Frontiers in Genetics (Dec 2019)

Validation of a Novel, Sensitive, and Specific Urine-Based Test for Recurrence Surveillance of Patients With Non-Muscle-Invasive Bladder Cancer in a Comprehensive Multicenter Study

  • Rui Batista,
  • Rui Batista,
  • Rui Batista,
  • Rui Batista,
  • João Vinagre,
  • João Vinagre,
  • João Vinagre,
  • João Vinagre,
  • Hugo Prazeres,
  • Hugo Prazeres,
  • Hugo Prazeres,
  • Hugo Prazeres,
  • Cristina Sampaio,
  • Cristina Sampaio,
  • Pedro Peralta,
  • Paulo Conceição,
  • Amílcar Sismeiro,
  • Ricardo Leão,
  • Ricardo Leão,
  • Andreia Gomes,
  • Frederico Furriel,
  • Carlos Oliveira,
  • João Nuno Torres,
  • Pedro Eufrásio,
  • Paulo Azinhais,
  • Fábio Almeida,
  • Edwin Romero Gonzalez,
  • Bohdan Bidovanets,
  • Thorsten Ecke,
  • Pascal Stinjs,
  • Álvaro Serrano Pascual,
  • Rabehi Abdelmalek,
  • Ainara Villafruela,
  • Pastora Beardo-Villar,
  • Nuno Fidalgo,
  • Hakan Öztürk,
  • Carmen Gonzalez-Enguita,
  • Juan Monzo,
  • Tomé Lopes,
  • Mario Álvarez-Maestro,
  • Patricia Parra Servan,
  • Santiago Moreno Perez De La Cruz,
  • Mario Pual Sanchez Perez,
  • Valdemar Máximo,
  • Valdemar Máximo,
  • Valdemar Máximo,
  • Paula Soares,
  • Paula Soares,
  • Paula Soares,
  • Paula Soares

DOI
https://doi.org/10.3389/fgene.2019.01237
Journal volume & issue
Vol. 10

Abstract

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Bladder cancer (BC), the most frequent malignancy of the urinary system, is ranked the sixth most prevalent cancer worldwide. Of all newly diagnosed patients with BC, 70–75% will present disease confined to the mucosa or submucosa, the non-muscle-invasive BC (NMIBC) subtype. Of those, approximately 70% will recur after transurethral resection (TUR). Due to high rate of recurrence, patients are submitted to an intensive follow-up program maintained throughout many years, or even throughout life, resulting in an expensive follow-up, with cystoscopy being the most cost-effective procedure for NMIBC screening. Currently, the gold standard procedure for detection and follow-up of NMIBC is based on the association of cystoscopy and urine cytology. As cystoscopy is a very invasive approach, over the years, many different noninvasive assays (both based in serum and urine samples) have been developed in order to search genetic and protein alterations related to the development, progression, and recurrence of BC. TERT promoter mutations and FGFR3 hotspot mutations are the most frequent somatic alterations in BC and constitute the most reliable biomarkers for BC. Based on these, we developed an ultra-sensitive, urine-based assay called Uromonitor®, capable of detecting trace amounts of TERT promoter (c.1-124C > T and c.1-146C > T) and FGFR3 (p.R248C and p.S249C) hotspot mutations, in tumor cells exfoliated to urine samples. Cells present in urine were concentrated by the filtration of urine through filters where tumor cells are trapped and stored until analysis, presenting long-term stability. Detection of the alterations was achieved through a custom-made, robust, and highly sensitive multiplex competitive allele-specific discrimination PCR allowing clear interpretation of results. In this study, we validate a test for NMIBC recurrence detection, using for technical validation a total of 331 urine samples and 41 formalin-fixed paraffin-embedded tissues of the primary tumor and recurrence lesions from a large cluster of urology centers. In the clinical validation, we used 185 samples to assess sensitivity/specificity in the detection of NMIBC recurrence vs. cystoscopy/cytology and in a smaller cohort its potential as a primary diagnostic tool for NMIBC. Our results show this test to be highly sensitive (73.5%) and specific (93.2%) in detecting recurrence of BC in patients under surveillance of NMIBC.

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