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Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Genet. | doi: 10.3389/fgene.2019.01237

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 Batista1, 2, 3, 4*,  João Vinagre1, 2, 3, 4,  Hugo Prazeres1, 2, 4, 5, 6, 7,  Cristina Sampaio2, Pedro Peralta5, Paulo Conceição5, Amilcar Sismeiro5,  Ricardo Leão8, 9, Andreia Gomes8, Frederico Furriel9, Carlos Oliveira8, João N. Torres8, Pedro Pedro Eufrásio9, Paulo Azinhais9, Fábio Almeida10,  Edwin R. Gonzalez11,  Bohdan Bidovanets12,  Thorsten Ecke13,  P. Minnee14, Álvaro S. Pascual15,  Rabehi Abdelmalek16, Ainara Villafruela17,  Pastora Beardo-Villar18, Nuno Fidalgo19,  Hakan Öztürk20, Carmen Gonzalez-Enguita21, Juan Juan Monzo21, Tomé Lopes22, Luis Martínez-Piñeiro23,  Patricia P. Servan24, Santiago P. De La Cruz25,  Mario S. Perez25,  Valdemar Máximo1, 2, 3 and  Paula Soares1, 2, 3, 4, 5
  • 1Institute of Pathology and Molecular Immunology, Abel Salazar Institute of Biomedical Sciences, University of Porto, Portugal
  • 2Institute of Research and Innovation in Health, University of Porto, Portugal
  • 3Faculty of Medicine, University of Porto, Portugal
  • 4Other, Portugal
  • 5Department of Pathology, Faculty of Medicine, University of Porto, Portugal
  • 6Serviço de Urologia, Instituto Português de Oncologia, Portugal
  • 7Portuguese Oncology Institute, Portugal
  • 8Hospital Braga, Portugal
  • 9Hospital CUF Coimbra, Portugal
  • 10Hospital Universitário Fernando Pessoa, Portugal
  • 11Other, Spain
  • 12Other, Ukraine
  • 13Helios Hospital, Germany
  • 14LangeLand Hospital, Netherlands
  • 15Servicio de Urología, Hospital Clínico San Carlos, Spain
  • 16Other, Algeria
  • 17Hospital Universitario Donostia, Spain
  • 18Araba University Hospital, Spain
  • 19Hospital Garcia de Orta, Portugal
  • 20Department of Urology, Medicalpark Izmir Hospital, Turkey
  • 21University Hospital Fundación Jiménez Díaz, Spain
  • 22Hospital of Santa Maria, University Hospital Center Lisbon Norte, Portugal
  • 23University Hospital La Paz, Spain
  • 24Hospital de Mérida, Spain
  • 25University Hospital of Badajoz, Spain

Bladder cancer (BC), the most frequent malignancy of the urinary system, is ranked the sixth most prevalent cancer worldwide. 70-75% of all newly diagnosed patients with BC 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 non-invasive 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 findings, we developed an ultra-sensitive, urine-based assay called Uromonitor®, capable of detecting trace amounts of TERT promoter and FGFR3 hotspot mutations, in tumor cells exfoliated to urine samples. Cells present in urine were concentrated by the filtration of urine through filters were 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 NMBIC 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 access 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 and specific in detecting recurrence of BC in patients under surveillance of NMIBC.

Keywords: Non-muscle invasive bladder cancer, TERT promoter mutation, FGFR3 mutation, Urinary test, Uromonitor

Received: 03 Jun 2019; Accepted: 08 Nov 2019.

Copyright: © 2019 Batista, Vinagre, Prazeres, Sampaio, Peralta, Conceição, Sismeiro, Leão, Gomes, Furriel, Oliveira, Torres, Pedro Eufrásio, Azinhais, Almeida, Gonzalez, Bidovanets, Ecke, Minnee, Pascual, Abdelmalek, Villafruela, Beardo-Villar, Fidalgo, Öztürk, Gonzalez-Enguita, Juan Monzo, Lopes, Martínez-Piñeiro, Servan, De La Cruz, Perez, Máximo and Soares. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Rui Batista, Institute of Pathology and Molecular Immunology, Abel Salazar Institute of Biomedical Sciences, University of Porto, Porto, 4200-135, Portugal,