Supplementary Table 2: Non-proteomic studies to discover biomarkers of bladder cancer
Biomarker(s) ‡: / Samples† / Study design and Diagnostic/Prognostic Utility‡ / Reference(s):MMP1 / n = 131 bladder tumour samples, n = 5 prostate cancer, n = 33 benign lower urinary tract infections, n = 36 healthy volunteer. / ELISA-based study:
Patients with detectable levels of urinary MMP1 had higher disease progression rates (P = 0.04). / (1)
MMP2 / n = 82 bladder carcinoma patients, n = 28 control patients. / ELISA and gelatin zymography, Detects early tumour progression and elevated MMPs levels correlate with tumour stage. / (2)
n = 44 controls, n = 14 cystitis patients, n = 43 stage Ta-T1, n = 18 stage T2, n = 10 stage T3-T4, / Gelatine zymography, Elevated in bladder cancer and correlates with disease stage.Sensitivity to detect bladder cancer = 0.51. / (3)
MMP9 / n = 82 bladder cancer patients (43 = pTa, 28 = pT1, 6 = pT2, 4 = pT3) and 28 controls. / ELISA and gelatin zymography.Elevated MMP9 detects bladder cancer but with specificity similar to urine cytology / (2)
n = 44 control urine samples, n = 14 cystitis patients, n = 43 stage Ta-T1, n = 18 stage T2, n = 10 stage T3-T4, / Gelatine zymography, Sensitivity to detect bladder cancer = 0.31. MMP9 levels observed to be elevated particularly within patients with muscle invasive bladder cancer. / (3)
TIMP2 / n = 154 bladder cancer patients, 60 benign urological condition patients, 30 healthy volunteers. / ELISA/zymography based study: Urinary sediments used for cytology and within the supernatant MMP2 : TIMP2 and MMP9 : TIMP2 ratios inversely correlated with tumour stage and grade / (4)
γ-synuclein, catechol-o-methyltransferase and reticulin / n = 112 bladder cancer donors and n = 230 control donors. / Quantitative-WB: With optimal cut-off panel is diagnostic of bladder cancer with 76.8% sensitivity and 77.4% specificity. / (5)
EGF / n = 54 bladder cancer donors (16 grade 1Ta, 38 grade 3 or T2> lesions) and n = 66 pathological or normal control samples without bladder cancer. / Radioimmunoassay. EGF is decreased in patients with bladder cancer (p < 0.0001). / (6)
DEK-Protein / n = 38 bladder cancer tissue sections, n = 23 normal urine samples and n = 19 bladder cancer urine samples. / Quantitative WB and ELISA:DEK protein expressed in 33 out of 38 bladder tumours-not in adjacent normal healthy tissues), and within 23 normal urine samples and 16 out of 19 bladder cancer urine samples. 84% of transitional cell carcinoma urine samples were tested positive for the presence of DEK-protein. / (7)
Cytokeratin-based tests. / n=44 patients with bladder cancer as assessed by cytoscopy and n=29 patients without bladder cancer by cytoscopy. / ELISA test for cytokeratin 8/18 and UBC ELISA test). Combination of the UBC antigen test with the proportion of cells showing a DNA content >2n increased the sensitivity of the UBC test to 89%.. / (8)
n = 111 active bladder cancer patients, n = 76 disease-free controls. / CYFRA21.1 test. At specificity of 95% cut-off values for tests were: CYFRA21.1 = 5.4 micrograms/L, 15.4 micrograms/L for bladder cancer antigen,760.8 units/L for tissue polypeptide antigen and 14.6 U/ml for NMP22. At these cut-offs the sensitivity of the CYFRA21.1 test for detection bladder cancer was 83.8%. / (9)
n = 152 patients with either haematurias or irritative voiding symptoms, n = 107 patients under bladder cancer surveillance, n = 46 non-bladder cancer urinary tract pathologies, n = 20 healthy donors. / Immunoradiometric Assay. At 4.9 microgrammes/L CK19 fragments detect bladder cancer with sensitivity = 79.3% and specificity = 88.6%. / (10)
Mucin-like antigens (M344, 19A211 and LDQ10 against transitional cell carcinoma in exfoliated urothelial cells ) and glycosylated carcinoembryonic antigen / n = 150 donors post bladder cancer resection, n = 114 donors with bladder-cancer symptoms but not diagnosed. / Fluorescent antibody-based detection-ImmunoCyt test: Multiple-antigen test detects bladder cancer with 86.1% sensitivity and 79.4% specificity. / (11)
Hyaluronic acid and hyaluronidase / n=30 bladder cancer patient urines (14 stage pTa, 9 stage pT1, 5 stage pT2 and 2 with carcinoma in situ transitional cell carcinoma) and n=64 controls (55 with a history of bladder cancer but no cytoscopic evidence of bladder cancer at the time of sampling and 9 benign prostatic hyperplasia samples). / HA-HAase test-type test. Increased hyaluronidase activity detects bladder cancer with 83.3% sensitivity and 78.1% specificity. / (12)
Soluble Fas receptor isoforms / n = 188 patient urine samples including 31 non-cancer urologic controls and 10 healthy controls. / ELISA.ROC AUC for sFAS receptor = 0.757. / (13)
Nuclear Matrix Protein-22 (NMP22) / n = 43 newly diagnosed bladder cancer patients, n = 58 recurrent bladder cancer, n = 77 remission patients and n = 25 healthy controls. / ELISA based study (Bladder Chek). NMP22 detects bladder cancer with sensitivity of 88% (newly diagnosed) and 57% (recurrent), mean specificity = 67%. / (14)
HSP60 and IL-13 / Discovery set: n = 18 healthy donors, n = 20 haematuria patients, n = 50 non-muscle-invasive bladder cancer, n = 18 muscle invasive bladder cancer. Validation set = n=19 control patients, n=21 bladder cancer. / ELISA based study. ROC AUC for IL-13 = 0.93 for prediction of bladder cancer. Combination of HSP60 and IL-13 predicted bladder cancer with positive predictive value = 74% and negative predictive value = 76%. / (15)
Telomerase / n = 197 patients with urinary tract symptoms and n=318 with diagnosis of bladder cancer. / Enzymatic Assay within urine. At 50 units cut-off for bladder cancer diagnosis, sensitivity = 87%, specificity = 70%. / (16)
Uroplakin-3A (UPK3A) / Urine samples, n = 32 healthy controls, n = 44 benign urological conditions, n = 122 bladder cancer donors. / ELISA-based study. UPK3A elevated in bladder cancer. At cut-off of 0.0685 absorbance units, sensitivity and specificity for UPK3A test = 83%. ROC AUC = 0.907. / (17)
Mini-Chromosome Maintenance-5 (MCM5) protein. / Total study group, n = 1677 consecutive patients. / Immunofluorimetric assay. Increased MCM5-protein predictive of bladder cancer. Mini-chromosome maintenance-5 protein test detects bladder cancer with 69% specificity and 93% negative predictive value. ROC AUC with optimal MCM5 cut-off for diagnosis of bladder cancer was 0.75. / (18)
Clusterin / n = 68 bladder cancer donors, n = 61 benign urological conditions. / ELISA-based study. Clusterin elevated in bladder cancer and for detection of bladder cancer sensitivity and specificity = 87.1% and 96.7% respectively. / (19)
†All samples are human and urine unless otherwise specified; n numbers refer to number with condition unless otherwise stated,
‡Abbreviations used for proteomic techniques are defined within the text and list of common abbreviations.
1.Durkan GC, Nutt JE, Rajjayabun PH, Neal DE, Lunec J, Mellon JK. Prognostic significance of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 in voided urine samples from patients with transitional cell carcinoma of the bladder. Clin Cancer Res. 2001 Nov;7(11):3450-6.
2.Sier CF, Casetta G, Verheijen JH, Tizzani A, Agape V, Kos J, et al. Enhanced urinary gelatinase activities (matrix metalloproteinases 2 and 9) are associated with early-stage bladder carcinoma: a comparison with clinically used tumor markers. Clin Cancer Res. 2000 Jun;6(6):2333-40.
3.Gerhards S, Jung K, Koenig F, Daniltchenko D, Hauptmann S, Schnorr D, et al. Excretion of matrix metalloproteinases 2 and 9 in urine is associated with a high stage and grade of bladder carcinoma. Urology. 2001 Apr;57(4):675-9.
4.Eissa S, Ali-Labib R, Swellam M, Bassiony M, Tash F, El-Zayat TM. Noninvasive diagnosis of bladder cancer by detection of matrix metalloproteinases (MMP-2 and MMP-9) and their inhibitor (TIMP-2) in urine. Eur Urol. 2007 Nov;52(5):1388-96.
5.Iwaki H, Kageyama S, Isono T, Wakabayashi Y, Okada Y, Yoshimura K, et al. Diagnostic potential in bladder cancer of a panel of tumor markers (calreticulin, gamma -synuclein, and catechol-o-methyltransferase) identified by proteomic analysis. Cancer Sci. 2004 Dec;95(12):955-61.
6.Messing EM, Murphy-Brooks N. Recovery of epidermal growth factor in voided urine of patients with bladder cancer. Urology. 1994 Oct;44(4):502-6.
7.Datta A, Adelson ME, Mogilevkin Y, Mordechai E, Sidi AA, Trama JP. Oncoprotein DEK as a tissue and urinary biomarker for bladder cancer. BMC Cancer. 2011;11:234.
8.Sanchez-Carbayo M, Ciudad J, Urrutia M, Navajo JA, Orfao A. Diagnostic performance of the urinary bladder carcinoma antigen ELISA test and multiparametric DNA/cytokeratin flow cytometry in urine voided samples from patients with bladder carcinoma. Cancer. 2001 Dec 1;92(11):2811-9.
9.Sanchez-Carbayo M, Herrero E, Megias J, Mira A, Soria F. Comparative sensitivity of urinary CYFRA 21-1, urinary bladder cancer antigen, tissue polypeptide antigen, tissue polypeptide antigen and NMP22 to detect bladder cancer. J Urol. 1999 Dec;162(6):1951-6.
10.Nisman B, Barak V, Shapiro A, Golijanin D, Peretz T, Pode D. Evaluation of urine CYFRA 21-1 for the detection of primary and recurrent bladder carcinoma. Cancer. 2002 Jun 1;94(11):2914-22.
11.Mian C, Pycha A, Wiener H, Haitel A, Lodde M, Marberger M. Immunocyt: a new tool for detecting transitional cell cancer of the urinary tract. J Urol. 1999 May;161(5):1486-9.
12.Hautmann S, Toma M, Lorenzo Gomez MF, Friedrich MG, Jaekel T, Michl U, et al. Immunocyt and the HA-HAase urine tests for the detection of bladder cancer: a side-by-side comparison. Eur Urol. 2004 Oct;46(4):466-71.
13.Svatek RS, Herman MP, Lotan Y, Casella R, Hsieh JT, Sagalowsky AI, et al. Soluble Fas--a promising novel urinary marker for the detection of recurrent superficial bladder cancer. Cancer. 2006 Apr 15;106(8):1701-7.
14.Kehinde EO, Al-Mulla F, Kapila K, Anim JT. Comparison of the sensitivity and specificity of urine cytology, urinary nuclear matrix protein-22 and multitarget fluorescence in situ hybridization assay in the detection of bladder cancer. Scand J Urol Nephrol. 2011 Mar;45(2):113-21.
15.Margel D, Pevsner-Fischer M, Baniel J, Yossepowitch O, Cohen IR. Stress proteins and cytokines are urinary biomarkers for diagnosis and staging of bladder cancer. Eur Urol. 2011 Jan;59(1):113-9.
16.Casadio V, Bravaccini S, Gunelli R, Nanni O, Zoli W, Amadori D, et al. Accuracy of urine telomerase activity to detect bladder cancer in symptomatic patients. Int J Biol Markers. 2009 Oct-Dec;24(4):253-7.
17.Lai Y, Ye J, Chen J, Zhang L, Wasi L, He Z, et al. UPK3A: a promising novel urinary marker for the detection of bladder cancer. Urology. 2010 Aug;76(2):514 e6-11.
18.Kelly JD, Dudderidge TJ, Wollenschlaeger A, Okoturo O, Burling K, Tulloch F, et al. Bladder cancer diagnosis and identification of clinically significant disease by combined urinary detection of Mcm5 and nuclear matrix protein 22. PLoS One. 2012;7(7):e40305.
19.Hazzaa SM, Elashry OM, Afifi IK. Clusterin as a diagnostic and prognostic marker for transitional cell carcinoma of the bladder. Pathol Oncol Res. 2010 Mar;16(1):101-9.