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.

  • Article
  • Published:

Elevated leptin disrupts epithelial polarity and promotes premalignant alterations in the mammary gland

Oncogene volume 38, pages 3855–3870 (2019)Cite this article

Subjects

Abstract

Obesity is a highly prevalent and modifiable breast cancer risk factor. While the role of obesity in fueling breast cancer progression is well established, the mechanisms linking obesity to breast cancer initiation are poorly understood. A hallmark of breast cancer initiation is the disruption of apical polarity in mammary glands. Here we show that mice with diet-induced obesity display mislocalization of Par3, a regulator of cellular junctional complexes defining mammary epithelial polarity. We found that epithelial polarity loss also occurs in a 3D coculture system that combines acini with human mammary adipose tissue, and establish that a paracrine effect of the tissue adipokine leptin causes loss of polarity by overactivation of the PI3K/Akt pathway. Leptin sensitizes non-neoplastic cells to proliferative stimuli, causes mitotic spindle misalignment, and expands the pool of cells with stem/progenitor characteristics, which are early steps for cancer initiation. We also found that normal breast tissue samples with high leptin/adiponectin transcript ratio characteristic of obesity have an altered distribution of apical polarity markers. This effect is associated with increased epithelial cell layers. Our results provide a molecular basis for early alterations in epithelial architecture during obesity-mediated cancer initiation.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Calle EE, Rodriguez C, Walker-Thurmond K, Thun MJ. Overweight, obesity, and mortality from cancer in a prospectively studied cohort of U.S. adults. N Eng J Med. 2003;348:1625–38.

    Article  Google Scholar 

  2. Renehan AG, Zwahlen M, Egger M. Adiposity and cancer risk: new mechanistic insights from epidemiology. Nat Rev Cancer. 2015;15:484–98.

    Article  CAS  Google Scholar 

  3. Schwartz GF, Hughes KS, Lynch HT, Fabian CJ, Fentiman IS, Robson ME, et al. Proceedings of the international consensus conference on breast cancer risk, genetics, & risk management, April, 2007. Cancer. 2008;113:2627–37.

    Article  Google Scholar 

  4. Cecchini RS, Costantino JP, Cauley JA, Cronin WM, Wickerham DL, Land SR, et al. Body mass index and the risk for developing invasive breast cancer among high-risk women in NSABP P-1 and STAR breast cancer prevention trials. Cancer Prev Res. 2012;5:583–92.

    Article  CAS  Google Scholar 

  5. Pierobon M, Frankenfeld CL. Obesity as a risk factor for triple-negative breast cancers: a systematic review and meta-analysis. Breast Cancer Res Treat. 2013;137:307–14.

    Article  Google Scholar 

  6. Ando S, Catalano S. The multifactorial role of leptin in driving the breast cancer microenvironment. Nat Rev Endocrinol. 2012;8:263–75.

    Article  CAS  Google Scholar 

  7. Saxena NK, Sharma D. Multifaceted leptin network: the molecular connection between obesity and breast cancer. J Mammary Gland Biol Neoplasia. 2013;18:309–20.

    Article  Google Scholar 

  8. Considine RV, Sinha MK, Heiman ML, Kriauciunas A, Stephens TW, Nyce MR, et al. Serum immunoreactive-leptin concentrations in normal-weight and obese humans. N Eng J Med. 1996;334:292–5.

    Article  CAS  Google Scholar 

  9. Matsubara M, Maruoka S, Katayose S. Inverse relationship between plasma adiponectin and leptin concentrations in normal-weight and obese women. Eur J Endocrinol. 2002;147:173–80.

    Article  CAS  Google Scholar 

  10. Wheatley KE, Nogueira LM, Perkins SN, Hursting SD. Differential effects of calorie restriction and exercise on the adipose transcriptome in diet-induced obese mice. J Obes. 2011;2011:265417.

    Article  Google Scholar 

  11. Niu J, Jiang L, Guo W, Shao L, Liu Y, Wang L. The association between leptin level and breast cancer: a meta-analysis. PLoS One. 2013;8:e67349.

    Article  CAS  Google Scholar 

  12. Wu MH, Chou YC, Chou WY, Hsu GC, Chu CH, Yu CP, et al. Circulating levels of leptin, adiposity and breast cancer risk. Br J Cancer. 2009;100:578–82.

    Article  CAS  Google Scholar 

  13. Lelievre SA. Tissue polarity-dependent control of mammary epithelial homeostasis and cancer development: an epigenetic perspective. J Mammary Gland Biol Neoplasia. 2010;15:49–63.

    Article  Google Scholar 

  14. Royer C, Lu X. Epithelial cell polarity: a major gatekeeper against cancer? Cell Death Differ. 2011;18:1470–7.

    Article  CAS  Google Scholar 

  15. Martin-Belmonte F, Perez-Moreno M. Epithelial cell polarity, stem cells and cancer. Nat Rev Cancer. 2012;12:23–38.

    Article  CAS  Google Scholar 

  16. Balda MS, Matter K. The tight junction protein ZO-1 and an interacting transcription factor regulate ErbB-2 expression. EMBO J. 2000;19:2024–33.

    Article  CAS  Google Scholar 

  17. Feigin ME, Akshinthala SD, Araki K, Rosenberg AZ, Muthuswamy LB, Martin B, et al. Mislocalization of the cell polarity protein scribble promotes mammary tumorigenesis and is associated with basal breast cancer. Cancer Res. 2014;74:3180–94.

    Article  CAS  Google Scholar 

  18. Fang L, Wang Y, Du D, Yang G, Tak Kwok T, Kai Kong S, et al. Cell polarity protein Par3 complexes with DNA-PK via Ku70 and regulates DNA double-strand break repair. Cell Res. 2007;17:100–16.

    Article  CAS  Google Scholar 

  19. Chandramouly G, Abad PC, Knowles DW, Lelievre SA. The control of tissue architecture over nuclear organization is crucial for epithelial cell fate. J Cell Sci. 2007;120:1596–606.

    Article  CAS  Google Scholar 

  20. Vidi PA, Chandramouly G, Gray M, Wang L, Liu E, Kim JJ, et al. Interconnected contribution of tissue morphogenesis and the nuclear protein NuMA to the DNA damage response. J Cell Sci. 2012;125:350–61.

    Article  CAS  Google Scholar 

  21. Sumis A, Cook KL, Andrade FO, Hu R, Kidney E, Zhang X, et al. Social isolation induces autophagy in the mouse mammary gland: link to increased mammary cancer risk. Endocr Relat Cancer. 2016;23:839–56.

    Article  CAS  Google Scholar 

  22. Roignot J, Peng X, Mostov K. Polarity in mammalian epithelial morphogenesis. Cold Spring Harbor Perspect Biol. 2013;5:a013789.

    Article  Google Scholar 

  23. McCaffrey LM, Montalbano J, Mihai C, Macara IG. Loss of the Par3 polarity protein promotes breast tumorigenesis and metastasis. Cancer Cell. 2012;22:601–14.

    Article  CAS  Google Scholar 

  24. Petersen OW, Ronnov-Jessen L, Howlett AR, Bissell MJ. Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci USA. 1992;89:9064–8.

    Article  CAS  Google Scholar 

  25. Vidi PA, Bissell MJ, Lelievre SA. Three-dimensional culture of human breast epithelial cells: the how and the why. Methods Mol Biol. 2013;945:193–219.

    Article  Google Scholar 

  26. Ibarra-Drendall C, Troch MM, Barry WT, Broadwater G, Petricoin EF 3rd, Wulfkuhle J, et al. Pilot and feasibility study: prospective proteomic profiling of mammary epithelial cells from high-risk women provides evidence of activation of pro-survival pathways. Breast Cancer Res Treat. 2012;132:487–98.

    Article  CAS  Google Scholar 

  27. Gunter MJ, Hoover DR, Yu H, Wassertheil-Smoller S, Rohan TE, Manson JE, et al. Insulin, insulin-like growth factor-I, and risk of breast cancer in postmenopausal women. J Natl Cancer Inst. 2009;101:48–60.

    Article  CAS  Google Scholar 

  28. Kim CS, Park HS, Kawada T, Kim JH, Lim D, Hubbard NE, et al. Circulating levels of MCP-1 and IL-8 are elevated in human obese subjects and associated with obesity-related parameters. Int J Obes. 2006;30:1347–55.

    Article  CAS  Google Scholar 

  29. Carpenter RL, Paw I, Dewhirst MW, Lo HW. Akt phosphorylates and activates HSF-1 independent of heat shock, leading to Slug overexpression and epithelial-mesenchymal transition (EMT) of HER2-overexpressing breast cancer cells. Oncogene. 2015;34:546–57.

    Article  CAS  Google Scholar 

  30. Elloul S, Kedrin D, Knoblauch NW, Beck AH, Toker A. The adherens junction protein afadin is an AKT substrate that regulates breast cancer cell migration. Mol Cancer Res. 2014;12:464–76.

    Article  CAS  Google Scholar 

  31. Takai Y, Ikeda W, Ogita H, Rikitake Y. The immunoglobulin-like cell adhesion molecule nectin and its associated protein afadin. Annu Rev Cell Dev Biol. 2008;24:309–42.

    Article  CAS  Google Scholar 

  32. Ooshio T, Kobayashi R, Ikeda W, Miyata M, Fukumoto Y, Matsuzawa N, et al. Involvement of the interaction of afadin with ZO-1 in the formation of tight junctions in Madin-Darby canine kidney cells. J Biol Chem. 2010;285:5003–12.

    Article  CAS  Google Scholar 

  33. Dubois V, Jarde T, Delort L, Billard H, Bernard-Gallon D, Berger E, et al. Leptin induces a proliferative response in breast cancer cells but not in normal breast cells. Nutr Cancer. 2014;66:645–55.

    Article  CAS  Google Scholar 

  34. Ray A, Cleary MP. The potential role of leptin in tumor invasion and metastasis. Cytokine Growth Factor Rev. 2017;38:80–97.

    Article  CAS  Google Scholar 

  35. Macara IG, Guyer R, Richardson G, Huo Y, Ahmed SM. Epithelial homeostasis. Curr Biol. 2014;24:R815–25.

    Article  CAS  Google Scholar 

  36. Hao Y, Du Q, Chen X, Zheng Z, Balsbaugh JL, Maitra S, et al. Par3 controls epithelial spindle orientation by aPKC-mediated phosphorylation of apical Pins. Curr Biol. 2010;20:1809–18.

    Article  CAS  Google Scholar 

  37. Carminati M, Gallini S, Pirovano L, Alfieri A, Bisi S, Mapelli M. Concomitant binding of Afadin to LGN and F-actin directs planar spindle orientation. Nat Struct Mol Biol. 2016;23:155–63.

    Article  CAS  Google Scholar 

  38. Grafton MM, Wang L, Vidi PA, Leary J, Lelievre SA. Breast on-a-chip: mimicry of the channeling system of the breast for development of theranostics. Integr Biol. 2011;3:451–9.

    Article  CAS  Google Scholar 

  39. Esper RM, Dame M, McClintock S, Holt PR, Dannenberg AJ, Wicha MS, et al. Leptin and adiponectin modulate the self-renewal of normal human breast epithelialstem cells. Cancer Prev Res. 2015;8:1174–83.

    Article  CAS  Google Scholar 

  40. Hursting SD, Digiovanni J, Dannenberg AJ, Azrad M, Leroith D, Demark-Wahnefried W, et al. Obesity, energy balance, and cancer: new opportunities for prevention. Cancer Prev Res. 2012;5:1260–72.

    Article  CAS  Google Scholar 

  41. Grossmann ME, Cleary MP. The balance between leptin and adiponectin in the control of carcinogenesis - focus on mammary tumorigenesis. Biochimie. 2012;94:2164–71.

    Article  CAS  Google Scholar 

  42. Macis D, Guerrieri-Gonzaga A, Gandini S. Circulating adiponectin and breast cancer risk: a systematic review and meta-analysis. Int J Epidemiol. 2014;43:1226–36.

    Article  Google Scholar 

  43. Fan Y, Liu Y, Xue K, Gu G, Fan W, Xu Y. et al. Diet-induced obesity in male C57BL/6 mice decreases fertility as a consequence of disrupted blood-testis barrier. PLoS ONE. 2015;10:e0120775

    Article  Google Scholar 

  44. Teixeira TF, Collado MC, Ferreira CL, Bressan J, Peluzio Mdo C. Potential mechanisms for the emerging link between obesity and increased intestinal permeability. Nutr Res. 2012;32:637–47.

    Article  CAS  Google Scholar 

  45. Bartella V, Cascio S, Fiorio E, Auriemma A, Russo A, Surmacz E. Insulin-dependent leptin expression in breast cancer cells. Cancer Res. 2008;68:4919–27.

    Article  CAS  Google Scholar 

  46. Bissell MJ, Weaver VM, Lelievre SA, Wang F, Petersen OW, Schmeichel KL. Tissue structure, nuclear organization, and gene expression in normal and malignant breast. Cancer Res. 1999;59:1757–1763s.

    CAS  PubMed  Google Scholar 

  47. Hu X, Juneja SC, Maihle NJ, Cleary MP. Leptin--a growth factor in normal and malignant breast cells and for normal mammary gland development. J Natl Cancer Inst. 2002;94:1704–11.

    Article  CAS  Google Scholar 

  48. Martin-Belmonte F, Gassama A, Datta A, Yu W, Rescher U, Gerke V, et al. PTEN-mediated apical segregation of phosphoinositides controls epithelial morphogenesis through Cdc42. Cell. 2007;128:383–97.

    Article  CAS  Google Scholar 

  49. Rangel MC, Bertolette D, Castro NP, Klauzinska M, Cuttitta F, Salomon DS. Developmental signaling pathways regulating mammary stem cells and contributing to the etiology of triple-negative breast cancer. Breast Cancer Res Treat. 2016;156:211–26.

    Article  Google Scholar 

  50. Wang F, Hansen RK, Radisky D, Yoneda T, Barcellos-Hoff MH, Petersen OW, et al. Phenotypic reversion or death of cancer cells by altering signaling pathways in three-dimensional contexts. J Natl Cancer Inst. 2002;94:1494–503.

    Article  CAS  Google Scholar 

  51. Bakin AV, Tomlinson AK, Bhowmick NA, Moses HL, Arteaga CL. Phosphatidylinositol 3-kinase function is required for transforming growth factor beta-mediated epithelial to mesenchymal transition and cell migration. J Biol Chem. 2000;275:36803–10.

    Article  CAS  Google Scholar 

  52. Xue G, Hemmings BA. PKB/Akt-dependent regulation of cell motility. J Natl Cancer Inst. 2013;105:393–404.

    Article  CAS  Google Scholar 

  53. Zeidan A, Paylor B, Steinhoff KJ, Javadov S, Rajapurohitam V, Chakrabarti S, et al. Actin cytoskeleton dynamics promotes leptin-induced vascular smooth muscle hypertrophy via RhoA/ROCK- and phosphatidylinositol 3-kinase/protein kinase B-dependent pathways. J Pharmacol Exp Ther. 2007;322:1110–6.

    Article  CAS  Google Scholar 

  54. Millikan RC, Newman B, Tse CK, Moorman PG, Conway K, Dressler LG, et al. Epidemiology of basal-like breast cancer. Breast Cancer Res Treat. 2008;109:123–39.

    Article  Google Scholar 

  55. Dietze EC, Sistrunk C, Miranda-Carboni G, O’Regan R, Seewaldt VL. Triple-negative breast cancer in African-American women: disparities versus biology. Nat Rev Cancer. 2015;15:248–54.

    Article  CAS  Google Scholar 

  56. Ogden CL, Carroll MD, Fryar CD, Flegal KM. Prevalence of obesity among adults and youth: United States, 2011–2014. NCHS Data Brief. 2015;219:1–8.

    Google Scholar 

  57. Briand P, Petersen OW, Van Deurs B. A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium. In Vitro Cell Dev Biol. 1987;23:181–8.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Kenneth Grant for assistance with electron microscopy and Dr. Fei Xing for assistance with stem cell analyses. The CA-Akt construct was a gift from Dr. Mong-Hong Lee (MD Anderson Cancer Center). NuMA B1C11 antibodies were kindly provided by Dr. J. Nickerson (University of Massachusetts). This work was funded by the National Institute of Health (R00CA163957 to PAV), the Walther Cancer Foundation (to SAL, PAV, and IGC), the Wake Forest Center for Molecular Signaling (CMS; to PAV and KB), and the National Cancer Institute’s Cancer Center Support Grant award number P30CA012197 issued to the Wake Forest Baptist Comprehensive Cancer Center. SAL, VS, and PAV are members of the International Breast Cancer & Nutrition (IBCN) program.

Author information

Author notes

  1. These authors contributed equally: Iliana Tenvooren, Mónica Z. Jenks

Authors and Affiliations

  1. Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC, 27157, USA

    Iliana Tenvooren, Mónica Z. Jenks, Hamza Rashid, Julia Holmes, Hui-Wen Lo & Pierre-Alexandre Vidi

  2. Department of Surgery, Wake Forest School of Medicine, Winston-Salem, NC, 27157, USA

    Katherine L. Cook

  3. Comprehensive Cancer Center of Wake Forest University, Winston-Salem, NC, USA

    Katherine L. Cook & Pierre-Alexandre Vidi

  4. Department of Biology, Wake Forest University, Winston-Salem, NC, 27109, USA

    Joëlle K. Muhlemann

  5. Department of Population Sciences, City of Hope, Duarte, CA, 91010, USA

    Christopher Sistrunk & Victoria Seewaldt

  6. Department of Physics, Wake Forest University, Winston-Salem, NC, 27109, USA

    Kevin Wang & Keith Bonin

  7. Department of Pathology and Laboratory Medicine, University of Cincinnati, Cincinnati, OH, 45219, USA

    Kurt Hodges

  8. Indiana University School of Medicine, IU Health Arnett Hospital, Lafayette, IN, 47905, USA

    Ayaz Shaikh

  9. Department of Biological Sciences, Purdue University, West Lafayette, IN, 47907, USA

    Ignacio G. Camarillo

  10. Department of Basic Medical Sciences, Purdue University, West Lafayette, IN, 47907, USA

    Sophie A. Lelièvre

Authors
  1. Iliana Tenvooren
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mónica Z. Jenks
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hamza Rashid
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Katherine L. Cook
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Joëlle K. Muhlemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christopher Sistrunk
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Julia Holmes
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kevin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Keith Bonin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Kurt Hodges
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hui-Wen Lo
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Ayaz Shaikh
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Ignacio G. Camarillo
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Sophie A. Lelièvre
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Victoria Seewaldt
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Pierre-Alexandre Vidi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pierre-Alexandre Vidi.

Ethics declarations

Conflict of interest

PAV and KB have filed a provisional patent application (No. 62/672,951) on the use of the radial profile method to measure epithelial polarity. The remaining authors declare that they have no conflict of interest.

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

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tenvooren, I., Jenks, M.Z., Rashid, H. et al. Elevated leptin disrupts epithelial polarity and promotes premalignant alterations in the mammary gland. Oncogene 38, 3855–3870 (2019). https://doi.org/10.1038/s41388-019-0687-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-019-0687-8

This article is cited by

Search

Advanced search

Quick links