Estrogen Receptor-Beta2 (ERβ2)–Mutant p53–FOXM1 Axis: A Novel Driver of Proliferation, Chemoresistance, and Disease Progression in High Grade Serous Ovarian Cancer (HGSOC)

Simple Summary High grade serous ovarian cancer (HGSOC) is the most common and lethal subtype of ovarian cancer without effective therapeutic options. The high prevalence of mutations (~96%) in tumor suppressor p53 is a hallmark of HGSOC. Estrogen receptor-beta (ERβ) has been reported to be another important player in HGSOC, although the pro-versus anti-tumorigenic role of its different isoforms remains unclear. The aim of this study was to analyze the crosstalk between ERβ and mutant p53 and its impact on the pro-tumorigenic processes in HGSOC. Using the HGSOC cell line models and patient tumor tissue specimens, we demonstrated functional interaction between the ERβ2 isoform and mutant p53 and their ability to co-dependently increase FOXM1 gene transcription, decrease cell death, increase cell proliferation, and mediate resistance to carboplatin treatment. Furthermore, high levels of ERβ2 as well as FOXM1 correlated with worse patient survival. Collectively, our data suggest that the ERβ2-mutant p53-FOXM1 axis could be a novel therapeutic target for HGSOC. Abstract High grade serous ovarian cancer (HGSOC) is the most common and lethal subtype of epithelial ovarian cancer. Prevalence (~96%) of mutant p53 is a hallmark of HGSOC. Estrogen receptor-beta (ERβ) has been reported to be another important player in HGSOC, although the pro-versus anti-tumorigenic role of its different isoforms remains unsettled. However, whether there is functional interaction between ERβ and mutant p53 in HGSOC is unknown. ERβ1 and ERβ2 mRNA and protein analysis in HGSOC cell lines demonstrated that ERβ2 is the predominant isoform in HGSOC. Specificity of ERβ2 antibody was ascertained using cells depleted of ERβ2 and ERβ1 separately with isoform-specific siRNAs. ERβ2-mutant p53 interaction in cell lines was confirmed by co-immunoprecipitation and in situ proximity ligation assay (PLA). Expression levels of ERβ2, ERα, p53, and FOXM1 proteins and ERβ2-mutant p53 interaction in patient tumors were determined by immunohistochemistry (IHC) and PLA, respectively. ERβ2 levels correlate positively with FOXM1 levels and negatively with progression-free survival (PFS) and overall survival (OS). Quantitative chromatin immunoprecipitation (qChIP) and mRNA expression analysis revealed that ERβ2 and mutant p53 co-dependently regulated FOXM1 gene transcription. The combination of ERβ2-specific siRNA and PRIMA-1MET that converts mutant p53 to wild type conformation increased apoptosis. Our work provides the first evidence for a novel ERβ2-mutant p53-FOXM1 axis that can be exploited for new therapeutic strategies against HGSOC.

[1]  A. Saleh,et al.  Mutated p53 in HGSC—From a Common Mutation to a Target for Therapy , 2021, Cancers.

[2]  A. Karpf,et al.  FOXM1: A Multifunctional Oncoprotein and Emerging Therapeutic Target in Ovarian Cancer , 2021, Cancers.

[3]  Wei Zhang,et al.  Cellular models of development of ovarian high‐grade serous carcinoma: A review of cell of origin and mechanisms of carcinogenesis , 2021, Cell proliferation.

[4]  R. L. Hollis,et al.  Estrogen Signaling and Its Potential as a Target for Therapy in Ovarian Cancer , 2020, Cancers.

[5]  G. Mills,et al.  Clinical relevance of TP53 hotspot mutations in high-grade serous ovarian cancers , 2019, British Journal of Cancer.

[6]  D. Levine,et al.  Both fallopian tube and ovarian surface epithelium are cells-of-origin for high-grade serous ovarian carcinoma , 2019, Nature Communications.

[7]  A. Børresen-Dale,et al.  TP53 Status as a Determinant of Pro- vs Anti-Tumorigenic Effects of Estrogen Receptor-Beta in Breast Cancer , 2019, Journal of the National Cancer Institute.

[8]  R. Bast,et al.  Cell Origins of High-Grade Serous Ovarian Cancer , 2018, Cancers.

[9]  Austin Miller,et al.  Abstract B53: Estrogen receptor beta 2 (ERβ2)- p53- FOXM1 signaling axis in high-grade serous ovarian cancer (HGSOC): Underlying mechanisms and implications for resistance to therapy , 2018, Genetics and Molecular Drivers.

[10]  A. Jemal,et al.  Ovarian cancer statistics, 2018 , 2018, CA: a cancer journal for clinicians.

[11]  G. Blandino,et al.  New therapeutic strategies to treat human cancers expressing mutant p53 proteins , 2018, Journal of Experimental & Clinical Cancer Research.

[12]  S. Nilsson,et al.  Targeting estrogen receptor beta (ERβ) for treatment of ovarian cancer: importance of KDM6B and SIRT1 for ERβ expression and functionality , 2018, Oncogenesis.

[13]  R. Scharpf,et al.  High grade serous ovarian carcinomas originate in the fallopian tube , 2017, Nature Communications.

[14]  T. Curiel,et al.  Therapeutic utility of natural estrogen receptor beta agonists on ovarian cancer , 2017, Oncotarget.

[15]  O. Ortmann,et al.  Effect of estrogen receptor β agonists on proliferation and gene expression of ovarian cancer cells , 2017, BMC Cancer.

[16]  C. Crum,et al.  Serous tubal intraepithelial neoplasia: the concept and its application , 2017, Modern Pathology.

[17]  Adrian V. Lee,et al.  Active Estrogen Receptor-alpha Signaling in Ovarian Cancer Models and Clinical Specimens , 2017, Clinical Cancer Research.

[18]  R. Drapkin,et al.  Mutant p53 regulates ovarian cancer transformed phenotypes through autocrine matrix deposition. , 2016, JCI insight.

[19]  K. Wong,et al.  Mutant p53 Promotes Epithelial Ovarian Cancer by Regulating Tumor Differentiation, Metastasis, and Responsiveness to Steroid Hormones. , 2016, Cancer research.

[20]  I. Kyriakidis,et al.  Estrogen receptor beta and ovarian cancer: a key to pathogenesis and response to therapy , 2016, Archives of Gynecology and Obstetrics.

[21]  J. Gustafsson,et al.  ERβ decreases the invasiveness of triple-negative breast cancer cells by regulating mutant p53 oncogenic function , 2016, Oncotarget.

[22]  H. Hollema,et al.  Hormone receptors as a marker of poor survival in epithelial ovarian cancer. , 2015, Gynecologic oncology.

[23]  G. Scambia,et al.  Mitochondrial estrogen receptor β2 drives antiapoptotic pathways in advanced serous ovarian cancer. , 2015, Human pathology.

[24]  A. Hallberg,et al.  APR-246 overcomes resistance to cisplatin and doxorubicin in ovarian cancer cells , 2015, Cell Death and Disease.

[25]  K. Thiel,et al.  TP53 oncomorphic mutations predict resistance to platinum- and taxane-based standard chemotherapy in patients diagnosed with advanced serous ovarian carcinoma , 2014, International journal of oncology.

[26]  E. Lam,et al.  FOXM1: An emerging master regulator of DNA damage response and genotoxic agent resistance , 2014, Biochimica et biophysica acta.

[27]  Richard N. Freiman,et al.  Estrogen signaling crosstalk: Implications for endocrine resistance in ovarian cancer , 2014, The Journal of Steroid Biochemistry and Molecular Biology.

[28]  G. Scambia,et al.  Prognostic significance of the estrogen receptor beta (ERβ) isoforms ERβ1, ERβ2, and ERβ5 in advanced serous ovarian cancer. , 2014, Gynecologic oncology.

[29]  I. Sohn,et al.  Clinical Relevance of Gain-Of-Function Mutations of p53 in High-Grade Serous Ovarian Carcinoma , 2013, PloS one.

[30]  A. Whittemore,et al.  Hormone-receptor expression and ovarian cancer survival: an Ovarian Tumor Tissue Analysis consortium study. , 2013, The Lancet. Oncology.

[31]  E. Margeat,et al.  Negative regulation of estrogen signaling by ERβ and RIP140 in ovarian cancer cells. , 2013, Molecular endocrinology.

[32]  C. Sander,et al.  Evaluating cell lines as tumour models by comparison of genomic profiles , 2013, Nature Communications.

[33]  A. Sood,et al.  Estrogen receptor expression and increased risk of lymphovascular space invasion in high-grade serous ovarian carcinoma. , 2013, Gynecologic oncology.

[34]  U. Mukhopadhyay,et al.  Tumor suppressor p53 status as a determinant of estrogen receptor beta signaling in breast cancer , 2013 .

[35]  D. Tong,et al.  The prognostic value of estrogen receptor beta and proline-, glutamic acid- and leucine-rich protein 1 (PELP1) expression in ovarian cancer , 2013, BMC Cancer.

[36]  S. Schüler,et al.  Role of estrogen receptor β in gynecological cancer. , 2012, Gynecologic oncology.

[37]  P. Balaguer,et al.  Potential Role of Estrogen Receptor Beta as a Tumor Suppressor of Epithelial Ovarian Cancer , 2012, PloS one.

[38]  P. Fuller,et al.  Ovarian Actions of Estrogen Receptor-β: An Update , 2012, Seminars in Reproductive Medicine.

[39]  I. Shih,et al.  Immunohistochemical staining patterns of p53 can serve as a surrogate marker for TP53 mutations in ovarian carcinoma: an immunohistochemical and nucleotide sequencing analysis , 2011, Modern Pathology.

[40]  G. Scambia,et al.  Cytoplasmic expression of estrogen receptor beta (ERβ) predicts poor clinical outcome in advanced serous ovarian cancer. , 2011, Gynecologic oncology.

[41]  Benjamin J. Raphael,et al.  Integrated Genomic Analyses of Ovarian Carcinoma , 2011, Nature.

[42]  Lara J. Monteiro,et al.  ATM and p53 Regulate FOXM1 Expression via E2F in Breast Cancer Epirubicin Treatment and Resistance , 2011, Molecular Cancer Therapeutics.

[43]  Carlos Caldas,et al.  Driver mutations in TP53 are ubiquitous in high grade serous carcinoma of the ovary , 2010, The Journal of pathology.

[44]  A. Barsotti,et al.  Pro-proliferative FoxM1 is a target of p53-mediated repression , 2009, Oncogene.

[45]  A. Gartel,et al.  p53 negatively regulates expression of FoxM1 , 2009, Cell cycle.

[46]  J. Gustafsson,et al.  A genome-wide study of the repressive effects of estrogen receptor beta on estrogen receptor alpha signaling in breast cancer cells , 2008, Oncogene.

[47]  H. Ngan,et al.  Estrogen Receptor Subtypes in Ovarian Cancer: A Clinical Correlation , 2008, Obstetrics and gynecology.

[48]  J. Frasor,et al.  Impact of Estrogen Receptor β on Gene Networks Regulated by Estrogen Receptor α in Breast Cancer Cells , 2006 .

[49]  Galina Selivanova,et al.  Restoration of the tumor suppressor function to mutant p53 by a low-molecular-weight compound , 2002, Nature Medicine.

[50]  John O'Quigley,et al.  An application of changepoint methods in studying the effect of age on survival in breast cancer , 1999 .

[51]  Shuk-Mei Ho,et al.  Expression of human estrogen receptor-α and -β, progesterone receptor, and androgen receptor mRNA in normal and malignant ovarian epithelial cells , 1999 .

[52]  A. Børresen-Dale,et al.  TP53 Mutations in Breast and Ovarian Cancer. , 2017, Cold Spring Harbor perspectives in medicine.

[53]  J. Frasor,et al.  Impact of estrogen receptor beta on gene networks regulated by estrogen receptor alpha in breast cancer cells. , 2006, Endocrinology.

[54]  Wyeth W. Wasserman,et al.  JASPAR: an open-access database for eukaryotic transcription factor binding profiles , 2004, Nucleic Acids Res..

[55]  S. Mok,et al.  Expression of human estrogen receptor-alpha and -beta, progesterone receptor, and androgen receptor mRNA in normal and malignant ovarian epithelial cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.