Deep Sequencing of Urinary RNAs for Bladder Cancer Molecular Diagnostics

Abstract

Purpose The majority of bladder cancer patients present with localized disease and are managed by transurethral resection. However, the high rate of recurrence necessitates lifetime cystoscopic surveillance. Developing a sensitive and specific urine-based test would significantly improve bladder cancer screening, detection, and surveillance. Experimental Design RNA-seq was used for biomarker discovery to directly assess the gene expression profile of exfoliated urothelial cells in urine derived from bladder cancer patients (n=13) and controls (n=10). Eight bladder cancer specific and 3 reference genes identified by RNA-seq were quantitated by qPCR in a training cohort of 102 urine samples. A diagnostic model based on the training cohort was constructed using multiple logistic regression. The model was further validated in an independent cohort of 101 urines. Results 418 genes were found to be differentially expressed between bladder cancer and controls. Validation of a subset of these genes was used to construct an equation for computing a probability of bladder cancer score (PBC) based on expression of 3-markers (ROBO1, WNT5A, and CDC42BPB). Setting PBC=0.45 as the cutoff for a positive test, urine testing using the 3-marker panel had overall 88% sensitivity and 92% specificity in the training cohort. The accuracy of the 3-marker panel in the independent validation cohort yielded an area under the curve of 0.87 and overall 83% sensitivity and 89% specificity. Conclusions Urine-based molecular diagnostics using this 3-marker signature could provide a valuable adjunct to cystoscopy and may lead to a reduction of unnecessary procedures for bladder cancer diagnosis.

Development of a noninvasive tool to preoperatively evaluate the muscular invasiveness of bladder cancer using a radiomics approach

“…Other noninvasive diagnostic methods, such as urine cytology examination, urine tumor DNA, urinary RNA, and circulating tumor DNA, are also promising techniques that perhaps can be combined to help solve the current dilemmas. [43][44][45] In conclusion, we propose a radiomic-clinical nomogram for the individualized preoperative differentiation of MIBC and NMIBC. This reliable, noninvasive tool can increase the accuracy of clinical T staging in patients with BCa, which may aid in clinical decision making and improve patient prognosis.…”

“…Efforts should be taken to improve the performance of our proposed nomogram. Other noninvasive diagnostic methods, such as urine cytology examination, urine tumor DNA, urinary RNA, and circulating tumor DNA, are also promising techniques that perhaps can be combined to help solve the current dilemmas …”

MicroRNA-374a Inhibits Aggressive Tumor Biological Behavior in Bladder Carcinoma by Suppressing Wnt/β-Catenin Signaling

“…Some groups found that WNT5A levels were positively correlated with the histological grade and clinical stage of bladder cancer and that the WNT5A mRNA levels varied widely among different cell lines [25]. Furthermore, aberrant expression of CDC42BPB, WNT5A, and ROBO1 could be used for the molecular diagnosis of bladder cancer [34]. Our study demonstrated that WNT5A may act as a potential biomarker and tumor-promoting factor in bladder cancer.…”

CircRNA_0058063 functions as a ceRNA in bladder cancer progression via targeting miR-486-3p/FOXP4 axis

“…Bladder cancer (BC) was ranked as the main factors that are responsible for the cancer-related deaths worldwide, with roughly 74,000 developed cases in the United States in 2015 [1,2]. The risk of BC increases with age, peaking between 50 and 70 years old, and men are three times higher than women [3].…”