Evaluation of an Epigenetic Profile for the Detection of Bladder Cancer in Patients with Hematuria.
OBJECTIVE
Many patients enter the care cycle with gross or microscopic hematuria and undergo cystoscopy to rule out bladder cancer. Sensitivity of this invasive examination is limited, leaving many patients at risk for undetected cancer. To improve current clinical practice more sensitive and noninvasive screening methods should be applied.


METHODS
A total of 154 urine samples were collected from patients with hematuria, including 80 without and 74 with bladder cancer. DNA from cells in the urine was epigenetically profiled using 2 independent assays. Methylation specific polymerase chain reaction was performed on TWIST1. SNaPshot™ methylation analysis was done for different loci of OTX1 and ONECUT2. Additionally all samples were analyzed for mutation status of TERT (telomerase reverse transcriptase), PIK3CA, FGFR3 (fibroblast growth factor receptor 3), HRAS, KRAS and NRAS.


RESULTS
The combination of TWIST1, ONECUT2 (2 loci) and OTX1 resulted in the best overall performing panel. Logistic regression analysis on these methylation markers, mutation status of FGFR3, TERT and HRAS, and patient age resulted in an accurate model with 97% sensitivity, 83% specificity and an AUC of 0.93 (95% CI 0.88-0.98). Internal validation led to an optimism corrected AUC of 0.92. With an estimated bladder cancer prevalence of 5% to 10% in a hematuria cohort the assay resulted in a 99.6% to 99.9% negative predictive value.


CONCLUSIONS
Epigenetic profiling using TWIST1, ONECUT2 and OTX1 results in a high sensitivity and specificity. Accurate risk prediction might result in less extensive and invasive examination of patients at low risk, thereby reducing unnecessary patient burden and health care costs.
Department of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
Kim E M
KE
van Kessel
Department of Pathology, GROW-School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands; MDxHealth, Inc., Irvine, California.
Leander
L
Van Neste
Department of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
Irene
I
Lurkin
Department of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
Ellen C
EC
Zwarthoff
MDxHealth, Inc., Irvine, California; Laboratory of Bioinformatics and Computational Genomics, Ghent University, Ghent, Belgium. Electronic address: wim.vancriekinge@mdxhealth.com.
Wim
W
Van Criekinge
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D057890:Epigenomics; D005260:Female; D020869:Gene Expression Profiling; D006417:Hematuria / Q000209:etiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D015999:Multivariate Analysis; D009154:Mutation; D012680:Sensitivity and Specificity; D001749:Urinary Bladder Neoplasms / Q000150:complications / Q000175:diagnosis* / Q000235:genetics* / Q000652:urine; D055815:Young Adult
diagnosis; epigenomics; hematuria; risk; urinary bladder neoplasms