Modulation of pancreatic cancer cell sensitivity to FOLFIRINOX through microRNA-mediated regulation of DNA damage

Pietro Carotenuto,Sergio Xavier Azevedo,Rosie Upstill-Goddard,David K Chang,Vladimir Kirkin,Ian Said‐Huntingford ,Paul Workman,Vincenza Guzzardo,Rao S ,Francesco Amato,Andrew V Biankin,Andrea Lanese,Maya Raj,Kate Young,Claudia Parisi,David Cunningham,Domenico Zito,Matteo Fassan,Aldo Scarpa,Caterina Vicentini,Colin Rae,Somaieh Hedayat,Anguraj Sadanandam,Ruwaida Begum,Kyriakos Kouvelakis,Vasiliki Michalarea,Naureen Starling,Jens C Hahne,David Watkins,Maria C Previdi,Andrew Wotherspoon,Andrea Lampis,Nigel B Jamieson,Nicola Valeri,Francesco Sclafani,Albert Hallsworth,Ian Chau,Chiara Braconi

doi:10.1038/s41467-021-27099-6

Abstract

FOLFIRINOX, a combination of chemotherapy drugs (Fluorouracil, Oxaliplatin, Irinotecan -FOI), provides the best clinical benefit in pancreatic ductal adenocarcinoma (PDAC) patients. In this study we explore the role of miRNAs (MIR) as modulators of chemosensitivity to identify potential biomarkers of response. We find that 41 and 84 microRNA inhibitors enhance the sensitivity of Capan1 and MiaPaCa2 PDAC cells respectively. These include a MIR1307-inhibitor that we validate in further PDAC cell lines. Chemotherapy-induced apoptosis and DNA damage accumulation are higher in MIR1307 knock-out (MIR1307KO) versus control PDAC cells, while re-expression of MIR1307 in MIR1307KO cells rescues these effects. We identify binding of MIR1307 to CLIC5 mRNA through covalent ligation of endogenous Argonaute-bound RNAs cross-linking immunoprecipitation assay. We validate these findings in an in vivo model with MIR1307 disruption. In a pilot cohort of PDAC patients undergoing FOLFIRONX chemotherapy, circulating MIR1307 correlates with clinical outcome.

Highlights

FOLFIRINOX, a combination of chemotherapy drugs (Fluorouracil, Oxaliplatin, Irinotecan -FOI), provides the best clinical benefit in pancreatic ductal adenocarcinoma (PDAC) patients
By using a high-throughput genome-wide approach exploring the functional role of MIRs in pancreatic cancer cells exposed to chemotherapy, we identify the role of MIR1307 in mediating chemoresistance to FOLFIRINOX and propose its use as a biomarker of response to this chemotherapy regimen
We performed a functional genome-wide highthroughput screening (HTS) of ~1000 Locked Nucleic Acid (LNA) MIR inhibitors in human Pancreatic ductal adenocarcinoma (PDAC) cells treated with a combination of fluorouracil (F), oxaliplatin (O), and irinotecan (I) that resembles the FOLFIRINOX regimen in vitro

Summary

Introduction

FOLFIRINOX, a combination of chemotherapy drugs (Fluorouracil, Oxaliplatin, Irinotecan -FOI), provides the best clinical benefit in pancreatic ductal adenocarcinoma (PDAC) patients. We find that 41 and 84 microRNA inhibitors enhance the sensitivity of Capan[1] and MiaPaCa2 PDAC cells respectively. We identify binding of MIR1307 to CLIC5 mRNA through covalent ligation of endogenous Argonaute-bound RNAs cross-linking immunoprecipitation assay. We validate these findings in an in vivo model with MIR1307 disruption. By using a high-throughput genome-wide approach exploring the functional role of MIRs in pancreatic cancer cells exposed to chemotherapy, we identify the role of MIR1307 in mediating chemoresistance to FOLFIRINOX and propose its use as a biomarker of response to this chemotherapy regimen

Methods

Results

Conclusion