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FBW7 mutations mediate resistance of colorectal cancer to targeted therapies by blocking Mcl-1 degradation

Oncogene volume 36, pages 787–796 (2017)Cite this article

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Abstract

Colorectal cancer (CRC), the second leading cause of cancer-related deaths in the US, has been treated with targeted therapies. However, the mechanisms of differential responses and resistance of CRCs to targeted therapies are not well understood. In this study, we found that genetic alterations of FBW7, an E3 ubiquitin ligase and a tumor suppressor frequently mutated in CRCs, contribute to resistance to targeted therapies. CRC cells containing FBW7-inactivating mutations are insensitive to clinically used multi-kinase inhibitors of RAS/RAF/MEK/ERK signaling, including regorafenib and sorafenib. In contrast, sensitivity to these agents is not affected by oncogenic mutations in KRAS, BRAF, PIK3CA or p53. These cells are defective in apoptosis owing to blocked degradation of Mcl-1, a pro-survival Bcl-2 family protein. Deleting FBW7 in FBW7-wild-type CRC cells abolishes Mcl-1 degradation and recapitulates the in vitro and in vivo drug-resistance phenotypes of FBW7-mutant cells. CRC cells selected for regorafenib resistance have progressive enrichment of pre-existing FBW7 hotspot mutations, and are cross-resistant to other targeted drugs that induce Mcl-1 degradation. Furthermore, a selective Mcl-1 inhibitor restores regorafenib sensitivity in CRC cells with intrinsic or acquired resistance. Together, our results demonstrate FBW7 mutational status as a key genetic determinant of CRC response to targeted therapies, and Mcl-1 as an attractive therapeutic target.

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References

  1. Cunningham D, Atkin W, Lenz HJ, Lynch HT, Minsky B, Nordlinger B et al. Colorectal cancer. Lancet 2010; 375: 1030–1047.

    Article  PubMed  Google Scholar 

  2. Markowitz SD, Bertagnolli MM . Molecular origins of cancer: molecular basis of colorectal cancer. N Engl J Med 2009; 361: 2449–2460.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Vogelstein B, Kinzler KW . Cancer genes and the pathways they control. Nat Med 2004; 10: 789–799.

    CAS  PubMed  Google Scholar 

  4. Davis RJ, Welcker M, Clurman BE . Tumor suppression by the Fbw7 ubiquitin ligase: mechanisms and opportunities. Cancer Cell 2014; 26: 455–464.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Rajagopalan H, Jallepalli PV, Rago C, Velculescu VE, Kinzler KW, Vogelstein B et al. Inactivation of hCDC4 can cause chromosomal instability. Nature 2004; 428: 77–81.

    Article  CAS  PubMed  Google Scholar 

  6. Chu E . An update on the current and emerging targeted agents in metastatic colorectal cancer. Clin Colorectal Cancer 2012; 11: 1–13.

    Article  CAS  PubMed  Google Scholar 

  7. Demetri GD, Reichardt P, Kang YK, Blay JY, Rutkowski P, Gelderblom H et al. Efficacy and safety of regorafenib for advanced gastrointestinal stromal tumours after failure of imatinib and sunitinib (GRID): an international, multicentre, randomised, placebo-controlled, phase 3 trial. Lancet 2013; 381: 295–302.

    Article  CAS  PubMed  Google Scholar 

  8. Escudier B, Eisen T, Stadler WM, Szczylik C, Oudard S, Siebels M et al. Sorafenib in advanced clear-cell renal-cell carcinoma. N Engl J Med 2007; 356: 125–134.

    Article  CAS  PubMed  Google Scholar 

  9. Grothey A, Van Cutsem E, Sobrero A, Siena S, Falcone A, Ychou M et al. Regorafenib monotherapy for previously treated metastatic colorectal cancer (CORRECT): an international, multicentre, randomised, placebo-controlled, phase 3 trial. Lancet 2013; 381: 303–312.

    Article  CAS  PubMed  Google Scholar 

  10. Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 2008; 359: 378–390.

    Article  CAS  PubMed  Google Scholar 

  11. Wilhelm S, Carter C, Lynch M, Lowinger T, Dumas J, Smith RA et al. Discovery and development of sorafenib: a multikinase inhibitor for treating cancer. Nat Rev Drug Discov 2006; 5: 835–844.

    Article  CAS  PubMed  Google Scholar 

  12. Wilhelm SM, Dumas J, Adnane L, Lynch M, Carter CA, Schutz G et al. Regorafenib (BAY 73-4506): a new oral multikinase inhibitor of angiogenic, stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity. Int J Cancer 2011; 129: 245–255.

    Article  CAS  PubMed  Google Scholar 

  13. Bardelli A, Siena S . Molecular mechanisms of resistance to cetuximab and panitumumab in colorectal cancer. J Clin Oncol 2010; 28: 1254–1261.

    Article  CAS  PubMed  Google Scholar 

  14. Allegra CJ, Jessup JM, Somerfield MR, Hamilton SR, Hammond EH, Hayes DF et al. American Society of Clinical Oncology provisional clinical opinion: testing for KRAS gene mutations in patients with metastatic colorectal carcinoma to predict response to anti-epidermal growth factor receptor monoclonal antibody therapy. J Clin Oncol 2009; 27: 2091–2096.

    Article  PubMed  Google Scholar 

  15. Chan SL, Ma BB . An update on the safety and efficacy of regorafenib in the treatment of solid cancers. Exp Opin Drug Metab Toxicol 2014; 10: 1607–1614.

    Article  CAS  Google Scholar 

  16. Hata AN, Engelman JA, Faber AC . The BCL2 family: key mediators of the apoptotic response to targeted anticancer therapeutics. Cancer Discov 2015; 5: 475–487.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Bhola PD, Letai A . Mitochondria-judges and executioners of cell death sentences. Mol Cell 2016; 61: 695–704.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Thomas LW, Lam C, Edwards SW . Mcl-1; the molecular regulation of protein function. FEBS Lett 2010; 584: 2981–2989.

    Article  CAS  PubMed  Google Scholar 

  19. Mojsa B, Lassot I, Desagher S . Mcl-1 ubiquitination: unique regulation of an essential survival protein. Cells 2014; 3: 418–437.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Liu L, Cao Y, Chen C, Zhang X, McNabola A, Wilkie D et al. Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res 2006; 66: 11851–11858.

    Article  CAS  PubMed  Google Scholar 

  21. Rahmani M, Davis EM, Bauer C, Dent P, Grant S . Apoptosis induced by the kinase inhibitor BAY 43-9006 in human leukemia cells involves down-regulation of Mcl-1 through inhibition of translation. J Biol Chem 2005; 280: 35217–35227.

    Article  CAS  PubMed  Google Scholar 

  22. Wertz IE, Kusam S, Lam C, Okamoto T, Sandoval W, Anderson DJ et al. Sensitivity to antitubulin chemotherapeutics is regulated by MCL1 and FBW7. Nature 2011; 471: 110–114.

    Article  CAS  PubMed  Google Scholar 

  23. Yu C, Bruzek LM, Meng XW, Gores GJ, Carter CA, Kaufmann SH et al. The role of Mcl-1 downregulation in the proapoptotic activity of the multikinase inhibitor BAY 43-9006. Oncogene 2005; 24: 6861–6869.

    Article  CAS  PubMed  Google Scholar 

  24. Zhong Q, Gao W, Du F, Wang X . Mule/ARF-BP1, a BH3-only E3 ubiquitin ligase, catalyzes the polyubiquitination of Mcl-1 and regulates apoptosis. Cell 2005; 121: 1085–1095.

    Article  CAS  PubMed  Google Scholar 

  25. Maurer U, Charvet C, Wagman AS, Dejardin E, Green DR . Glycogen synthase kinase-3 regulates mitochondrial outer membrane permeabilization and apoptosis by destabilization of MCL-1. Mol Cell 2006; 21: 749–760.

    Article  CAS  PubMed  Google Scholar 

  26. Inuzuka H, Shaik S, Onoyama I, Gao D, Tseng A, Maser RS et al. SCF(FBW7) regulates cellular apoptosis by targeting MCL1 for ubiquitylation and destruction. Nature 2011; 471: 104–109.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Chen D, Wei L, Yu J, Zhang L . Regorafenib inhibits colorectal tumor growth through PUMA-mediated apoptosis. Clin Cancer Res 2014; 20: 3472–3484.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Varadarajan S, Vogler M, Butterworth M, Dinsdale D, Walensky LD, Cohen GM . Evaluation and critical assessment of putative MCL-1 inhibitors. Cell Death Differ 2013; 20: 1475–1484.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Tahir SK, Yang X, Anderson MG, Morgan-Lappe SE, Sarthy AV, Chen J et al. Influence of Bcl-2 family members on the cellular response of small-cell lung cancer cell lines to ABT-737. Cancer Res 2007; 67: 1176–1183.

    Article  CAS  PubMed  Google Scholar 

  30. Min SH, Lau AW, Lee TH, Inuzuka H, Wei S, Huang P et al. Negative regulation of the stability and tumor suppressor function of Fbw7 by the Pin1 prolyl isomerase. Mol Cell 2012; 46: 771–783.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. O'Neil J, Grim J, Strack P, Rao S, Tibbitts D, Winter C et al. FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to gamma-secretase inhibitors. J Exp Med 2007; 204: 1813–1824.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Thompson BJ, Buonamici S, Sulis ML, Palomero T, Vilimas T, Basso G et al. The SCFFBW7 ubiquitin ligase complex as a tumor suppressor in T cell leukemia. J Exp Med 2007; 204: 1825–1835.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. He L, Torres-Lockhart K, Forster N, Ramakrishnan S, Greninger P, Garnett MJ et al. Mcl-1 and FBW7 control a dominant survival pathway underlying HDAC and Bcl-2 inhibitor synergy in squamous cell carcinoma. Cancer Discov 2013; 3: 324–337.

    Article  CAS  PubMed  Google Scholar 

  34. Deshaies RJ, Joazeiro CA . RING domain E3 ubiquitin ligases. Ann Rev Biochem 2009; 78: 399–434.

    Article  CAS  PubMed  Google Scholar 

  35. Bhattacharyya S, Yu H, Mim C, Matouschek A . Regulated protein turnover: snapshots of the proteasome in action. Nat Rev Mol Cell Biol 2014; 15: 122–133.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Balamurugan K, Wang JM, Tsai HH, Sharan S, Anver M, Leighty R et al. The tumour suppressor C/EBPdelta inhibits FBXW7 expression and promotes mammary tumour metastasis. EMBO J 2010; 29: 4106–4117.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Lerner M, Lundgren J, Akhoondi S, Jahn A, Ng HF, Akbari Moqadam F et al. MiRNA-27a controls FBW7/hCDC4-dependent cyclin E degradation and cell cycle progression. Cell Cycle 2011; 10: 2172–2183.

    Article  CAS  PubMed  Google Scholar 

  38. Schulein-Volk C, Wolf E, Zhu J, Xu W, Taranets L, Hellmann A et al2014. Dual regulation of Fbw7 function and oncogenic transformation by Usp28. Cell Rep 9: 1099–1109.

    Article  PubMed  Google Scholar 

  39. Ding Q, He X, Hsu JM, Xia W, Chen CT, Li LY et al. Degradation of Mcl-1 by beta-TrCP mediates glycogen synthase kinase 3-induced tumor suppression and chemosensitization. Mol Cell Biol 2007; 27: 4006–4017.

    Article  CAS  PubMed  Google Scholar 

  40. Schwickart M, Huang X, Lill JR, Liu J, Ferrando R, French DM et al. Deubiquitinase USP9X stabilizes MCL1 and promotes tumour cell survival. Nature 2010; 463: 103–107.

    Article  CAS  PubMed  Google Scholar 

  41. Choudhary GS, Tat TT, Misra S, Hill BT, Smith MR, Almasan A et al. Cyclin E/Cdk2-dependent phosphorylation of Mcl-1 determines its stability and cellular sensitivity to BH3 mimetics. Oncotarget 2015; 6: 16912–16925.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Diaz LA Jr, Williams RT, Wu J, Kinde I, Hecht JR, Berlin J et al. The molecular evolution of acquired resistance to targeted EGFR blockade in colorectal cancers. Nature 2012; 486: 537–540.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Misale S, Yaeger R, Hobor S, Scala E, Janakiraman M, Liska D et al. Emergence of KRAS mutations and acquired resistance to anti-EGFR therapy in colorectal cancer. Nature 2012; 486: 532–536.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Peng R, Tong JS, Li H, Yue B, Zou F, Yu J et al. Targeting Bax interaction sites reveals that only homo-oligomerization sites are essential for its activation. Cell Death Differ 2013; 20: 744–754.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Yu J, Wang Z, Kinzler KW, Vogelstein B, Zhang L . PUMA mediates the apoptotic response to p53 in colorectal cancer cells. Proc Natl Acad Sci USA 2003; 100: 1931–1936.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Dudgeon C, Peng R, Wang P, Sebastiani A, Yu J, Zhang L . Inhibiting oncogenic signaling by sorafenib activates PUMA via GSK3beta and NF-kappaB to suppress tumor cell growth. Oncogene 2012; 31: 4848–4858.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Chen D, Ming L, Zou F, Peng Y, Van Houten B, Yu J et al. TAp73 promotes cell survival upon genotoxic stress by inhibiting p53 activity. Oncotarget 2014; 5: 8107–8122.

    PubMed  PubMed Central  Google Scholar 

  48. Qiu W, Wang X, Leibowitz B, Liu H, Barker N, Okada H et al. Chemoprevention by nonsteroidal anti-inflammatory drugs eliminates oncogenic intestinal stem cells via SMAC-dependent apoptosis. Proc Natl Acad Sci USA 2010; 107: 20027–20032.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank our lab members for critical reading. This work is supported by U.S. National Institute of Health grants (CA106348, CA172136 and CA203028 to LZ; U01DK085570 and AI068021to JY) and American Cancer Society grant (RGS-10-124-01-CCE to JY). This project used the UPCI shared facilities that were supported in part by award P30CA047904.

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Authors and Affiliations

  1. Department of Pharmacology and Chemical Biology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    J Tong, S Tan & L Zhang

  2. College of Life Sciences, Sichuan University, Chengdu, China

    S Tan & F Zou

  3. Department of Pathology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    J Yu

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Correspondence to L Zhang.

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Tong, J., Tan, S., Zou, F. et al. FBW7 mutations mediate resistance of colorectal cancer to targeted therapies by blocking Mcl-1 degradation. Oncogene 36, 787–796 (2017). https://doi.org/10.1038/onc.2016.247

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