Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

PELP1/SRC-3-dependent regulation of metabolic PFKFB kinases drives therapy resistant ER+ breast cancer

Oncogene volume 40, pages 4384–4397 (2021)Cite this article

Subjects

Abstract

Recurrence of metastatic breast cancer stemming from acquired endocrine and chemotherapy resistance remains a health burden for women with luminal (ER+) breast cancer. Disseminated ER+ tumor cells can remain viable but quiescent for years to decades. Contributing factors to metastatic spread include the maintenance and expansion of breast cancer stem cells (CSCs). Breast CSCs frequently exist as a minority population in therapy resistant tumors. In this study, we show that cytoplasmic complexes composed of steroid receptor (SR) co-activators, PELP1 and SRC-3, modulate breast CSC expansion through upregulation of the HIF-activated metabolic target genes PFKFB3 and PFKFB4. Seahorse metabolic assays demonstrated that cytoplasmic PELP1 influences cellular metabolism by increasing both glycolysis and mitochondrial respiration. PELP1 interacts with PFKFB3 and PFKFB4 proteins, and inhibition of PFKFB3 and PFKFB4 kinase activity blocks PELP1-induced tumorspheres and protein–protein interactions with SRC-3. PFKFB4 knockdown inhibited in vivo emergence of circulating tumor cell (CTC) populations in mammary intraductal (MIND) models. Application of PFKFB inhibitors in combination with ER targeted therapies blocked tumorsphere formation in multiple models of advanced breast cancer including tamoxifen (TamR) and paclitaxel (TaxR) resistant models, murine tumor cells, and ER+ patient-derived organoids (PDxO). Together, our data suggest that PELP1, SRC-3, and PFKFBs cooperate to drive ER+ tumor cell populations that include CSCs and CTCs. Identifying non-ER pharmacological targets offers a useful approach to blocking metastatic escape from standard of care ER/estrogen (E2)-targeted strategies to overcome endocrine and chemotherapy resistance.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: PELP1-induced gene expression is altered in 3D conditions.
Fig. 2: PELP1 cytoplasmic signaling upregulates HIF-activated metabolic pathways.
Fig. 3: PFKFB inhibition blocks PELP1/SRC-3 signaling.
Fig. 4: PFKFB4 knockdown abrogates cyto PELP1 CTCs in MIND xenografts.
Fig. 5: Therapy resistant models phenocopy cyto PELP1 cancer biology.
Fig. 6: Endocrine therapies exhibit combinatorial effects with PELP1 complex inhibitors.
Fig. 7: Co-treatments in preclinical ER+ PDxO models target CSCs.

References

  1. Vadlamudi RK, Wang RA, Mazumdar A, Kim Y, Shin J, Sahin A, et al. Molecular cloning and characterization of PELP1, a novel human coregulator of estrogen receptor alpha. J Biol Chem. 2001;276:38272–9.

    Article  CAS  PubMed  Google Scholar 

  2. Xu J, Liao L, Ning G, Yoshida-Komiya H, Deng C, O’Malley BW. The steroid receptor coactivator SRC-3 (p/CIP/RAC3/AIB1/ACTR/TRAM-1) is required for normal growth, puberty, female reproductive function, and mammary gland development. Proc Natl Acad Sci USA. 2000;97:6379–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Habashy HO, Powe DG, Rakha EA, Ball G, Macmillan RD, Green AR, et al. The prognostic significance of PELP1 expression in invasive breast cancer with emphasis on the ER-positive luminal-like subtype. Breast Cancer Res Treat. 2010;120:603–12.

    Article  CAS  PubMed  Google Scholar 

  4. Kumar R, Zhang H, Holm C, Vadlamudi RK, Landberg G, Rayala SK. Extranuclear coactivator signaling confers insensitivity to tamoxifen. Clin Cancer Res. 2009;15:4123–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Vadlamudi RK, Manavathi B, Balasenthil S, Nair SS, Yang Z, Sahin AA, et al. Functional implications of altered subcellular localization of PELP1 in breast cancer cells. Cancer Res. 2005;65:7724–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Louie MC, Zou JX, Rabinovich A, Chen HW. ACTR/AIB1 functions as an E2F1 coactivator to promote breast cancer cell proliferation and antiestrogen resistance. Mol Cell Biol. 2004;24:5157–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Osborne CK, Bardou V, Hopp TA, Chamness GC, Hilsenbeck SG, Fuqua SA, et al. Role of the estrogen receptor coactivator AIB1 (SRC-3) and HER-2/neu in tamoxifen resistance in breast cancer. J Natl Cancer Inst. 2003;95:353–61.

    Article  CAS  PubMed  Google Scholar 

  8. Burandt E, Jens G, Holst F, Janicke F, Muller V, Quaas A, et al. Prognostic relevance of AIB1 (NCoA3) amplification and overexpression in breast cancer. Breast Cancer Res Treat. 2013;137:745–53.

    Article  CAS  PubMed  Google Scholar 

  9. Song X, Chen J, Zhao M, Zhang C, Yu Y, Lonard DM, et al. Development of potent small-molecule inhibitors to drug the undruggable steroid receptor coactivator-3. Proc Natl Acad Sci USA. 2016;113:4970–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ravindranathan P, Lee TK, Yang L, Centenera MM, Butler L, Tilley WD, et al. Peptidomimetic targeting of critical androgen receptor-coregulator interactions in prostate cancer. Nat Commun. 2013;4:1923.

    Article  PubMed  Google Scholar 

  11. Raj GV, Sareddy GR, Ma S, Lee TK, Viswanadhapalli S, Li R. et al. Estrogen receptor coregulator binding modulators (ERXs) effectively target estrogen receptor positive human breast cancers. Elife. 2017;6:e26857.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Visvader JE, Lindeman GJ. Cancer stem cells: current status and evolving complexities. Cell Stem Cell. 2012;10:717–28.

    Article  CAS  PubMed  Google Scholar 

  13. Rohira AD, Yan F, Wang L, Wang J, Zhou S, Lu A, et al. Targeting SRC coactivators blocks the tumor-initiating capacity of cancer stem-like cells. Cancer Res. 2017;77:4293–304.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Truong TH, Hu H, Temiz NA, Hagen KM, Girard BJ, Brady NJ, et al. Cancer stem cell phenotypes in ER(+) breast cancer models are promoted by PELP1/AIB1 complexes. Mol Cancer Res. 2018;16:707–19.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Girard BJ, Regan Anderson TM, Welch SL, Nicely J, Seewaldt VL, Ostrander JH. Cytoplasmic PELP1 and ERRgamma protect human mammary epithelial cells from Tam-induced cell death. PLoS ONE. 2015;10:e0121206.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Imamura Y, Mukohara T, Shimono Y, Funakoshi Y, Chayahara N, Toyoda M, et al. Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer. Oncol Rep. 2015;33:1837–43.

    Article  CAS  PubMed  Google Scholar 

  17. Tasdemir N, Bossart EA, Li Z, Zhu L, Sikora MJ, Levine KM, et al. Comprehensive phenotypic characterization of human invasive lobular carcinoma cell lines in 2D and 3D cultures. Cancer Res. 2018;78:6209.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Girard BJ, Knutson TP, Kuker B, McDowell L, Schwertfeger KL, Ostrander JH. Cytoplasmic localization of proline, glutamic acid, leucine-rich protein 1 (PELP1) induces breast epithelial cell migration through up-regulation of inhibitor of kappaB kinase and inflammatory cross-talk with macrophages. J Biol Chem. 2017;292:339–50.

    Article  CAS  PubMed  Google Scholar 

  19. Liu Y, Nenutil R, Appleyard MV, Murray K, Boylan M, Thompson AM, et al. Lack of correlation of stem cell markers in breast cancer stem cells. Br J Cancer. 2014;110:2063–71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Chesney J, Clark J, Klarer AC, Imbert-Fernandez Y, Lane AN, Telang S. Fructose-2,6-bisphosphate synthesis by 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4) is required for the glycolytic response to hypoxia and tumor growth. Oncotarget. 2014;5:6670–86.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Kim JW, Dang CV. Cancer’s molecular sweet tooth and the Warburg effect. Cancer Res. 2006;66:8927–30.

    Article  CAS  PubMed  Google Scholar 

  22. Shi L, Pan H, Liu Z, Xie J, Han W. Roles of PFKFB3 in cancer. Signal Transduct Target Ther. 2017;2:17044.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Torres-Arzayus MI, Font de Mora J, Yuan J, Vazquez F, Bronson R, Rue M, et al. High tumor incidence and activation of the PI3K/AKT pathway in transgenic mice define AIB1 as an oncogene. Cancer Cell. 2004;6:263–74.

    Article  CAS  PubMed  Google Scholar 

  24. Sflomos G, Dormoy V, Metsalu T, Jeitziner R, Battista L, Scabia V, et al. A preclinical model for ERalpha-positive breast cancer points to the epithelial microenvironment as determinant of luminal phenotype and hormone response. Cancer Cell. 2016;29:407–22.

    Article  CAS  PubMed  Google Scholar 

  25. Kilgour E, Rothwell DG, Brady G, Dive C. Liquid biopsy-based biomarkers of treatment response and resistance. Cancer Cell. 2020;37:485–95.

    Article  CAS  PubMed  Google Scholar 

  26. Regan Anderson TM, Ma S, Perez Kerkvliet C, Peng Y, Helle TM, Krutilina RI, et al. Taxol induces Brk-dependent prosurvival phenotypes in TNBC cells through an AhR/GR/HIF-driven signaling axis. Mol Cancer Res. 2018;16:1761–72.

    Article  PubMed  Google Scholar 

  27. Torres-Arzayus MI, Yuan J, DellaGatta JL, Lane H, Kung AL, Brown M. Targeting the AIB1 oncogene through mammalian target of rapamycin inhibition in the mammary gland. Cancer Res. 2006;66:11381–8.

    Article  CAS  PubMed  Google Scholar 

  28. Guillen KP, Fujita M, Butterfield AJ, Scherer SD, Bailey MH, Chu Z, et al. A breast cancer patient-derived xenograft and organoid platform for drug discovery and precision oncology. bioRxiv. 2021. https://doi.org/10.1101/2021.02.28.433268.

  29. Moore N, Lyle S. Quiescent, slow-cycling stem cell populations in cancer: a review of the evidence and discussion of significance. J Oncol. 2011;2011:396076.

    Article  PubMed  Google Scholar 

  30. Bosco DB, Kenworthy R, Zorio DA, Sang QX. Human mesenchymal stem cells are resistant to Paclitaxel by adopting a non-proliferative fibroblastic state. PLoS ONE. 2015;10:e0128511.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Wang X, Pan L, Mao N, Sun L, Qin X, Yin J. Cell-cycle synchronization reverses Taxol resistance of human ovarian cancer cell lines. Cancer Cell Int. 2013;13:77.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Nair BC, Nair SS, Chakravarty D, Challa R, Manavathi B, Yew PR, et al. Cyclin-dependent kinase-mediated phosphorylation plays a critical role in the oncogenic functions of PELP1. Cancer Res. 2010;70:7166–75.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. O’Neill S, Porter RK, McNamee N, Martinez VG, O’Driscoll L. 2-Deoxy-D-Glucose inhibits aggressive triple-negative breast cancer cells by targeting glycolysis and the cancer stem cell phenotype. Sci Rep. 2019;9:3788.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Dong C, Yuan T, Wu Y, Wang Y, Fan TW, Miriyala S, et al. Loss of FBP1 by Snail-mediated repression provides metabolic advantages in basal-like breast cancer. Cancer Cell. 2013;23:316–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Vlashi E, Lagadec C, Vergnes L, Reue K, Frohnen P, Chan M, et al. Metabolic differences in breast cancer stem cells and differentiated progeny. Breast Cancer Res Treat. 2014;146:525–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Banerjee A, Arvinrad P, Darley M, Laversin SA, Parker R, Rose-Zerilli MJJ, et al. The effects of restricted glycolysis on stem-cell like characteristics of breast cancer cells. Oncotarget. 2018;9:23274–88.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Regan Anderson TM, Ma SH, Raj GV, Cidlowski JA, Helle TM, Knutson TP, et al. Breast tumor kinase (Brk/PTK6) is induced by HIF, glucocorticoid receptor, and PELP1-mediated stress signaling in triple-negative breast cancer. Cancer Res. 2016;76:1653–63.

    Article  CAS  PubMed  Google Scholar 

  38. Girard BJ, Daniel AR, Lange CA, Ostrander JH. PELP1: a review of PELP1 interactions, signaling, and biology. Mol Cell Endocrinol. 2014;382:642–51.

    Article  CAS  PubMed  Google Scholar 

  39. Wu RC, Qin J, Yi P, Wong J, Tsai SY, Tsai MJ, et al. Selective phosphorylations of the SRC-3/AIB1 coactivator integrate genomic reponses to multiple cellular signaling pathways. Mol Cell. 2004;15:937–49.

    Article  CAS  PubMed  Google Scholar 

  40. Dasgupta S, Rajapakshe K, Zhu B, Nikolai BC, Yi P, Putluri N, et al. Metabolic enzyme PFKFB4 activates transcriptional coactivator SRC-3 to drive breast cancer. Nature. 2018;556:249–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Mondal S, Roy D, Sarkar Bhattacharya S, Jin L, Jung D, Zhang S, et al. Therapeutic targeting of PFKFB3 with a novel glycolytic inhibitor PFK158 promotes lipophagy and chemosensitivity in gynecologic cancers. Int J Cancer. 2019;144:178–89.

    Article  CAS  PubMed  Google Scholar 

  42. Telang S, Yaddanapudi K, Grewal J, Redman R, Fu S, Pohlmann P, et al. Abstract B90: PFK-158 is a first-in-human inhibitor of PFKFB3 that selectively suppresses glucose metabolism of cancer cells and inhibits the immunosuppressive Th17 cells and MDSCs in advanced cancer patients. Cancer Res. 2016;76:B90.

    Article  Google Scholar 

  43. Redman R, Pohlmann P, Kurman M, Tapolsky GH, Chesney J. Abstract CT206: PFK-158, first-in-man and first-in-class inhibitor of PFKFB3/ glycolysis: a phase I, dose escalation, multi-center study in patients with advanced solid malignancies. Cancer Res. 2015;75:CT206.

    Article  Google Scholar 

  44. Yao L, Wang L, Cao ZG, Hu X, Shao ZM. High expression of metabolic enzyme PFKFB4 is associated with poor prognosis of operable breast cancer. Cancer Cell Int. 2019;19:165.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Marotta LL, Almendro V, Marusyk A, Shipitsin M, Schemme J, Walker SR, et al. The JAK2/STAT3 signaling pathway is required for growth of CD44(+)CD24(−) stem cell-like breast cancer cells in human tumors. J Clin Invest. 2011;121:2723–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Gao R, Li D, Xun J, Zhou W, Li J, Wang J, et al. CD44ICD promotes breast cancer stemness via PFKFB4-mediated glucose metabolism. Theranostics. 2018;8:6248–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Giordano A, Gao H, Anfossi S, Cohen E, Mego M, Lee BN, et al. Epithelial-mesenchymal transition and stem cell markers in patients with HER2-positive metastatic breast cancer. Mol Cancer Ther. 2012;11:2526–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Behbod F, Kittrell FS, LaMarca H, Edwards D, Kerbawy S, Heestand JC, et al. An intraductal human-in-mouse transplantation model mimics the subtypes of ductal carcinoma in situ. Breast Cancer Res. 2009;11:R66.

    Article  PubMed  PubMed Central  Google Scholar 

  49. Yang Y, Chan JY, Temiz NA, Yee D. Insulin receptor substrate suppression by the tyrphostin NT157 inhibits responses to insulin-like growth factor-I and insulin in breast cancer cells. Horm Cancer. 2018;9:371–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Dwyer AR, Truong TH, Kerkvliet CP, Paul KV, Kabos P, Sartorius CA, et al. Insulin receptor substrate-1 (IRS-1) mediates progesterone receptor-driven stemness and endocrine resistance in oestrogen receptor+ breast cancer. Br J Cancer. 2021;124:217–27.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by NIH grants R01 CA236948 (JHO, CAL), R01 CA229697 (CAL), F32 CA210340 (THT), T32 HL007741 (THT), U54 CA224076 (BEW), R01 CA248158-01 (CODS), and R01 AG069727-01 (CODS). ACS Institutional Research Grant #124166-IRG-58-001-52-IRG5 (JHO), University of Minnesota Masonic Cancer Center (CAL, JHO), the Tickle Family Land Grant Endowed Chair in Breast Cancer Research (CAL), National Center for Advancing Translational Sciences of the NIH Award UL1TR000114 (JHO), and Department of Defense W81XWH-14-1-0417 (BEW). We thank Bruce Lindgren for biostatistics support, and the Masonic Cancer Center Biostatistics and Bioinformatics, Analytical Biochemistry, University Imaging Core (UIC), and Flow Cytometry cores. We also thank Zohar Sachs and Michael Franklin for critical reading of this manuscript.

Author information

Author notes

  1. These authors contributed equally: Carol A. Lange, Julie H. Ostrander

Authors and Affiliations

  1. Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA

    Thu H. Truong, Elizabeth A. Benner, Kyla M. Hagen, Nuri A. Temiz, Carlos Perez Kerkvliet, Ying Wang, Carol A. Lange & Julie H. Ostrander

  2. Institute for Health Informatics, University of Minnesota, Minneapolis, MN, USA

    Nuri A. Temiz

  3. Department of Oncological Sciences, University of Utah, Salt Lake City, UT, USA

    Emilio Cortes-Sanchez, Chieh-Hsiang Yang, Katrin P. Guillen & Bryan E. Welm

  4. Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, USA

    Emilio Cortes-Sanchez, Chieh-Hsiang Yang, Katrin P. Guillen & Bryan E. Welm

  5. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA

    Marygrace C. Trousdell & Camila O. Dos Santos

  6. University of Minnesota Informatics Institute, University of Minnesota, Minneapolis, MN, USA

    Thomas Pengo

  7. Department of Surgery, University of Utah, Salt Lake City, UT, USA

    Bryan E. Welm

  8. James Graham Brown Cancer Center, Department of Medicine (Division of Medical Oncology and Hematology), University of Louisville, Louisville, KY, USA

    Sucheta Telang

  9. Department of Medicine (Division of Hematology, Oncology, and Transplantation), University of Minnesota, Minneapolis, MN, USA

    Carol A. Lange & Julie H. Ostrander

  10. Department of Pharmacology, University of Minnesota, Minneapolis, MN, USA

    Carol A. Lange

Authors
  1. Thu H. Truong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elizabeth A. Benner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kyla M. Hagen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nuri A. Temiz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carlos Perez Kerkvliet
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ying Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Emilio Cortes-Sanchez
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Chieh-Hsiang Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Marygrace C. Trousdell
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Thomas Pengo
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Katrin P. Guillen
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Bryan E. Welm
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Camila O. Dos Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Sucheta Telang
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Carol A. Lange
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Julie H. Ostrander
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Carol A. Lange or Julie H. Ostrander.

Ethics declarations

Conflict of interest

CAL is a Scientific Advisory Board Member for Context Therapeutics, Inc. BEW, EC-S, KPG, and C-HY may receive financial compensation from intellectual property and tangible property licenses managed by the University of Utah. The remaining authors have nothing to disclose.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Truong, T.H., Benner, E.A., Hagen, K.M. et al. PELP1/SRC-3-dependent regulation of metabolic PFKFB kinases drives therapy resistant ER+ breast cancer. Oncogene 40, 4384–4397 (2021). https://doi.org/10.1038/s41388-021-01871-w

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-021-01871-w

This article is cited by

Search

Advanced search

Quick links