Correlation between High PD-L1 and EMT/Invasive Genes Expression and Reduced Recurrence-Free Survival in Blood-Circulating Tumor Cells from Patients with Non-Muscle-Invasive Bladder Cancer.

Maria Beatrice Morelli,Massimo Nabissi,Federica Maggi,Giorgio Santoni,Oliviero Marinelli,Antonella Giannantoni,Rodolfo Montironi,Jacopo Adolfo Rossi De Vermandois,Consuelo Amantini,Matteo Santoni,Marilena Gubbiotti,Ettore Mearini,Alessia Cimadamore

doi:10.3390/cancers13235989

Abstract

Simple SummaryThe correlation between immune checkpoint-programmed death-ligand 1 (PD-L1) marker and epithelial–mesenchymal transition (EMT) status may help to identify potential biomarkers for the use of immune checkpoint blockades and other immunotherapy approaches in non-muscle invasive bladder cancer (NMIBC) recurrent patients. The aim of our study was to assess to potential use of PD-L1, TWIST1, ZEB1, VIMENTIN and TIMP2 mRNA expression as prognostic biomarkers.Background: PD-L1 represents a crucial immune checkpoint molecule in the tumor microenvironment, identified as a key target for cancer immunotherapy. A correlation between PD-L1 and EMT-related genes expression in various human cancers has been suggested. Methods: By ScreenCell filtration, digital droplet PCR and confocal microscopy analysis, we aimed to investigate the expression of PD-L1 and EMT/invasive genes (TWIST1, ZEB1, VIMENTIN, TIMP2) in circulating tumor cells (CTCs) collected from the blood of non-muscle-invasive bladder cancer (NMIBC) patients, assessing the prognostic value of these biomarkers in the disease. Welchs’ test and Mann–Whitney U test, correlation index, Kaplan–Meier, Univariate and Multivariate Cox hazard proportional analysis were used. Results: Higher PD-L1, TIMP2 and VIM mRNA levels were found in pT1 compared to pTa NMIBC. As evaluated by Kaplan–Meier and Univariate and Multivariate Cox analysis, enhancement of PD-L1, TWIST1 and TIMP2 expression reduces the recurrent free survival in NMIBC patients. Conclusions: High PD-L1, TWIST1 and TIMP2 mRNAs mark the recurrent-NMIBC patients and by reducing the RFS represent negative prognostic biomarkers in these patients.

Highlights

Over 400,000 patients are newly diagnosed with bladder cancer (BC) every year [1]
We found that among 48 non-muscle-invasive bladder cancer (NMIBC) patients collected by ScreenCell filtration, 5 samples were circulating tumor cells (CTCs) negative, whereas 43/48 (89.6%) had detectable CTCs, with a median CTC count of about 4.0
Based on the epithelial mesenchymal transition (EMT) markers, we demonstrated that 17.5% of NMIBC patients displayed epithelial CTCs (E-CTCs), 55% hybrid (H-CTCs) EPCAMHigh/VIMHigh or EPCAMLow/ VIMLow and 27.5% displayed mesenchymal CTCs (M-CTCs) (Table 5)

Summary

Introduction

Three quarters of them have tumors that do not invade the detrusor muscle, which are described as non-muscle-invasive bladder cancers (NMIBCs) [2]. Treatment of NMIBC patients is based on the risk of tumor recurrence and/or progression [2]. Methods: By ScreenCell filtration, digital droplet PCR and confocal microscopy analysis, we aimed to investigate the expression of PD-L1 and EMT/invasive genes (TWIST1, ZEB1, VIMENTIN, TIMP2) in circulating tumor cells (CTCs) collected from the blood of non-muscle-invasive bladder cancer (NMIBC) patients, assessing the prognostic value of these biomarkers in the disease. As evaluated by Kaplan–Meier and Univariate and Multivariate Cox analysis, enhancement of PD-L1, TWIST1 and TIMP2 expression reduces the recurrent free survival in NMIBC patients. Conclusions: High PD-L1, TWIST1 and TIMP2 mRNAs mark the recurrent-NMIBC patients and by reducing the RFS represent negative prognostic biomarkers in these patients

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