Phosphorylated and Sumoylation-deficient Progesterone Receptors Drive Proliferative Gene Signatures During Breast Cancer Progression

Overview

Journal Breast Cancer Res

Specialty Oncology

Date 2012 Jun 16

PMID 22697792

Citations 61

Authors

Todd P Knutson

Andrea R Daniel

Danhua Fan

Kevin At Silverstein

Kyle R Covington

Suzanne Aw Fuqua

Carol A Lange

Affiliations

Soon will be listed here.

Abstract

Introduction: Progesterone receptors (PR) are emerging as important breast cancer drivers. Phosphorylation events common to breast cancer cells impact PR transcriptional activity, in part by direct phosphorylation. PR-B but not PR-A isoforms are phosphorylated on Ser294 by mitogen activated protein kinase (MAPK) and cyclin dependent kinase 2 (CDK2). Phospho-Ser294 PRs are resistant to ligand-dependent Lys388 SUMOylation (that is, a repressive modification). Antagonism of PR small ubiquitin-like modifier (SUMO)ylation by mitogenic protein kinases suggests a mechanism for derepression (that is, transcriptional activation) of target genes. As a broad range of PR protein expression is observed clinically, a PR gene signature would provide a valuable marker of PR contribution to early breast cancer progression.

Methods: Global gene expression patterns were measured in T47D and MCF-7 breast cancer cells expressing either wild-type (SUMOylation-capable) or K388R (SUMOylation-deficient) PRs and subjected to pathway analysis. Gene sets were validated by RT-qPCR. Recruitment of coregulators and histone methylation levels were determined by chromatin immunoprecipitation. Changes in cell proliferation and survival were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and western blotting. Finally, human breast tumor cohort datasets were probed to identify PR-associated gene signatures; metagene analysis was employed to define survival rates in patients whose tumors express a PR gene signature.

Results: 'SUMO-sensitive' PR target genes primarily include genes required for proliferative and pro-survival signaling. DeSUMOylated K388R receptors are preferentially recruited to enhancer regions of derepressed genes (that is, MSX2, RGS2, MAP1A, and PDK4) with the steroid receptor coactivator, CREB-(cAMP-response element-binding protein)-binding protein (CBP), and mixed lineage leukemia 2 (MLL2), a histone methyltransferase mediator of nucleosome remodeling. PR SUMOylation blocks these events, suggesting that SUMO modification of PR prevents interactions with mediators of early chromatin remodeling at 'closed' enhancer regions. SUMO-deficient (phospho-Ser294) PR gene signatures are significantly associated with human epidermal growth factor 2 (ERBB2)-positive luminal breast tumors and predictive of early metastasis and shortened survival. Treatment with antiprogestin or MEK inhibitor abrogated expression of SUMO-sensitive PR target-genes and inhibited proliferation in BT-474 (estrogen receptor (ER)+/PR+/ERBB2+) breast cancer cells.

Conclusions: We conclude that reversible PR SUMOylation/deSUMOylation profoundly alters target gene selection in breast cancer cells. Phosphorylation-induced PR deSUMOylation favors a permissive chromatin environment via recruitment of CBP and MLL2. Patients whose ER+/PR+ tumors are driven by hyperactive (that is, derepressed) phospho-PRs may benefit from endocrine (antiestrogen) therapies that contain an antiprogestin.

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References

Lenferink A, Busse D, Flanagan W, Yakes F, Arteaga C . ErbB2/neu kinase modulates cellular p27(Kip1) and cyclin D1 through multiple signaling pathways. Cancer Res. 2001; 61(17):6583-91. View

Birney E, Stamatoyannopoulos J, Dutta A, Guigo R, Gingeras T, Margulies E . Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project. Nature. 2007; 447(7146):799-816. PMC: 2212820. DOI: 10.1038/nature05874. View

Lupien M, Eeckhoute J, Meyer C, Wang Q, Zhang Y, Li W . FoxA1 translates epigenetic signatures into enhancer-driven lineage-specific transcription. Cell. 2008; 132(6):958-70. PMC: 2323438. DOI: 10.1016/j.cell.2008.01.018. View

Lange C . Challenges to defining a role for progesterone in breast cancer. Steroids. 2008; 73(9-10):914-21. PMC: 2481303. DOI: 10.1016/j.steroids.2007.12.023. View

Chlebowski R, Kuller L, Prentice R, Stefanick M, Manson J, Gass M . Breast cancer after use of estrogen plus progestin in postmenopausal women. N Engl J Med. 2009; 360(6):573-87. PMC: 3963492. DOI: 10.1056/NEJMoa0807684. View

Halford S, Marko J . How do site-specific DNA-binding proteins find their targets?. Nucleic Acids Res. 2004; 32(10):3040-52. PMC: 434431. DOI: 10.1093/nar/gkh624. View

Li X, Wong J, Tsai S, Tsai M, OMalley B . Progesterone and glucocorticoid receptors recruit distinct coactivator complexes and promote distinct patterns of local chromatin modification. Mol Cell Biol. 2003; 23(11):3763-73. PMC: 155204. DOI: 10.1128/MCB.23.11.3763-3773.2003. View

Ong C, Corces V . Enhancer function: new insights into the regulation of tissue-specific gene expression. Nat Rev Genet. 2011; 12(4):283-93. PMC: 3175006. DOI: 10.1038/nrg2957. View

Suzuki T, Miki Y, Nakamura Y, Moriya T, Ito K, Ohuchi N . Sex steroid-producing enzymes in human breast cancer. Endocr Relat Cancer. 2005; 12(4):701-20. DOI: 10.1677/erc.1.00834. View

10.

Paik S, Shak S, Tang G, Kim C, Baker J, Cronin M . A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer. N Engl J Med. 2004; 351(27):2817-26. DOI: 10.1056/NEJMoa041588. View

11.

Geiss-Friedlander R, Melchior F . Concepts in sumoylation: a decade on. Nat Rev Mol Cell Biol. 2007; 8(12):947-56. DOI: 10.1038/nrm2293. View

12.

Horwitz K, Mockus M, Lessey B . Variant T47D human breast cancer cells with high progesterone-receptor levels despite estrogen and antiestrogen resistance. Cell. 1982; 28(3):633-42. DOI: 10.1016/0092-8674(82)90218-5. View

13.

Verzi M, Shin H, He H, Sulahian R, Meyer C, Montgomery R . Differentiation-specific histone modifications reveal dynamic chromatin interactions and partners for the intestinal transcription factor CDX2. Dev Cell. 2010; 19(5):713-26. PMC: 3001591. DOI: 10.1016/j.devcel.2010.10.006. View

14.

Amit I, Citri A, Shay T, Lu Y, Katz M, Zhang F . A module of negative feedback regulators defines growth factor signaling. Nat Genet. 2007; 39(4):503-12. DOI: 10.1038/ng1987. View

15.

Su B, Wong C, Hong Y, Chen S . Growth factor signaling enhances aromatase activity of breast cancer cells via post-transcriptional mechanisms. J Steroid Biochem Mol Biol. 2010; 123(3-5):101-8. PMC: 3030665. DOI: 10.1016/j.jsbmb.2010.11.012. View

16.

Musgrove E, Sutherland R . Biological determinants of endocrine resistance in breast cancer. Nat Rev Cancer. 2009; 9(9):631-43. DOI: 10.1038/nrc2713. View

17.

J van t Veer L, Dai H, van de Vijver M, He Y, Hart A, Mao M . Gene expression profiling predicts clinical outcome of breast cancer. Nature. 2002; 415(6871):530-6. DOI: 10.1038/415530a. View

18.

Meijsing S, Pufall M, So A, Bates D, Chen L, Yamamoto K . DNA binding site sequence directs glucocorticoid receptor structure and activity. Science. 2009; 324(5925):407-10. PMC: 2777810. DOI: 10.1126/science.1164265. View

19.

Beral V . Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet. 2003; 362(9382):419-27. DOI: 10.1016/s0140-6736(03)14065-2. View

20.

Graham J, Mote P, Salagame U, van Dijk J, Balleine R, Huschtscha L . DNA replication licensing and progenitor numbers are increased by progesterone in normal human breast. Endocrinology. 2009; 150(7):3318-26. PMC: 2703536. DOI: 10.1210/en.2008-1630. View