Journal Article
. 2019 Nov; 11(1):155.
doi: 10.1186/s13148-019-0750-x.

Region-specific glucocorticoid receptor promoter methylation has both positive and negative prognostic value in patients with estrogen receptor-positive breast cancer

Hilary Snider 1 Brithica Villavarajan 1 Yingwei Peng 2 Lois E Shepherd 3 Andrew C Robinson 4 Christopher R Mueller 5 
Affiliations
  • PMID: 31675993
  •     50 References
  •     3 citations

Abstract

Background: The glucocorticoid receptor (NR3C1, GR) is frequently downregulated in breast tumors, and evidence suggests it acts as a tumor suppressor in estrogen receptor-positive (ER+) breast cancer. We previously found that methylation of the GR promoter CpG island represses gene expression and occurs in ER+ breast tumors. In this study, the prognostic and predictive value of GR methylation was examined in ER+ patients from the CCTG MA.12 clinical trial of tamoxifen versus placebo in women with early breast cancer.

Methods: We developed a targeted multiplex bisulfite next-generation sequencing assay to detect methylation at multiple GR promoter regions in DNA from formalin-fixed paraffin-embedded (FFPE) samples. Following validation in a small cohort of breast tumors, ER+ FFPE tumor samples from MA.12 (n = 208) were tested. Survival analyses evaluated the impact of GR promoter methylation on patient overall survival (OS) and disease-free survival (DFS).

Results: An analysis of TCGA data found that GR methylation is prevalent in ER+ tumors and is associated with decreased gene expression and analysis of public microarray data (KM Plotter) linked decreased GR expression to a poor outcome. In MA.12, two GR promoter regions (U and C) each had prognostic value, but with opposite effects on the outcome. U methylation was associated with poor OS (HR = 1.79, P = 0.041) whereas C methylation was associated with better OS (HR = 0.40, P = 0.040) and DFS (HR = 0.49, P = 0.037). The classification of patients based on the methylation status of the two regions was prognostic for OS (P = 0.006) and DFS (P = 0.041) and revealed a group of patients (U methylated, C unmethylated) with very poor outcomes. Placebo-treated patients in this high-risk group had worse OS (HR = 2.86, P = 0.002) and DFS (HR = 2.09, P = 0.014) compared to the rest of the cohort.

Conclusion: Region-specific GR promoter methylation was an independent prognostic marker for patient survival and identified a subset of patients with poor prognosis, particularly without tamoxifen treatment. These findings provide a foundation for future studies into GR methylation as a promising prognostic biomarker in ER+ breast cancer.

Keywords: Bisulfite sequencing; Breast cancer; DNA methylation; Glucocorticoid receptor; Prognosis.

5'-heterogeneity of glucocorticoid receptor messenger RNA is tissue specific: differential regulation of variant transcripts by early-life events.
J A McCormick, V Lyons, +9 authors, K E Chapman.
Mol Endocrinol, 2000 Apr 19; 14(4). PMID: 10770488
The unliganded glucocorticoid receptor positively regulates the tumor suppressor gene BRCA1 through GABP beta.
Heather D Ritter, Lilia Antonova, Christopher R Mueller.
Mol Cancer Res, 2012 Feb 14; 10(4). PMID: 22328717
Transcriptional control of the glucocorticoid receptor: CpG islands, epigenetics and more.
Jonathan D Turner, Simone R Alt, +4 authors, Claude P Muller.
Biochem Pharmacol, 2010 Jul 06; 80(12). PMID: 20599772
Review.
Epigenetics and Precision Oncology.
Rachael J Werner, Andrew D Kelly, Jean-Pierre J Issa.
Cancer J, 2017 Sep 20; 23(5). PMID: 28926426    Free PMC article.
Review.
Biological determinants of endocrine resistance in breast cancer.
Elizabeth A Musgrove, Robert L Sutherland.
Nat Rev Cancer, 2009 Aug 25; 9(9). PMID: 19701242
Highly Cited. Review.
Molecular stratification of triple-negative breast cancers.
Charles M Perou.
Oncologist, 2010 Dec 16; 15 Suppl 5. PMID: 21138954
Glucocorticoid Receptor:MegaTrans Switching Mediates the Repression of an ERα-Regulated Transcriptional Program.
Feng Yang, Qi Ma, +10 authors, Michael G Rosenfeld.
Mol Cell, 2017 May 06; 66(3). PMID: 28475868    Free PMC article.
A 50-gene intrinsic subtype classifier for prognosis and prediction of benefit from adjuvant tamoxifen.
Stephen K Chia, Vivien H Bramwell, +12 authors, Torsten O Nielsen.
Clin Cancer Res, 2012 Jun 20; 18(16). PMID: 22711706    Free PMC article.
Highly Cited.
MethPrimer: designing primers for methylation PCRs.
Long-Cheng Li, Rajvir Dahiya.
Bioinformatics, 2002 Nov 09; 18(11). PMID: 12424112
Highly Cited.
Activation of the glucocorticoid receptor is associated with poor prognosis in estrogen receptor-negative breast cancer.
Deng Pan, Masha Kocherginsky, Suzanne D Conzen.
Cancer Res, 2011 Aug 27; 71(20). PMID: 21868756    Free PMC article.
Highly Cited.
Glucocorticoid receptor expression in breast cancer associates with older patient age.
Larissa Belova, Bertha Delgado, +3 authors, Suzanne D Conzen.
Breast Cancer Res Treat, 2008 Aug 01; 116(3). PMID: 18668364    Free PMC article.
A DNA methylation-based definition of biologically distinct breast cancer subtypes.
Olafur A Stefansson, Sebastian Moran, +9 authors, Manel Esteller.
Mol Oncol, 2014 Dec 04; 9(3). PMID: 25468711    Free PMC article.
Highly Cited.
The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2016 update.
Enis Afgan, Dannon Baker, +17 authors, Jeremy Goecks.
Nucleic Acids Res, 2016 May 04; 44(W1). PMID: 27137889    Free PMC article.
Highly Cited.
Clinical utility of gene-expression signatures in early stage breast cancer.
Maryann Kwa, Andreas Makris, Francisco J Esteva.
Nat Rev Clin Oncol, 2017 Jun 01; 14(10). PMID: 28561071
Highly Cited. Review.
Aromatase inhibitors versus tamoxifen in early breast cancer: patient-level meta-analysis of the randomised trials.
Early Breast Cancer Trialists' Collaborative Group (EBCTCG).
Lancet, 2015 Jul 28; 386(10001). PMID: 26211827
Highly Cited.
Luminal B breast cancer subtype displays a dicotomic epigenetic pattern.
Naiara G Bediaga, Elena Beristain, +6 authors, Marian M de Pancorbo.
Springerplus, 2016 Jun 23; 5. PMID: 27330889    Free PMC article.
DNA degrades during storage in formalin-fixed and paraffin-embedded tissue blocks.
Alice Guyard, Alice Boyez, +8 authors, Karen Leroy.
Virchows Arch, 2017 Aug 16; 471(4). PMID: 28812131
Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials.
Early Breast Cancer Trialists' Collaborative Group (EBCTCG).
Lancet, 2005 May 17; 365(9472). PMID: 15894097
Highly Cited.
Prognostic value of PAM50 and risk of recurrence score in patients with early-stage breast cancer with long-term follow-up.
Hege O Ohnstad, Elin Borgen, +11 authors, Bjørn Naume.
Breast Cancer Res, 2017 Nov 16; 19(1). PMID: 29137653    Free PMC article.
Biomarker assessment and molecular testing for prognostication in breast cancer.
Zuzana Kos, David J Dabbs.
Histopathology, 2016 Jan 16; 68(1). PMID: 26768030
Review.
GR and ER Coactivation Alters the Expression of Differentiation Genes and Associates with Improved ER+ Breast Cancer Outcome.
Diana C West, Deng Pan, +7 authors, Suzanne D Conzen.
Mol Cancer Res, 2016 May 04; 14(8). PMID: 27141101    Free PMC article.
BiQ Analyzer HiMod: an interactive software tool for high-throughput locus-specific analysis of 5-methylcytosine and its oxidized derivatives.
Daniel Becker, Pavlo Lutsik, +3 authors, Jörn Walter.
Nucleic Acids Res, 2014 May 31; 42(Web Server issue). PMID: 24875479    Free PMC article.
Coactivators enable glucocorticoid receptor recruitment to fine-tune estrogen receptor transcriptional responses.
Michael J Bolt, Fabio Stossi, +3 authors, Michael A Mancini.
Nucleic Acids Res, 2013 Feb 28; 41(7). PMID: 23444138    Free PMC article.
An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients.
Balazs Györffy, Andras Lanczky, +4 authors, Zoltan Szallasi.
Breast Cancer Res Treat, 2009 Dec 19; 123(3). PMID: 20020197
Highly Cited.
Transcriptional control of the human glucocorticoid receptor: identification and analysis of alternative promoter regions.
Lei Cao-Lei, Salomon Carlos Leija, +4 authors, Claude P Muller.
Hum Genet, 2011 Jan 15; 129(5). PMID: 21234764
Family Matters: Collaboration and Conflict Among the Steroid Receptors Raises a Need for Group Therapy.
Matthew J Sikora.
Endocrinology, 2016 Nov 12; 157(12). PMID: 27835038    Free PMC article.
Review.
Glucocorticoid receptor modulation decreases ER-positive breast cancer cell proliferation and suppresses wild-type and mutant ER chromatin association.
Eva Tonsing-Carter, Kyle M Hernandez, +10 authors, Suzanne D Conzen.
Breast Cancer Res, 2019 Jul 26; 21(1). PMID: 31340854    Free PMC article.
Signaling pathways and steroid receptors modulating estrogen receptor α function in breast cancer.
Rasmus Siersbæk, Sanjeev Kumar, Jason S Carroll.
Genes Dev, 2018 Sep 06; 32(17-18). PMID: 30181360    Free PMC article.
Review.
Clinical and biological significance of glucocorticoid receptor (GR) expression in breast cancer.
Rezvan Abduljabbar, Ola H Negm, +10 authors, Ian O Ellis.
Breast Cancer Res Treat, 2015 Mar 13; 150(2). PMID: 25762479
Prognostic and predictive biomarkers in breast cancer: Past, present and future.
Andrea Nicolini, Paola Ferrari, Michael J Duffy.
Semin Cancer Biol, 2017 Sep 09; 52(Pt 1). PMID: 28882552
Highly Cited. Review.
Interaction of glucocorticoid receptor (GR) with estrogen receptor (ER) α and activator protein 1 (AP1) in dexamethasone-mediated interference of ERα activity.
Sudipan Karmakar, Yetao Jin, Akhilesh K Nagaich.
J Biol Chem, 2013 Jul 03; 288(33). PMID: 23814048    Free PMC article.
Development and verification of the PAM50-based Prosigna breast cancer gene signature assay.
Brett Wallden, James Storhoff, +16 authors, Joel S Parker.
BMC Med Genomics, 2015 Aug 25; 8. PMID: 26297356    Free PMC article.
Highly Cited.
Structural underpinnings of oestrogen receptor mutations in endocrine therapy resistance.
John A Katzenellenbogen, Christopher G Mayne, +2 authors, Sarat Chandarlapaty.
Nat Rev Cancer, 2018 Apr 18; 18(6). PMID: 29662238    Free PMC article.
Highly Cited. Review.
Glucocorticoid receptor changes its cellular location with breast cancer development.
Isabel Conde, Ricardo Paniagua, +2 authors, Maria I Arenas.
Histol Histopathol, 2007 Oct 24; 23(1). PMID: 17952860
Epigenetic Biomarkers of Breast Cancer Risk: Across the Breast Cancer Prevention Continuum.
Mary Beth Terry, Jasmine A McDonald, +2 authors, Regina M Santella.
Adv Exp Med Biol, 2016 Mar 19; 882. PMID: 26987530    Free PMC article.
Review.
Deciphering Steroid Receptor Crosstalk in Hormone-Driven Cancers.
Thu H Truong, Carol A Lange.
Endocrinology, 2018 Oct 12; 159(12). PMID: 30307542    Free PMC article.
Review.
Estrogen receptor, progesterone receptor, and glucocorticoid receptor expression in normal breast tissue, breast in situ carcinoma, and invasive breast cancer.
Frederic Buxant, Corine Engohan-Aloghe, Jean-Christophe Noël.
Appl Immunohistochem Mol Morphol, 2009 Oct 31; 18(3). PMID: 19875955
An Integrated TCGA Pan-Cancer Clinical Data Resource to Drive High-Quality Survival Outcome Analytics.
Jianfang Liu, Tara Lichtenberg, +13 authors, Hai Hu.
Cell, 2018 Apr 07; 173(2). PMID: 29625055    Free PMC article.
Highly Cited.
Glucocorticoid receptor-mediated protection from apoptosis is associated with induction of the serine/threonine survival kinase gene, sgk-1.
C A Mikosz, D R Brickley, +2 authors, S D Conzen.
J Biol Chem, 2001 Mar 30; 276(20). PMID: 11278764
Differential expression of glucocorticoid receptor in human breast tissues and related neoplasms.
H-C Lien, Y-S Lu, +6 authors, Y-T Yao.
J Pathol, 2006 Apr 28; 209(3). PMID: 16639692
Clinical evidence supporting genomic tests in early breast cancer: Do all genomic tests provide the same information?
C Markopoulos, C van de Velde, +2 authors, R Masetti.
Eur J Surg Oncol, 2016 Sep 19; 43(5). PMID: 27639633
Review.
Tamoxifen for early breast cancer: an overview of the randomised trials. Early Breast Cancer Trialists' Collaborative Group.
Lancet, 1998 May 30; 351(9114). PMID: 9605801
Highly Cited.
Expression microarray identifies the unliganded glucocorticoid receptor as a regulator of gene expression in mammary epithelial cells.
Heather D Ritter, Christopher R Mueller.
BMC Cancer, 2014 Apr 24; 14. PMID: 24755251    Free PMC article.
Stress and breast cancer: from epidemiology to molecular biology.
Lilia Antonova, Kristan Aronson, Christopher R Mueller.
Breast Cancer Res, 2011 May 18; 13(2). PMID: 21575279    Free PMC article.
Review.
Reprogramming the chromatin landscape: interplay of the estrogen and glucocorticoid receptors at the genomic level.
Tina B Miranda, Ty C Voss, +6 authors, Gordon L Hager.
Cancer Res, 2013 Jun 28; 73(16). PMID: 23803465    Free PMC article.
Evaluation of bisulfite kits for DNA methylation profiling in terms of DNA fragmentation and DNA recovery using digital PCR.
Sam Kint, Ward De Spiegelaere, +2 authors, Wim Van Criekinge.
PLoS One, 2018 Jun 15; 13(6). PMID: 29902267    Free PMC article.
Frequent promoter hypermethylation and expression reduction of the glucocorticoid receptor gene in breast tumors.
Kirsten A Nesset, Ami M Perri, Christopher R Mueller.
Epigenetics, 2014 Mar 14; 9(6). PMID: 24622770    Free PMC article.
Glucocorticoids promote breast cancer metastasis.
Milan M S Obradović, Baptiste Hamelin, +8 authors, Mohamed Bentires-Alj.
Nature, 2019 Mar 15; 567(7749). PMID: 30867597
Highly Cited.
Relevance of breast cancer hormone receptors and other factors to the efficacy of adjuvant tamoxifen: patient-level meta-analysis of randomised trials.
Early Breast Cancer Trialists' Collaborative Group (EBCTCG), C Davies, +11 authors, R Peto.
Lancet, 2011 Aug 02; 378(9793). PMID: 21802721    Free PMC article.
Highly Cited.
A randomized placebo-controlled study of tamoxifen after adjuvant chemotherapy in premenopausal women with early breast cancer (National Cancer Institute of Canada--Clinical Trials Group Trial, MA.12).
V H C Bramwell, K I Pritchard, +8 authors, L Shepherd.
Ann Oncol, 2009 Jul 25; 21(2). PMID: 19628570    Free PMC article.
Glucocorticoid Receptor: A Multifaceted Actor in Breast Cancer.
Lara Malik Noureddine, Olivier Trédan, +3 authors, Coralie Poulard.
Int J Mol Sci, 2021 May 01; 22(9). PMID: 33923160    Free PMC article.
Review.
MicroRNA-19b Downregulates NR3C1 and Enhances Oxaliplatin Chemoresistance in Colon Cancer via the PI3K/AKT/mTOR Pathway.
Zhongbo Han, Chao Zhang, +4 authors, Chunxia Yang.
Clin Med Insights Oncol, 2021 May 22; 15. PMID: 34017210    Free PMC article.
A seven-nuclear receptor-based prognostic signature in breast cancer.
F Wu, W Chen, +4 authors, X Zhang.
Clin Transl Oncol, 2020 Nov 20; 23(7). PMID: 33210236