Clinical evaluation of Bladder CARE, a new epigenetic test for bladder cancer detection in urine samples

Abstract

Background Bladder cancer (BC) is the 5th most common cancer in the USA. Non-muscle invasive bladder cancer represents about 70% of all cases and has generally a favorable outcome. However, recurrence rates as high as 60 to 70% and progression rates of 10 to 20% necessitate intensive surveillance with cystoscopy. The invasiveness and high cost of cystoscopy poses significant burden on BC patients as well as on the healthcare system. In this study we test the feasibility of a simple, sensitive, and non-invasive detection of BC using Bladder CARE test in urine samples. Results Urine from 136 healthy and 77 BC subjects was collected using the at-home Bladder CARE Urine Collection Kit and analyzed with Bladder CARE test. The test measures the methylation level of three BC-specific biomarkers and two internal controls using methylation-sensitive restriction enzymes coupled with qPCR. Bladder CARE showed an overall sensitivity of 93.5%, a specificity of 92.6%, and a PPV and NPV of 87.8% and 96.2%, respectively. Bladder CARE has an LOD as low as 0.046%, which equates to detecting 1 cancer cell for every 2,200 cells analyzed. We also provided evidence that bisulfite-free methods to assess DNA methylation, like Bladder CARE, are advantageous compared to conventional methods that rely on bisulfite conversion of the DNA. Conclusion Highly sensitive detection of BC in urine samples is possible using Bladder CARE. The low LOD of the test and the measurement of epigenetic biomarkers make Bladder CARE a good candidate for the early detection of BC and possibly for the routine screening and surveillance of BC patients. Bladder CARE and the at-home urine sample collection system have the potential to (1) reduce unnecessary invasive testing for BC (2) reduce the burden of surveillance on patients and on the healthcare system, (3) improve the detection of early stage BC, and (4) allow physicians to streamline the monitoring of patients.

Alterations of Chromatin Regulators in the Pathogenesis of Urinary Bladder Urothelial Carcinoma

“…DNMT3A and DNMT1 were each affected by mutations in 10 cases. This latter result suggests that DNA methylation changes in UC [ 50 , 51 , 52 , 53 , 54 ] may in some cases be caused by genetic alterations of DNA methylation writers and erasers.…”

“…Alterations of DNA methylation patterns are frequent in UC. Pathogenesis and functional consequences of these alterations and their use in bladder cancer diagnostics have been reviewed elsewhere [ 50 , 51 , 52 , 53 , 54 ].…”

Advances in Diagnosis and Therapy for Bladder Cancer

“…The Bladder CARE test uses methylation-sensitive restriction endonucleases to measure the methylation levels of three BCa-specific biomarkers, which had an overall sensitivity, specificity, PPV, and NPV of 93.5%, 92.6%, 87.8%, and 96.2%, respectively, potentially improving the detection of early BCa [77].…”

Identification of a three-miRNA panel in serum for bladder cancer diagnosis by a diagnostic test

“…It gives a huge burden to the patient family and the national healthcare system. This has greatly increased the cost of treatment and surveillance of BC ( 3 ). Also, the cystoscopy would increase the possibility of the urinary tract infection, even for flexible cystoscopy ( 4 ).…”