Clinical and Scientific Background
Bladder cancer (BC) is among the most common and costliest malignancies worldwide1. Although the majority of BC cases are non-
muscle invasive (NMIBC), BC is characterized by high recurrence (~70%) and progression rates (10-20%) to muscle-invasive disease
(MIBC)2-4. As such, NMIBC patients undergo life-long surveillance through invasive cystoscopy. Based on the guidelines, different
(BCG) immunotherapy or intravesical instillation of mitomycin-C/epirubicin (chemotherapy)5, while MIBC patients undergo radical
cystectomy6. Extensive genomic characterization of BC revealed high tumor heterogeneity indicating the existence of distinct disease
As novel therapeutic interventions for BC are on the rise, including immune checkpoint inhibitors targeting Programmed cell Death
(PD)-1 receptor and its ligand PD-L1, as well as cytotoxic T-lymphocyte-associated Protein 4 (CTLA4), guiding intervention
through the stratification of BC patients according to the risk for relapse and/or also according to the predicted drug response
becomes even more critical in the selection of optimal treatment approach. Therefore, complementary biomarkers are still needed to
improve prognostic certainty and guide clinical intervention.
Urinary Analysis - Mass Spectrometry
Urine has been already recognized as an exceptional source of biomarkers, due to the high stability of the proteome and non-invasive
means of collection17. Moreover, urinary peptides carry substantial information not only for on-site but also for systemic events that
are related to BC and depict molecular changes linked to disease pathophysiology e.g. tumor invasion and inflammation.
Mass spectrometry-derived (CE-MS) urinary profiling data have been previously explored for detection of BC18 as well as
discrimination of non- from muscle-invasive form of BC19. More recently, two diagnostic panels, based on the same technology, were
published for BC detection (BC-116) and monitoring of recurrence (BC-106)20. In the latter, the urinary profiles were also indicative
of disease molecular changes during BC progression.
Mass spectrometry has been already applied for acquiring BC specific proteomics and metabolomics profiling data and several
prognostic markers based on proteomics and metabolomics studies have been reported in the literature, highlighting the value of
omics features in improving BC management. Urine and serum proteomic-based biomarkers, like SPARC26, SH3 domain binding
glutamic acid-rich protein like 3 (SH3BGRL3)27 have been recently reported as prognostic markers for BC. For instance,
according to recent metabolomics studies, i.e. in a first study investigating metabolic profiles of smokers and non-smokers with
BC outcome28, catechol-O-methyltransferase (COMT), iodotyrosine deiodinase (IYD), tubulin tyrosine ligase (TTL) were
correlated with BC survival, while in a study investigating population-based metabolic differences associated with BC29, high
expression of lysine demethylase 2A (KDM2A) and prolyl 3‐hydroxylase 2 (P3H2) and low expression of mitochondrial malic
enzyme 3 (ME3) was correlated with poor survival of African American BC patients.
New results from BioMedBC Study
In BioMedBC study, a 36-peptide model is sufficient to embrace the heterogeneity of BC patients and forecast an accurate prognosis.
Among the 36 peptides, the majority of sequenced peptides originated from multiple collagen fragments (mainly collagen alpha-1)
and were found associated with both, good and poor prognosis depending on a specific sequence. Based on the literature and our
barrier and collagenases, such as metalloproteinases (MMPs) degrade it to expose active sites and promote a pro-tumorigenic
microenvironment to facilitate tumor progression. Collagen cross-linking and thickening is then necessary during extracellular matrix
(ECM) remodeling and invasion. Elevated levels of urinary fibrinogen have already been reported in BC patients and associated with
association with the disease. Increased levels of PIGR were also found associated with a higher risk of relapse. PIGR is a member of
the immunoglobulin superfamily, involved in transcytosis of IgA and other immune complexes. Transcription factor forkhead box
protein D2 (FOXD2) was found related to poor prognosis. Additionally, nuclear FOXOs are known to mediate cell cycle arrest and
promote apoptosis34. With regards to BC, a recent analysis of long non-coding RNAs linked high FOXD2-AS1 expression to BC
progression and recurrence by acting on Act/E2F1 axis35. Among the peptides indicative of good prognosis and lower risk of BC
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BioMedBC is a Marie Sklodowska Curie Actions (MSCA) Individual Fellowship programme (H2020-MSCA-IF-2016)
funded by the European Union under the Horizon2020 Framework Programme (Grant Agreement:752755) and
coordinated by Mosaiques diagnostics