A Prospective,Multicenter,Blinded Study Of Urine-based DNA Methylation Detection In Non-invasive Diagnosis For Urothelial Carcinoma

Backgroundurothelial carcinoma(UC)is currently one of the most common malignant tumors in the urinary system.Early UC has a good prognosis,but it is prone to recurrence and requires long-term and frequent follow-up.Advanced UC is prone to metastasis and has a dismal prognosis.Therefore,early diagnosis and postoperative follow-up are the key to UC diagnosis and treatment.Although there are many methods of detection of UC,urological endoscopy and biopsy are still the gold standard for UC diagnosis and postoperative follow-up,and their invasiveness and expensive price often reduce patient adherence.However,urine cytology and fluorescence in situ hybridization techniques are difficult to combine good sensitivity and specificity,and there is currently a lack of a non-invasive detection method that is simple to perform,accurate and economical.ObjectiveThe diagnostic potential of two DNA methylation markers was explored,and a UC noninvasive diagnostic model was established to explore the application value of the model in UC non-invasive diagnosis in a multi-center cohort.MethodsAfter analyzing the diagnostic potential of PCDHGB7 and Me3p261 in public databases and validated in tissues,the MRSE-q PCR platform was used to detect the level of methylation in 319 UC patients and 149 patients with benign urinary disease(BUD)from a single center.An early non-invasive detector(END)was established by linear fitting,and the stability of END was verified in 85 BUD samples and 86 UC samples from an external cohort.In addition,the application of END in the monitoring of recurrence after UC surgery was preliminarily explored in a follow-up cohort.Finally,the methylation levels of the two molecular markers were blindly detected in prospectively collected samples from a multi-center cohort to evaluate the diagnostic performance and application value of END in the multi-centers.ResultsPCDHGB7 and Me3p261 showed excellent diagnostic performance in the single-center cohort for tumor specific hypermethylation in UC samples.The diagnostic model after fitting the two has a sensitivity of 85.58% and a specificity of 94.00% in a single center,and has been externally verified.Compared with cystoscopy,END can help UC patients predict and detect small lesions that would lead to recurrence after surgery in advance.Finally,the model remains robust and stable in multi-centers,especially in early-stage tumors,whose sensitivity far exceeds that of urine cytology.ConclusionsPCDHGB7 and Me3p261 have high diagnostic value in UC non-invasive detection,and the END not only has good diagnostic performance in single-center cohort,but also has good application value in prospective and multi-center cohort.