External Validation of a Multiplex Urinary Protein Panel for the Detection of Bladder Cancer in a Multicenter Cohort

Abstract

Background Due to the faltering sensitivity and/or specificity, urine-based assays currently have a limited role in the management of patients with bladder cancer (BCa). The aim of this study was to externally validate our previously reported protein biomarker panel from multiple sites in the US and Europe. Methods This multicenter external validation study included a total of 320 subjects (BCa = 183). The 10 biomarkers (IL8, MMP9, MMP10, SERPINA1, VEGFA, ANG, CA9, APOE, SDC1 and SERPINE1) were measured using commercial ELISA assays in an external laboratory. The diagnostic performance of the biomarker panel was assessed using receiver operator curves (ROC) and descriptive statistical values. Results Utilizing the combination of all 10 biomarkers, the AUROC for the diagnostic panel was noted to be 0.847 [95% CI: 0.796 – 0.899], outperforming any single biomarker. The multiplex assay at optimal cutoff value achieved an overall sensitivity of 0.79, specificity of 0.79, PPV of 0.73 and NPV of 0.84 for BCa classification. Sensitivity values of the diagnostic panel for high-grade BCa, low-grade BCa, MIBC and NMIBC were 0.81, 0.90, 0.95 and 0.77, respectively. Conclusions Urinary levels of the biomarker panel enabled discrimination of BCa patients and controls, and the levels of biomarker subsets were associated with advancing tumor grade and stage. Impact If proven to be reliable, urinary diagnostic biomarker assays can detect BCa in a timely manner such that the patient can expect improvements in overall survival and quality of life.

Development and Validation of Urine-based Peptide Biomarker Panels for Detecting Bladder Cancer in a Multi-center Study

“…A direct comparison of different markers and their performance, as reported in various studies is difficult, mainly due to differences in the clinical design of the respective studies. A 10-biomarker ELISA-based assay (IL8, MMP9, MMP10, SERPINA1, VEGFA, ANG, CA9, APOE, SDC1, and SERPINE1) provided an overall AUC of 0.85 (95% CI, 0.80-0.91) in discriminating urothelial bladder cancer patients from healthy and benign controls (45), slightly lower than the rates received from the CE-MS classifier for primary urothelial bladder cancer (AUC ¼ 0.87; 95% CI, 0.83-0.91). A three-gene methylation panel (OTX1, ONECUT2, and OSR1) detected low/intermediate risk urothelial bladder cancer with a sensitivity of 74% and specificity of 90% (43).…”

Developing proteomic biomarkers for bladder cancer: towards clinical application

“…The potential of such an approach is demonstrated in numerous studies published by Rosser and collaborators. [69][70][71][72][73][74][75][76][77][78] A 10-biomarker panel consisting of i nterleukin-8, matrix metalloproteinase (MMP)9, MMP10, α-1-antitrypsin, vascular endothelial growth factor A, angiogenin, carbonic anhydrase 9, apolipoprotein E, syndecan-1 and plasminogen activator inhibitor 1 was developed based on the consolidation of data derived from various 'omics' approaches including transcriptomics 72,75 and proteomics 55 followed by individual validation in immunebased assays, which measured single proteins. 70,73,74,[76][77][78] The diagnostic capabilities of this panel were assessed in an external multicentre study of 320 individuals, of which 183 had primary urothelial carcinomas, 96 had benign urological disorders, including voiding symptoms, urolithiasis, UTIs and/or haematuria, and 41 healthy individuals.…”

A multiplex immunoassay for the non-invasive detection of bladder cancer.

“…We have reported the derivation and the validation of the urinary 10-biomarker BCa diagnostic signature in a number of studies with a total of over 800 subjects [13][14][15][16][17][18]. Recent work from our laboratory using quantitative immunohistochemical staining techniques for the detection of the signature proteins has shown a strong association of expression and malignancy in human bladder tumor tissues [27].…”

“…Next, we reported the validation of the 10-biomarker diagnostic panel (IL8, MMP9, MMP10, SERPINA1, VEGFA, ANG, CA9, APOE, SDC1 and SERPINE1) in a large cohort of patients (n = 308; 102 BCa and 206 controls) including controls with diverse urologic conditions (e.g., urolithiasis, moderate-severe voiding symptoms, urinary tract infection and hematuria) [17]. Recently, an outside laboratory externally validated the 10-biomarker diagnostic panel in a large cohort of patients (n = 320; 183 BCa and 137 controls) [18]. In this study, we investigated the feasibility of developing a multiplex assay that could accurately and simultaneously monitor the diagnostic biomarkers in an efficient format for potential clinical application.…”