A Urine-Based Liquid Biopsy Method for Detection of Upper Tract Urinary Carcinoma

Abstract

BackgroundConventional clinical detection methods such as CT, urine cytology, and ureteroscopy display low sensitivity and/or are invasive in the diagnosis of upper tract urinary carcinoma (UTUC), a factor precluding their use. Previous studies on urine biopsy have not shown satisfactory sensitivity and specificity in the application of both gene mutation or gene methylation panels. Therefore, these unfavorable factors call for an urgent need for a sensitive and non-invasive method for the diagnosis of UTUC.MethodsIn this study, a total of 161 hematuria patients were enrolled with (n = 69) or without (n = 92) UTUC. High-throughput sequencing of 17 genes and methylation analysis for ONECUT2 CpG sites were combined as a liquid biopsy test panel. Further, a logistic regression prediction model that contained several significant features was used to evaluate the risk of UTUC in these patients.ResultsIn total, 86 UTUC− and 64 UTUC+ case samples were enrolled for the analysis. A logistic regression analysis of significant features including age, the mutation status of TERT promoter, and ONECUT2 methylation level resulted in an optimal model with a sensitivity of 94.0%, a specificity of 93.1%, the positive predictive value of 92.2% and a negative predictive value of 94.7%. Notably, the area under the curve (AUC) was 0.957 in the training dataset while internal validation produced an AUC of 0.962. It is worth noting that during follow-up, a patient diagnosed with ureteral inflammation at the time of diagnosis exhibiting both positive mutation and methylation test results was diagnosed with ureteral carcinoma 17 months after his enrollment.ConclusionThis work utilized the epigenetic biomarker ONECUT2 for the first time in the detection of UTUC and discovered its superior performance. To improve its sensitivity, we combined the biomarker with high-throughput sequencing of 17 genes test. It was found that the selected logistic regression model diagnosed with ureteral cancer can evaluate upper tract urinary carcinoma risk of patients with hematuria and outperform other existing panels in providing clinical recommendations for the diagnosis of UTUC. Moreover, its high negative predictive value is conducive to rule to exclude patients without UTUC.

TERT promoter mutations and methylation for telomerase activation in urothelial carcinomas: New mechanistic insights and clinical significance

“…Similar observations were also reported by other groups ( 96 , 97 ). Cystoscopy is not suitable for UTUC, but urinary detection of the TERT promoter mutation together with other biomarkers similarly showed high sensitivity and specificity for screening UTUC-caused hematuria ( 38 , 88 , 98 ). In addition, uncommon UC variants may be difficult to make diagnosis due to their atypical cytology, and the assessment of TERT promoter mutations could be very helpful ( 99 ).…”

Liquid Biopsy in Pre-Metastatic Niche: From Molecular Mechanism to Clinical Application

“…5-Hydroxymethylcytosine signatures in cfDNA are highly predictive for colorectal and gastric cancer as an ideal diagnostic biomarker for human cancers, which are superior to conventional biomarkers, and comparable to the 5hydroxymethylcytosine biomarker in tissue biopsy (110). Additionally, as a liquid biopsy marker, the urine epigenetic biomarkers have manifested satisfactory sensitivity and specificity in detecting upper tract urinary carcinoma (111). The effect of extracellular vesicles and particles (EVPs) in tumor detection and determination of cancer type has been demonstrated.…”

Urine as a Source of Liquid Biopsy for Cancer

“…However, there is great variability in its sensitivity and specificity. GSTP1 methylation is a biomarker for prostate cancer [33] and ONECUT2 (One Cut Homeobox 2) is for upper ureteral carcinoma [34]. However, due to its low sensitivity, DNA methylation is recommended only in combination with other biomarkers.…”