Circulating biomarkers represent a promising approach to identify men with apparently low-risk PCa biopsy pathology, but who harbor occult potentially aggressive tumor unsuitable for Active Surveillance. The project activities in the next 2 years will be aimed at defining the clinical relevance of novel circulating biomarkers—including miRNAs, genomic lesions in cfDNA, and ECM-related molecules—associated with disease reclassification (upgrading) in PCa patients followed in the context of the PRIAS AS protocol, based on the assumption that such molecular alterations can provide risk refinement to standard clinicopathologic features and improve patient selection.

small peptides

ECM proteins

microRNAs

released by tumor cells or host cells in response to tumor development.

publications

Group leader

Nadia Zaffaroni

Group presentation

Dr. Zaffaroni’s group has longstanding expertise in translational research in prostate cancer. In particular, it gave major contribution to the identification/functional validation of microRNAs relevant to the disease either as biomarkers or therapeutic targets/tools. This has been made possible through a strict collaboration with the institutional Prostate Cancer Program, of which Dr. Zaffaroni is co-director and responsible for pre-clinical activities, and Prof. Riccardo Valdagni director and responsible for clinical activities. Prof. Valdagni has been actively involved in Active Surveillance since 2005, considerably contributing to its development as an alternative to radical therapies in low-risk/very low-risk prostate cancer patients.

 

Role in the project, experimental design

The group led by Dr. Zaffaroni will work at defining the clinical relevance of novel circulating biomarkers —including miRNAs, genomic lesions in circulating tumor (ct)-DNA, and extracellular matrix (ECM)-related molecules— in cohorts of very low-risk, potentially indolent prostate cancer (PCa) patients followed in the context of the Prostate Cancer Research International: Active Surveillance (PRIAS) protocol, based on the assumption that such molecular alterations can provide risk refinement to standard clinico-pathological features and improve patient selection.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Clinical study

PRIAS is a prospective international protocol in which selected men with low- and very low-risk PCa are managed by a follow-up strategy. Candidates for this program are men fit for curative therapy, PSA at diagnosis less than 10 ng/mL, PSA density (PSA/prostatic volume) less than 0.20, 1 or 2 biopsy cores bearing PCa, Gleason pattern score, GPS=3+3, and digital rectal examination T1c or T2a. Follow-up is scheduled as follows: PSA every 3 months, visit every 6 months, and re-biopsy at 12, 48, and 84 months. Patients are advised to discontinue AS and to switch to radical treatment when clinical stage >T2a or GPS >3+3 (upgrading) is found at re-biopsy. Monitoring of 257 patients previously enrolled in the study will continue in the next 2 years, and an additional multicenter cohort of ≥120 patients will be recruited.

 

Translational studies

Circulating miRNAs

A miRNA signature suitable to discriminate between indolent and non-indolent (upgrading) PCa in AS patients, which has been previously defined on the training (TRS; 144 patients) and testing (TES; 113 patients) sets, will be validated in a prospectively collected series of plasma from ≥120 patients followed in our Institute and in 2 other Italian Centers in the context of the PRIAS AS protocol. According to our preliminary data, in the new cohort it is expected to find 18-24 (15-20%) non-indolent (upgrading) and 96-102 indolent tumors. Circulating miRNA expression will be measured through the same OpenArray approach used in TRS and TES sets. The predictive capability of the identified signature will be assessed in terms of AUC and the performance of the corresponding classifier will be quantified with respect to the true status according to the results of the re-biopsy assessment.

Genomic lesions in ctDNA

An ultra-deep targeted sequencing approach will be used to detect genomic lesions (including those typically found in GPS=3+3 tumors as well as others characteristic of higher grade/more advanced tumors) in circulating ctDNA obtained from all 257 patients in AS belonging to the TRS and TES sets of the miRNA study. DNA extracted from lymphocytes of the same patients will be used as germline control. Specifically, such an approach will allow the detection of copy number aberrations, point mutations, and rearrangements in genes relevant to PCa. Results will be analyzed through the use of algorithms generated by the Prof. F. Demichelis’ group, at the Centre for Integrative Biology, University of Trento, and the association between the presence of specific DNA lesions and tumor reclassification (upgrading) will be assessed. By taking advantage of the measures on the same TRS and TES sets related to both miRNA expression and ctDNA lesions, an integrated algorithm will be developed to optimize patient classification.

Circulating ECM-related proteins

Results previously obtained in the project indicate that selected ECM-related proteins i) are able to discriminate breast cancer from benign breast disease patients when assessed in plasma samples (Tagliabue group), ii) are enriched in the conditioned medium of cancer-associated fibroblasts compared to normal fibroblasts from lung cancer patients (Sozzi group) and in that of in vitro TGF-β-stimulated fibroblasts compared to normal fibroblasts from PCa patients (our group), thus suggesting them as a biomarker of a proficient tumor-stroma crosstalk. Based on these findings, the expression levels of COL10A1, COL11A1, COMP, and SPARC will be measured by immunoassay in plasma samples from all 257 patients in AS belonging to the TRS and TES sets of the miRNA study. The association between the expression levels of each ECM-related protein and patient outcome then will be assessed. The availability of such further determinations on the same patient series will allow estimation of the possible added value of the ECM-related proteins to the developed classification algorithm.

The study results are expected to identify circulating biomarkers associated with PCa reclassification during AS to be exploited for improving patient stratification and, due to the minimally-invasive nature of the test (i.e., blood test), for disease monitoring. Such biomarkers could be integrated in an updated/improved model for selection of AS patients and for early identification of those at risk of reclassification, prior to the manifestation of conventional clinical/pathological markers. A more precise identification of truly indolent PCa will allow to reduce active treatments and side effects, as well as a correct prediction of disease reclassification will limit the need of repeated biopsies and the risk of related complications.

 

Staff

Nadia Zaffaroni

Biologist, Co-Director “Prostate Cancer Program”, Group Leader

Riccardo Valdagni

Radiation Oncologist, Director “Prostate Cancer Program”

Paolo Gandellini

Pharmaceutical Biotechnologist, Scientist

Denis Cominetti

Biologist, Junior Fellow

Valentina Doldi

Biologist, Junior Fellow

Rihan El Bezawy

Medical Biotechnologist, PhD Student

Stella Tinelli

Laboratory Technician

Elisa Campi

Laboratory Technician

Laura Zanesi

Administrative Assistant

 

 

Copyright © 2017 INT

Fondazione IRCCS Istituto Nazionale dei Tumori

All rights reserved

website by Studio Luvié

Circulating biomarkers represent a promising approach to identify men with apparently low-risk PCa biopsy pathology, but who harbor occult potentially aggressive tumor unsuitable for Active Surveillance. The project activities in the next 2 years will be aimed at defining the clinical relevance of novel circulating biomarkers—including miRNAs, genomic lesions in cfDNA, and ECM-related molecules—associated with disease reclassification (upgrading) in PCa patients followed in the context of the PRIAS AS protocol, based on the assumption that such molecular alterations can provide risk refinement to standard clinicopathologic features and improve patient selection.

Copyright © 2017 INT

Fondazione IRCCS Istituto Nazionale dei Tumori

Alla rights reserved

website by Studio Luvié

Project funded with

support from Airc 5x1000

Copyright © 2017 INT

Fondazione IRCCS Istituto Nazionale dei Tumori

All rights reserved

website by Studio Luvié