 S19 Pasha HF, Tantawy EA, Youssef MA. Osteopontin and interleukin-17A genes polymorphisms in Egyptian systemic lupus erythematosus patients: A relation to disease activity and severity. Gene 2019;702:107–13. doi:10.1016/j.gene.2019.02.100 S20 Reyes-Thomas J, Blanco I, Putterman C. Urinary Biomarkers in Lupus Nephritis. Clin Rev Allergy Immunol 2011;40:138–50. doi:10.1007/s12016-010-8197-z S21 Qin M, Guo Y, Jiang L, et al. Elevated levels of serum sCXCL16 in systemic lupus erythematosus; potential involvement in cutaneous and renal manifestations. Clin Rheumatol 2014;33:1595–601. doi:10.1007/s10067-014-2741-9 S22 Zeid MMH, Baddour NM, El-Neily DAE-M, et al. Study of urinary interferon gamma-induced protein 10 (IP-10) and urinary soluble CD 25 (sCD25) as markers of lupus nephritis and their relation to histological class.