Top

Published in:

01-12-2021 | Breast Cancer | Research article

Analysis of genomic and non-genomic signaling of estrogen receptor in PDX models of breast cancer treated with a combination of the PI3K inhibitor alpelisib (BYL719) and fulvestrant

Authors: Julien Jacquemetton, Loay Kassem, Coralie Poulard, Ahmed Dahmani, Ludmilla De Plater, Elodie Montaudon, Laura Sourd, Ludivine Morisset, Rania El Botty, Sophie Chateau-Joubert, Sophie Vacher, Ivan Bièche, Isabelle Treilleux, Olivier Trédan, Elisabetta Marangoni, Muriel Le Romancer

Published in: Breast Cancer Research | Issue 1/2021

Abstract

Background

Endocrine therapies targeting estrogen signaling have significantly improved breast cancer (BC) patient survival, although 40% of ERα-positive BCs do not respond to those therapies. Aside from genomic signaling, estrogen triggers non-genomic pathways by forming a complex containing methylERα/Src/PI3K, a hallmark of aggressiveness and resistance to tamoxifen. We aimed to confirm the prognostic value of this complex and investigated whether its targeting could improve tumor response in vivo.

Methods

The interaction of ERα/Src and ERα/PI3K was studied by proximity ligation assay (PLA) in a cohort of 440 BC patients. We then treated patient-derived BC xenografts (PDXs) with fulvestrant or the PI3K inhibitor alpelisib (BYL719) alone or in combination. We analyzed their anti-proliferative effects on 6 ERα+ and 3 ERα− PDX models. Genomic and non-genomic estrogen signaling were assessed by measuring ERα/PI3K interaction by PLA and the expression of estrogen target genes by RT-QPCR, respectively.

Results

We confirmed that ERα/Src and ERα/PI3K interactions were associated with a trend to poorer survival, the latter displaying the most significant effects. In ERα+ tumors, the combination of BYL719 and fulvestrant was more effective than fulvestrant alone in 3 models, irrespective of PI3K, PTEN status, or ERα/PI3K targeting. Remarkably, resistance to fulvestrant was associated with non-genomic ERα signaling, since genomic degradation of ERα was unaltered in these tumors, whereas the treatment did not diminish the level of ERα/PI3K interaction. Interestingly, in 2 ERα− models, fulvestrant alone impacted tumor growth, and this was associated with a decrease in ERα/PI3K interaction.

Conclusions

Our results demonstrate that ERα/PI3K may constitute a new prognostic marker, as well as a new target in BC. Indeed, resistance to fulvestrant in ERα+ tumors was associated with a lack of impairment of ERα/PI3K interaction in the cytoplasm. In addition, an efficient targeting of ERα/PI3K in ERα− tumors could constitute a promising therapeutic option.

Available only for authorised users

Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;136:E359–86. https://doi.org/10.1002/ijc.29210.CrossRef

Santen RJ, Boyd NF, Chlebowski RT, Cummings S, Cuzick J, Dowsett M, et al. Critical assessment of new risk factors for breast cancer: considerations for development of an improved risk prediction model. Endocr Relat Cancer. 2007;14:169–87. https://doi.org/10.1677/ERC-06-0045.CrossRefPubMed

Musgrove EA, Sutherland RL. Biological determinants of endocrine resistance in breast cancer. Nat Rev Cancer. 2009;9:631–43. https://doi.org/10.1038/nrc2713.CrossRefPubMed

Ring A, Dowsett M. Mechanisms of tamoxifen resistance. Endocr Relat Cancer. 2004;11:643–58. https://doi.org/10.1677/erc.1.00776.CrossRefPubMed

Zhang XH, Giuliano M, Trivedi MV, Schiff R, Osborne CK. Metastasis dormancy in estrogen receptor-positive breast cancer. Clin Cancer Res. 2013;19:6389–97. https://doi.org/10.1158/1078-0432.CCR-13-0838.CrossRefPubMed

Isakoff SJ, Engelman JA, Irie HY, Luo J, Brachmann SM, Pearline RV, et al. Breast cancer-associated PIK3CA mutations are oncogenic in mammary epithelial cells. Cancer Res. 2005;65:10992–1000. https://doi.org/10.1158/0008-5472.CAN-05-2612.CrossRefPubMed

Samuels Y, Velculescu VE. Oncogenic mutations of PIK3CA in human cancers. Cell Cycle. 2004;3:1221–4. https://doi.org/10.4161/cc.3.10.1164.CrossRef

Miller TW, Hennessy BT, Gonzalez-Angulo AM, Fox EM, Mills GB, Chen H, Higham C, Garcia-Echeverria C, Shyr Y, Arteaga CL (2010) Hyperactivation of phosphatidylinositol-3 kinase promotes escape from hormone dependence in estrogen receptor-positive human breast cancer. J Clin Invest 120: 2406-2413.https://doi.org/10.1172/JCI41680

Miller TW, Balko JM, Fox EM, Ghazoui Z, Dunbier A, Anderson H, et al. ERα-dependent E2F transcription can mediate resistance to estrogen deprivation in human breast cancer. Cancer Discov. 2011;1:338–51. https://doi.org/10.1158/2159-8290.CD-11-0101.CrossRefPubMedPubMedCentral

10.

Andre F, Ciruelos E, Rubovszky G, Campone M, Loibl S, Rugo HS, et al. Alpelisib for PIK3CA-mutated, hormone receptor-positive advanced breast cancer. N Engl J Med. 2019;380:1929–40. https://doi.org/10.1056/NEJMoa1813904.CrossRefPubMed

11.

Sarbassov DD, Guertin DA, Ali SM, Sabatini DM. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science. 2005;307:1098–101. https://doi.org/10.1126/science.1106148.CrossRefPubMed

12.

Bosch A, Li Z, Bergamaschi A, Ellis H, Toska E, Prat A, et al. PI3K inhibition results in enhanced estrogen receptor function and dependence in hormone receptor-positive breast cancer. Sci Transl Med. 2015;7:283ra51. https://doi.org/10.1126/scitranslmed.aaa4442.CrossRefPubMedPubMedCentral

13.

Castoria G, Migliaccio A, Bilancio A, Di DM, De FA, Lombardi M, Fiorentino R, Varricchio L, Barone MV, Auricchio F (2001) PI3-kinase in concert with Src promotes the S-phase entry of oestradiol-stimulated MCF-7 cells. EMBO J 20: 6050-66059. https://doi.org/10.1093/emboj/20.21.6050

14.

Simoncini T, Hafezi-Moghadam A, Brazil DP, Ley K, Chin WW, Liao JK. Interaction of estrogen receptor with the regulatory subunit of phosphatidylinositol-3-OH kinase. Nature. 2000;407:538–41. https://doi.org/10.1038/35035131.CrossRefPubMedPubMedCentral

15.

Cabodi S, Moro L, Baj G, Smeriglio M, Di SP, Gippone S, et al. p130Cas interacts with estrogen receptor alpha and modulates non-genomic estrogen signaling in breast cancer cells. J Cell Sci. 2004;117:1603–11. https://doi.org/10.1242/jcs.0102.CrossRefPubMed

16.

Le Romancer M, Poulard C, Cohen P, Sentis S, Renoir JM, Corbo L. Cracking the estrogen receptor’s posttranslational code in breast tumors. Endocr Rev. 2011;32:597–622. https://doi.org/10.1210/er.2010-0016.CrossRefPubMed

17.

Song RX, Zhang Z, Santen RJ. Estrogen rapid action via protein complex formation involving ERα and Src. Trends Endocrinol Metab. 2005;16:347–53. https://doi.org/10.1016/j.tem.2005.06.010.CrossRefPubMed

18.

Le Romancer M, Treilleux I, Leconte N, Robin-Lespinasse Y, Sentis S, Bouchekioua-Bouzaghou K, et al. Regulation of estrogen rapid signaling through arginine methylation by PRMT1. Mol Cell. 2008;31:212–21. https://doi.org/10.1016/j.molcel.2008.05.025.CrossRefPubMed

19.

Le Romancer M, Treilleux I, Bouchekioua-Bouzaghou K, Sentis S, Corbo L. Methylation, a key step for nongenomic estrogen signaling in breast tumors. Steroids. 2010;75:560–4. https://doi.org/10.1016/j.steroids.2010.01.013.CrossRefPubMed

20.

Soderberg O, Gullberg M, Jarvius M, Ridderstrale K, Leuchowius KJ, Jarvius J, et al. Direct observation of individual endogenous protein complexes in situ by proximity ligation. Nat Methods. 2006;3:995–1000. https://doi.org/10.1038/nmeth947.CrossRefPubMed

21.

Poulard C, Treilleux I, Lavergne E, Bouchekioua-Bouzaghou K, Goddard-Leon S, Chabaud S, et al. Activation of rapid estrogen signaling in aggressive human breast cancers. EMBO Mol Med. 2012;4:1200–13. https://doi.org/10.1002/emmm.201201615.CrossRefPubMedPubMedCentral

22.

Poulard C, Jacquemetton J, Tredan O, Cohen PA, Vendrell J, Ghayad SE, et al. Estrogen non-genomic signaling is activated in tamoxifen-resistant breast cancer. Int J Mol Sci. 2019;20. https://doi.org/10.3390/ijms20112773.

23.

Cottu P, Marangoni E, Assayag F, de CP, Vincent-Salomon A, Guyader C, de PL, Elbaz C, Karboul N, Fontaine JJ, Chateau-Joubert S, Boudou-Rouquette P, Alran S, Dangles-Marie V, Gentien D, Poupon MF, Decaudin D (2012) Modeling of response to endocrine therapy in a panel of human luminal breast cancer xenografts. Breast Cancer Res Treat 133: 595-606. https://doi.org/10.1007/s10549-011-1815-5

24.

Marangoni E, Vincent-Salomon A, Auger N, Degeorges A, Assayag F, de CP, de PL, Guyader C, De PG, Judde JG, Rebucci M, Tran-Perennou C, Sastre-Garau X, Sigal-Zafrani B, Delattre O, Dieras V, Poupon MF (2007) A new model of patient tumor-derived breast cancer xenografts for preclinical assays. Clin Cancer Res 13: 3989-3998. https://doi.org/10.1158/1078-0432.CCR-07-0078

25.

Poulard C, Rambaud J, Le RM, Corbo L. Proximity ligation assay to detect and localize the interactions of ERα with PI3-K and Src in breast cancer cells and tumor samples. Methods Mol Biol. 2014;1204:135–43. https://doi.org/10.1007/978-1-4939-1346-6_12.CrossRefPubMed

26.

Bieche I, Parfait B, Laurendeau I, Girault I, Vidaud M, Lidereau R. Quantification of estrogen receptor α and β expression in sporadic breast cancer. Oncogene. 2001;20:8109–15. https://doi.org/10.1038/sj.onc.1204917.CrossRefPubMed

27.

Marangoni E, Laurent C, Coussy F, El-Botty R, Chateau-Joubert S, Servely JL, de PL, Assayag F, Dahmani A, Montaudon E, Nemati F, Fleury J, Vacher S, Gentien D, Rapinat A, Foidart P, Sounni NE, Noel A, Vincent-Salomon A, Lae M, Decaudin D, Roman-Roman S, Bieche I, Piccart M, Reyal F (2018) Capecitabine efficacy is correlated with TYMP and RB1 expression in PDX established from triple-negative breast cancers. Clin Cancer Res 24: 2605-2615. https://doi.org/10.1158/1078-0432.CCR-17-3490

28.

Tozlu S, Girault I, Vacher S, Vendrell J, Andrieu C, Spyratos F, et al. Identification of novel genes that co-cluster with estrogen receptor alpha in breast tumor biopsy specimens, using a large-scale real-time reverse transcription-PCR approach. Endocr Relat Cancer. 2006;13:1109–20. https://doi.org/10.1677/erc.1.01120.CrossRefPubMed

29.

Fritsch C, Huang A, Chatenay-Rivauday C, Schnell C, Reddy A, Liu M, et al. Characterization of the novel and specific PI3Kα inhibitor NVP-BYL719 and development of the patient stratification strategy for clinical trials. Mol Cancer Ther. 2014;13:1117–29. https://doi.org/10.1158/1535-7163.MCT-13-0865.CrossRefPubMed

30.

Varricchio L, Migliaccio A, Castoria G, Yamaguchi H, De FA, Di DM, et al. Inhibition of estradiol receptor/Src association and cell growth by an estradiol receptor α tyrosine-phosphorylated peptide. Mol Cancer Res. 2007;5:1213–21. https://doi.org/10.1158/1541-7786.MCR-07-0150.CrossRefPubMed

31.

Campbell RA, Bhat-Nakshatri P, Patel NM, Constantinidou D, Ali S, Nakshatri H. Phosphatidylinositol 3-kinase/AKT-mediated activation of estrogen receptor α: a new model for anti-estrogen resistance. J Biol Chem. 2001;276:9817–24. https://doi.org/10.1074/jbc.M010840200.CrossRefPubMed

32.

Marsaud V, Gougelet A, Maillard S, Renoir JM. Various phosphorylation pathways, depending on agonist and antagonist binding to endogenous estrogen receptor α (ERα), differentially affect ERα extractability, proteasome-mediated stability, and transcriptional activity in human breast cancer cells. Mol Endocrinol. 2003;17:2013–27. https://doi.org/10.1210/me.2002-0269.CrossRefPubMed

Title: Analysis of genomic and non-genomic signaling of estrogen receptor in PDX models of breast cancer treated with a combination of the PI3K inhibitor alpelisib (BYL719) and fulvestrant
Authors: Julien Jacquemetton
Loay Kassem
Coralie Poulard
Ahmed Dahmani
Ludmilla De Plater
Elodie Montaudon
Laura Sourd
Ludivine Morisset
Rania El Botty
Sophie Chateau-Joubert
Sophie Vacher
Ivan Bièche
Isabelle Treilleux
Olivier Trédan
Elisabetta Marangoni
Muriel Le Romancer
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Breast Cancer
Breast Cancer
Fulvestrant
Estrogens
Published in: Breast Cancer Research / Issue 1/2021
Electronic ISSN: 1465-542X
DOI: https://doi.org/10.1186/s13058-021-01433-8

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2021

Novel mechanism for OSM-promoted extracellular matrix remodeling in breast cancer: LOXL2 upregulation and subsequent ECM alignment

Mammary collagen architecture and its association with mammographic density and lesion severity among women undergoing image-guided breast biopsy

Correction to: High IRF8 expression correlates with CD8 T cell infiltration and is a predictive biomarker of therapy response in ER-negative breast cancer

Landscape of HER2-low metastatic breast cancer (MBC): results from the Austrian AGMT_MBC-Registry

FGFR1 amplification or overexpression and hormonal resistance in luminal breast cancer: rationale for a triple blockade of ER, CDK4/6, and FGFR1

Association between menopausal hormone therapy, mammographic density and breast cancer risk: results from the E3N cohort study

Keynote webinar | Spotlight on antibody–drug conjugates in cancer