Promoter hypermethylation of HS3ST2, SEPTIN9 and SLIT2 combined with FGFR3 mutations as a sensitive/specific urinary assay for diagnosis and surveillance in patients with low or high-risk non-muscle-invasive bladder cancer

BMC Cancer

Research Article

Idir Ouzaid¹  Vincent Ravery¹  François Desgrandchamps²  Pierre Mongiat-Artus²  Véronique Phe³  Calin Ciofu⁴  Olivier Cussenot⁵  Morgan Roupret⁵  Florence Tubach⁶  Jean-Pierre Roperch⁷  Bernard Grandchamp⁷  Roberto Incitti⁸ 

[1] Departement of Urology, Bichat-Claude Bernard Hospital, Paris, France;Departement of Urology, Saint-Louis Hospital, Paris, France;Department of Urology, Pitié Hospital, Paris, France;Department of Urology, Tenon Hospital, Paris, France;GRC-05, University Institute of Oncology, University Paris-6, Paris, France;Department of Urology, Pitié Hospital, Paris, France;Department of Urology, Tenon Hospital, Paris, France;INSERM, ECEVE, UMR 1123, CIC-EC 11425, Paris, France;University Paris Diderot, ECEVE, UMR 1123, Sorbonne Paris Cité, Paris, France;Department of Epidemiology and Clinical Research, Bichat-Claude Bernard Hospital, Paris, France;OncoDiag SAS, Agoranov, Paris, France;OncoDiag SAS, Agoranov, Paris, France;Computational Bioscience Research Center, KAUST University, Thuwal, Saudi Arabia;

关键词: Non-muscle-invasive bladder cancer;    Urine-based assay;    Genetic and Epigenetic DNA biomarkers;    Diagnosis;    Surveillance;   

DOI  :  10.1186/s12885-016-2748-5

 received in 2016-02-02, accepted in 2016-08-25,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundNon-muscle-invasive bladder cancer (NMIBC) is a high incidence form of bladder cancer (BCa), where genetic and epigenetic alterations occur frequently. We assessed the performance of associating a FGFR3 mutation assay and a DNA methylation analysis to improve bladder cancer detection and to predict disease recurrence of NMIBC patients.MethodsWe used allele specific PCR to determine the FGFR3 mutation status for R248C, S249C, G372C, and Y375C. We preselected 18 candidate genes reported in the literature as being hypermethylated in cancer and measured their methylation levels by quantitative multiplex-methylation specific PCR. We selected HS3ST2, SLIT2 and SEPTIN9 as the most discriminative between control and NMIBC patients and we assayed these markers on urine DNA from a diagnostic study consisting of 167 NMIBC and 105 controls and a follow-up study consisting of 158 NMIBC at diagnosis time’s and 425 at follow-up time. ROC analysis was performed to evaluate the diagnostic accuracy of each assay alone and in combination.ResultsFor Diagnosis: Using a logistic regression analysis with a model consisting of the 3 markers’ methylation values, FGFR3 status, age and known smoker status at the diagnosis time we obtained sensitivity/specificity of 97.6 %/84.8 % and an optimism-corrected AUC of 0.96. With an estimated BCa prevalence of 12.1 % in a hematuria cohort, this corresponds to a negative predictive value (NPV) of 99.6 %. For Follow-up: Using a logistic regression with FGFR3 mutation and the CMI at two time points (beginning of the follow-up and current time point), we got sensitivity/specificity/NPV of 90.3 %/65.1 %/97.0 % and a corrected AUC of 0.84. We also tested a thresholding algorithm with FGFR3 mutation and the two time points as described above, obtaining sensitivity/specificity/NPV values of, respectively, 94.5 %/75.9 %/98.5 % and an AUC of 0.82.ConclusionsWe showed that combined analysis of FGFR3 mutation and DNA methylation markers on urine can be a useful strategy in diagnosis, surveillance and for risk stratification of patients with NMIBC. These results provide the basis for a highly accurate noninvasive test for population screening and allowing to decrease the frequency of cystoscopy, an important feature for both patient quality of life improvement and care cost reduction.

【 授权许可】

CC BY   
© The Author(s). 2016