Front Oncol. 2019 Sep 10;9:874. doi: 10.3389/fonc.2019.00874. eCollection 2019.
Amantini C1, Morelli MB1,2, Nabissi M2, Piva F3, Marinelli O1,2, Maggi F4, Bianchi F5, Bittoni A5, Berardi R5, Giampieri R5, Santoni G2.
The interest in liquid biopsy is growing because it could represent a non-invasive prognostic or predictive tool for clinical outcome in patients with pancreatic ductal adenocarcinoma (PDAC), an aggressive and lethal disease. In this pilot study, circulating tumor cells (CTCs), CD16 positive atypical CTCs, and CTC clusters were captured and characterized in the blood of patients with PDAC before and after palliative first line chemotherapy by ScreenCell device, immunohistochemistry, and confocal microscopy analysis. Gene profiles were performed by digital droplet PCR in isolated CTCs, five primary PDAC tissues, and three different batches of RNA from normal human pancreatic tissue. Welsh’s t-test, Kaplan-Meier survival, and Univariate Cox regression analyses have been performed. Statistical analysis revealed that the presence of high CTC number in blood is a prognostic factor for poor overall survival and progression free survival in advanced PDAC patients, before and after first line chemotherapy. Furthermore, untreated PDAC patients with CTCs, characterized by high ALCAM, POU5F1B, and SMO mRNAs expression, have shorter progression free survival and overall survival compared with patients expressing the same biomarkers at low levels. Finally, high SHH mRNA levels are negatively associated to progression free survival, whereas high vimentin mRNA levels are correlated with the most favorable prognosis. By hierarchical clustering and correlation index analysis, two cluster gene signatures were identified in CTCs: the first, with high expression of VEGFA, NOTCH1, EPCAM, IHH, is the signature of PDAC patients before chemotherapy, whereas the second, with an enrichment in the expression of CD44, ALCAM, and POU5F1B stemness and pluripotency genes, is reported after palliative chemotherapy. Overall our data support the clinic value of the identification of CTC’s specific biomarkers to improve the prognosis and the therapy in advanced PDAC patients.
atypical CTC; circulating tumor cells; digital droplet PCR; gene signature; overall survival; pancreatic cancerPMID: 31552188 PMCID: PMC6746928 DOI: 10.3389/fonc.2019.00874
1School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.2School of Pharmacy, Experimental Medicine Section, University of Camerino, Camerino, Italy.3Department of Specialistic Clinical and Odontostomatological Sciences, Polytechnic University of Marche, Ancona, Italy.4Department of Molecular Medicine, Sapienza University of Rome, Rome, Italy.5Oncology Clinic, AOU Ospedali Riuniti, Polytechnic University of Marche, Ancona, Italy.