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P-REX1 creates a positive feedback loop to activate growth factor receptor, PI3K/AKT and MEK/ERK signaling in breast cancer

Oncogene volume 34, pages 3968–3976 (2015)Cite this article

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Abstract

Phosphatidylinositol 3-kinase (PI3K) promotes cancer cell survival, migration, growth and proliferation by generating phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the inner leaflet of the plasma membrane. PIP3 recruits pleckstrin homology domain-containing proteins to the membrane to activate oncogenic signaling cascades. Anticancer therapeutics targeting the PI3K/AKT/mTOR (mammalian target of rapamycin) pathway are in clinical development. In a mass spectrometric screen to identify PIP3-regulated proteins in breast cancer cells, levels of the Rac activator PIP3-dependent Rac exchange factor-1 (P-REX1) increased in response to PI3K inhibition, and decreased upon loss of the PI3K antagonist phosphatase and tensin homolog (PTEN). P-REX1 mRNA and protein levels were positively correlated with ER expression, and inversely correlated with PI3K pathway activation in breast tumors as assessed by gene expression and phosphoproteomic analyses. P-REX1 increased activation of Rac1, PI3K/AKT and MEK/ERK signaling in a PTEN-independent manner, and promoted cell and tumor viability. Loss of P-REX1 or inhibition of Rac suppressed PI3K/AKT and MEK/ERK, and decreased viability. P-REX1 also promoted insulin-like growth factor-1 receptor activation, suggesting that P-REX1 provides positive feedback to activators upstream of PI3K. In support of a model where PIP3-driven P-REX1 promotes both PI3K/AKT and MEK/ERK signaling, high levels of P-REX1 mRNA (but not phospho-AKT or a transcriptomic signature of PI3K activation) were predictive of sensitivity to PI3K inhibitors among breast cancer cell lines. P-REX1 expression was highest in estrogen receptor-positive breast tumors compared with many other cancer subtypes, suggesting that neutralizing the P-REX1/Rac axis may provide a novel therapeutic approach to selectively abrogate oncogenic signaling in breast cancer cells.

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References

  1. Miller TW, Rexer BN, Garrett JT, Arteaga CL . Mutations in the phosphatidylinositol 3-kinase pathway: role in tumor progression and therapeutic implications in breast cancer. Breast Cancer Res 2011; 13: 224.

    Article  CAS  Google Scholar 

  2. Engelman JA . Targeting PI3K signalling in cancer: opportunities, challenges and limitations. Nat Rev Cancer 2009; 9: 550–562.

    Article  CAS  Google Scholar 

  3. Whitman M, Downes CP, Keeler M, Keller T, Cantley L . Type I phosphatidylinositol kinase makes a novel inositol phospholipid, phosphatidylinositol-3-phosphate. Nature 1988; 332: 644–646.

    Article  CAS  Google Scholar 

  4. Maehama T, Dixon JE . The tumor suppressor, PTEN/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5-trisphosphate. J Biol Chem 1998; 273: 13375–13378.

    Article  CAS  Google Scholar 

  5. Maira SM, Pecchi S, Huang A, Burger M, Knapp M, Sterker D et al. Identification and characterization of NVP-BKM120, an orally available pan-class I PI3-kinase inhibitor. Mol Cancer Ther 2012; 11: 317–328.

    Article  CAS  Google Scholar 

  6. Welch HC, Coadwell WJ, Ellson CD, Ferguson GJ, Andrews SR, Erdjument-Bromage H et al. P-Rex1, a PtdIns(3,4,5)P3- and Gbetagamma-regulated guanine-nucleotide exchange factor for Rac. Cell 2002; 108: 809–821.

    Article  CAS  Google Scholar 

  7. Qin J, Xie Y, Wang B, Hoshino M, Wolff DW, Zhao J et al. Upregulation of PIP3-dependent Rac exchanger 1 (P-Rex1) promotes prostate cancer metastasis. Oncogene 2009; 28: 1853–1863.

    Article  CAS  Google Scholar 

  8. Montero JC, Seoane S, Ocana A, Pandiella A . P-Rex1 participates in neuregulin-ErbB signal transduction and its expression correlates with patient outcome in breast cancer. Oncogene 2011; 30: 1059–1071.

    Article  CAS  Google Scholar 

  9. Sosa MS, Lopez-Haber C, Yang C, Wang H, Lemmon MA, Busillo JM et al. Identification of the Rac-GEF P-Rex1 as an essential mediator of ErbB signaling in breast cancer. Mol Cell 2010; 40: 877–892.

    Article  CAS  Google Scholar 

  10. Fine B, Hodakoski C, Koujak S, Su T, Saal LH, Maurer M et al. Activation of the PI3K pathway in cancer through inhibition of PTEN by exchange factor P-REX2a. Science 2009; 325: 1261–1265.

    Article  CAS  Google Scholar 

  11. Folkes AJ, Ahmadi K, Alderton WK, Alix S, Baker SJ, Box G et al. The identification of 2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine (GDC-0941) as a potent, selective, orally bioavailable inhibitor of class I PI3 kinase for the treatment of cancer. J Med Chem 2008; 51: 5522–5532.

    Article  CAS  Google Scholar 

  12. Miller TW, Perez-Torres M, Narasanna A, Guix M, Stal O, Perez-Tenorio G et al. Loss of phosphatase and tensin homologue deleted on chromosome 10 engages ErbB3 and insulin-like growth factor-I receptor signaling to promote antiestrogen resistance in breast cancer. Cancer Res 2009; 69: 4192–4201.

    Article  CAS  Google Scholar 

  13. Chakrabarty A, Sanchez V, Kuba MG, Rinehart C, Arteaga CL . Feedback upregulation of HER3 (ErbB3) expression and activity attenuates antitumor effect of PI3K inhibitors. Proc Natl Acad Sci USA 2011; 109: 2718–2723.

    Article  Google Scholar 

  14. Khatri S, Yepiskoposyan H, Gallo CA, Tandon P, Plas DR . FOXO3a regulates glycolysis via transcriptional control of tumor suppressor TSC1. J Biol Chem 2010; 285: 15960–15965.

    Article  CAS  Google Scholar 

  15. Creighton CJ, Fu X, Hennessy BT, Casa AJ, Zhang Y, Gonzalez-Angulo AM et al. Proteomic and transcriptomic profiling reveals a link between the PI3K pathway and lower estrogen-receptor (ER) levels and activity in ER+ breast cancer. Breast Cancer Res 2010; 12: R40.

    Article  Google Scholar 

  16. Curtis C, Shah SP, Chin SF, Turashvili G, Rueda OM, Dunning MJ et al. The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature 2012; 486: 346–352.

    Article  CAS  Google Scholar 

  17. Saal LH, Johansson P, Holm K, Gruvberger-Saal SK, She QB, Maurer M et al. Poor prognosis in carcinoma is associated with a gene expression signature of aberrant PTEN tumor suppressor pathway activity. Proc Natl Acad Sci USA 2007; 104: 7564–7569.

    Article  CAS  Google Scholar 

  18. Hanker AB, Pfefferle AD, Balko JM, Kuba MG, Young CD, Sanchez V et al. Mutant PIK3CA accelerates HER2-driven transgenic mammary tumors and induces resistance to combinations of anti-HER2 therapies. Proc Natl Acad Sci USA 2013; 110: 14372–14377.

    Article  CAS  Google Scholar 

  19. Yuan TL, Cantley LC . PI3K pathway alterations in cancer: variations on a theme. Oncogene 2008; 27: 5497–5510.

    Article  CAS  Google Scholar 

  20. Saini KS, Loi S, de Azambuja E, Metzger-Filho O, Saini ML, Ignatiadis M et al. Targeting the PI3K/AKT/mTOR and Raf/MEK/ERK pathways in the treatment of breast cancer. Cancer Treat Rev 2013; 39: 935–946.

    Article  CAS  Google Scholar 

  21. Wang ZP, Fu M, Wang LF, Liu JJ, Li YH, Brakebusch C et al. P21-activated kinase 1 (PAK1) can promote ERK. J Biol Chem 2013; 288: 20093–20099.

    Article  CAS  Google Scholar 

  22. King AJ, Sun HY, Diaz B, Barnard D, Miao WY, Bagrodia S et al. The protein kinase Pak3 positively regulates Raf-1 activity through phosphorylation of serine 338. Nature 1998; 396: 180–183.

    Article  CAS  Google Scholar 

  23. Slack-Davis JK, Eblen ST, Zecevic M, Boerner SA, Tarcsafalvi A, Diaz HB et al. PAK1 phosphorylation of MEK1 regulates fibronectin-stimulated MAPK activation. J Cell Biol 2003; 162: 281–291.

    Article  CAS  Google Scholar 

  24. Manser E, Leung T, Salihuddin H, Zhao ZS, Lim L . A brain serine threonine protein-kinase activated by Cdc42 and Rac1. Nature 1994; 367: 40–46.

    Article  CAS  Google Scholar 

  25. Fritsch R, de Krijger I, Fritsch K, George R, Reason B, Kumar MS et al. RAS and RHO families of GTPases directly regulate distinct phosphoinositide 3-kinase isoforms. Cell 2013; 153: 1050–1063.

    Article  CAS  Google Scholar 

  26. Yang HW, Shin MG, Lee S, Kim JR, Park WS, Cho KH et al. Cooperative activation of PI3K by Ras and Rho family small GTPases. Mol Cell 2012; 47: 281–290.

    Article  CAS  Google Scholar 

  27. Onesto C, Shutes A, Picard V, Schweighoffer F, Der CJ . Characterization of EHT 1864, a novel small molecule inhibitor of Rac family small GTPases. Small Gtpases Dis Part B 2008; 439: 111–129.

    Article  CAS  Google Scholar 

  28. Hill K, Krugmann S, Andrews SR, Coadwell WJ, Finan P, Welch HC et al. Regulation of P-Rex1 by phosphatidylinositol (3,4,5)-trisphosphate and Gbetagamma subunits. J Biol Chem 2005; 280: 4166–4173.

    Article  CAS  Google Scholar 

  29. Hoffman GR, Cerione RA . Signaling to the Rho GTPases: networking with the DH domain. FEBS Lett 2002; 513: 85–91.

    Article  CAS  Google Scholar 

  30. Hernandez-Negrete I, Carretero-Ortega J, Rosenfeldt H, Hernandez-Garcia R, Calderon-Salinas JV, Reyes-Cruz G et al. P-Rex1 links mammalian target of rapamycin signaling to Rac activation and cell migration. J Biol Chem 2007; 282: 23708–23715.

    Article  CAS  Google Scholar 

  31. Barretina J, Caponigro G, Stransky N, Venkatesan K, Margolin AA, Kim S et al. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 2012; 483: 603–607.

    Article  CAS  Google Scholar 

  32. O'Brien C, Wallin JJ, Sampath D, GuhaThakurta D, Savage H, Punnoose EA et al. Predictive biomarkers of sensitivity to the phosphatidylinositol 3' kinase inhibitor GDC-0941 in breast cancer preclinical models. Clin Cancer Res 2010; 16: 3670–3683.

    Article  CAS  Google Scholar 

  33. Ebi H, Costa C, Faber AC, Nishtala M, Kotani H, Juric D et al. PI3K regulates MEK/ERK signaling in breast cancer via the Rac-GEF, P-Rex1. Proc Natl Acad Sci USA 2013; 110: 21124–21129.

    Article  CAS  Google Scholar 

  34. Guillermet-Guibert J, Bjorklof K, Salpekar A, Gonella C, Ramadani F, Bilancio A et al. The p110 beta isoform of phosphoinositide 3-kinase signals downstream of G protein-coupled receptors and is functionally redundant with p110 gamma. Proc Natl Acad Sci USA 2008; 105: 8292–8297.

    Article  CAS  Google Scholar 

  35. Arias-Romero LE, Villamar-Cruz O, Pacheco A, Kosoff R, Huang M, Muthuswamy SK et al. A Rac-Pak signaling pathway is essential for ErbB2-mediated transformation of human breast epithelial cancer cells. Oncogene 2010; 29: 5839–5849.

    Article  CAS  Google Scholar 

  36. Premont RT, Perry SJ, Schmalzigaug R, Roseman JT, Xing Y, Claing A . The GIT/PIX complex: an oligomeric assembly of GIT family ARF GTPase-activating proteins and PIX family Rac1/Cdc42 guanine nucleotide exchange factors. Cell Signal 2004; 16: 1001–1011.

    Article  CAS  Google Scholar 

  37. Ong CC, Jubb AM, Haverty PM, Zhou W, Tran V, Truong T et al. Targeting p21-activated kinase 1 (PAK1) to induce apoptosis of tumor cells. Proc Natl Acad Sci USA 2011; 108: 7177–7182.

    Article  CAS  Google Scholar 

  38. Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature 2012; 490: 61–70.

    Article  Google Scholar 

  39. Cerami E, Gao J, Dogrusoz U, Gross BE, Sumer SO, Aksoy BA et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov 2012; 2: 401–404.

    Article  Google Scholar 

  40. Welch HC, Condliffe AM, Milne LJ, Ferguson GJ, Hill K, Webb LM et al. P-Rex1 regulates neutrophil function. Curr Biol 2005; 15: 1867–1873.

    Article  CAS  Google Scholar 

  41. Miller TW, Perez-Torres M, Narasanna A, Guix M, Stal O, Perez-Tenorio G et al. Loss of phosphatase and tensin homologue deleted on chromosome 10 engages ErbB3 and insulin-like growth factor-I receptor signaling to promote antiestrogen resistance in breast cancer. Cancer Res 2009; 69: 4192–4201.

    Article  CAS  Google Scholar 

  42. MacCoss MJ, McDonald WH, Saraf A, Sadygov R, Clark JM, Tasto JJ et al. Shotgun identification of protein modifications from protein complexes and lens tissue. Proc Natl Acad Sci USA 2002; 99: 7900–7905.

    Article  CAS  Google Scholar 

  43. Martinez MN, Emfinger CH, Overton M, Hill S, Ramaswamy TS, Cappel DA et al. Obesity and altered glucose metabolism impact HDL composition in CETP transgenic mice: a role for ovarian hormones. J Lipid Res 2012; 53: 379–389.

    Article  CAS  Google Scholar 

  44. Eng JK, McCormack AL, Yates JR III . An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J Am Soc Mass Spectrom 1994; 5: 976–989.

    Article  CAS  Google Scholar 

  45. Ma ZQ, Dasari S, Chambers MC, Litton MD, Sobecki SM, Zimmerman LJ et al. IDPicker 2.0: improved protein assembly with high discrimination peptide identification filtering. J Proteome Res 2009; 8: 3872–3881.

    Article  CAS  Google Scholar 

  46. Li J, Lu Y, Akbani R, Ju Z, Roebuck PL, Liu W et al. TCPA: a resource for cancer functional proteomics data. Nat Methods 2013; 10: 1046–1047.

    Article  CAS  Google Scholar 

  47. Hu ZY, Fan C, Oh DS, Marron JS, He XP, Qaqish BF et al. The molecular portraits of breast tumors are conserved across microarray platforms. BMC Genom 2006; 7: 96.

    Article  Google Scholar 

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Acknowledgements

We thank Chad Creighton for advice on gene expression analysis, and the Norris Cotton Cancer Center Transgenic and Genetic Construct Shared Resource and Genomics and Molecular Biology Shared Resource for assistance. This work was financially supported by NIH: K99/R00CA142899 (TWM), Dartmouth College Norris Cotton Cancer Center Support Grant P30CA023108, Vanderbilt-Ingram Cancer Center Breast Cancer Specialized Program of Research Excellence Grant P50CA98131 and Vanderbilt-Ingram Cancer Center Support Grant P30CA68485; the American Cancer Society IRG-58-009-50 (TWM) and 121329-RSG-11-187-01-TBG (to AMG); and The Commonwealth Foundation for Cancer Research (to AMG).

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Norris Cotton Cancer Center, Geisel School of Medicine at Dartmouth, Dartmouth-Hitchcock Medical Center, Lebanon, NH, USA

    L M Dillon, J R Bean, W Yang, K Shee, L K Symonds & T W Miller

  2. Department of Medicine, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA

    J M Balko & C L Arteaga

  3. Breast Cancer Research Program, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA

    J M Balko & C L Arteaga

  4. Proteomics Laboratory, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA

    W H McDonald

  5. Department of Breast Medical Oncology, MD Anderson Cancer Center, University of Texas, Houston, TX, USA

    S Liu & A M Gonzalez-Angulo

  6. Department of Systems Biology, MD Anderson Cancer Center, University of Texas, Houston, TX, USA

    S Liu, A M Gonzalez-Angulo & G B Mills

  7. Department of Cancer Biology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA

    C L Arteaga

  8. Comprehensive Breast Cancer Program, Norris Cotton Cancer Center, Geisel School of Medicine at Dartmouth, Lebanon, NH, USA

    T W Miller

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Dillon, L., Bean, J., Yang, W. et al. P-REX1 creates a positive feedback loop to activate growth factor receptor, PI3K/AKT and MEK/ERK signaling in breast cancer. Oncogene 34, 3968–3976 (2015). https://doi.org/10.1038/onc.2014.328

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