The NRF2 transcriptional target NQO1 has low mRNA levels in TP53-mutated endometrial carcinomas.

Guillaume Beinse,Carole Nicco,Eric Pasmant,François Goldwasser,Nathaniel Edward Bennett Saidu,Bastien Rance,Franck Letourneur,Karen Leroy,Frederic Batteux,Jérôme Alexandre,Pierre-Alexandre Just,Sandrine Chouzenoux,Brigitte Izac,Bruno Borghese,Hiromu Suzuki

doi:10.1371/journal.pone.0214416

Abstract

BackgroundNRF2 is a major transcription factor regulating the expression of antioxidative/detoxifying enzymes, involved in oncogenic processes and drug resistance. We aimed to identify molecular alterations associated with NRF2 activation in endometrial carcinoma (EC).MethodsNinety patients treated (2012–2017) for localized/locally advanced EC were included in this study. Formalin-fixed paraffin-embedded tissue samples were processed for immunohistochemical (NRF2 and Mismatch Repair proteins) analyses. Next generation sequencing (NGS) of a panel of genes including POLE, TP53, NFE2L2, KEAP1 and CUL3 was performed using Ampliseq panels on Ion Torrent PGM (ThermoFisher). NRF2 activity was assessed by NQO1, GCLC, and AKR1C3 mRNA expressions, using TaqMan assays and quantitative RT-PCR.ResultsTumors were classified as POLE exonuclease domain mutated (N = 3, 3%), MMR-deficient (MSI-like) (N = 28, 31%), TP53 mutated (Copy-number high-like) (N = 22, 24%), and other tumors (Copy-number low-like) (N = 32, 36%). NRF2 nuclear immunostaining did not correlate with NRF2 target genes expression. The 3 tumors with highest NRF2 target genes expression harbored oncogenic KEAP1 or NFE2L2 mutations. Low NQO1 mRNA and protein levels were observed in the TP53 mutated subgroup compared to others tumors (p < .05) and in silico analyses of The Cancer Genome Atlas data further indicated that NQO1 mRNA levels were lower in serous compared to endometrioid copy-number high EC.ConclusionIn contrast with previous reports based on immunohistochemistry, our study indicates that NRF2 activation is a rare event in EC, associated with NFE2L2 or KEAP1 mutations. The subset of aggressive EC with low NQO1 mRNA level might represent a specific subgroup, which could be sensitive to combination therapies targeting oxidative stress.

Highlights

Endometrial carcinoma is the most frequent gynecological cancer in woman
In contrast with previous reports based on immunohistochemistry, our study indicates that NRF2 activation is a rare event in endometrial carcinoma (EC), associated with NFE2L2 or Kelch-like ECH-associated protein 1 (KEAP1) mutations
The subset of aggressive EC with low NQO1 mRNA level might represent a specific subgroup, which could be sensitive to combination therapies targeting oxidative stress

Summary

Introduction

Endometrial carcinoma is the most frequent gynecological cancer in woman. Two main histological types have been described, type 1 endometrioid carcinoma and type 2 including nonendometrioid subtypes (high grade serous, clear cell carcinoma, carcinosarcoma) with poorer prognosis [1]. This classification was refined in 2013 by an integrated genomic characterization [2], which allowed to identify four major molecular groups: 1/ an ultra-mutated group, with DNA-Polymerase ε (POLE) catalytic subunit A mutations; 2/ a hypermutated group, characterized by a somatic microsatellite instability (MSI), largely due to methylations in MLH1 promoter; 3/ a group characterized by low copy-number alterations (Copy-number low group (CNL)); and 4/ a group characterized by high copy number alterations (Copy-number high group (CNH)) and TP53 mutations, that includes most serous carcinoma and some high grade endometrioid histologic subtypes, and that carries the worst prognosis Despite these new insights, therapeutic breakthroughs are still awaited. We aimed to identify molecular alterations associated with NRF2 activation in endometrial carcinoma (EC)

Objectives

Methods

Results

Conclusion