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A novel proteotoxic combination therapy for EGFR+ and HER2+ cancers

Oncogene volume 38, pages 4264–4282 (2019)Cite this article

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Abstract

While HER2 and EGFR are overexpressed in breast cancers and multiple other types of tumors, the use of EGFR and/or HER2 inhibitors have failed to cure many cancer patients, largely because cancers acquire resistance to HER2/EGFR-specific drugs. Cancers that overexpress the HER-family proteins EGFR, HER2, and HER3 are uniquely sensitive to agents that disrupt HER2 and EGFR protein folding. We previously showed that disruption of disulfide bond formation by Disulfide Disrupting Agents (DDAs) kills HER2/EGFR overexpressing cells through multiple mechanisms. Herein, we show that interference with proline isomerization in HER2/EGFR overexpressing cells also induces cancer cell death. The peptidyl-prolyl isomerase inhibitor Cyclosporine A (CsA) selectively kills EGFR+ or HER2+ breast cancer cells in vitro by activating caspase-dependent apoptotic pathways. Further, CsA synergizes with the DDA tcyDTDO to kill HER2/EGFR overexpressing cells in vitro and the two agents cooperate to kill HER2+ tumors in vivo. There is a critical need for novel strategies to target HER2+ and EGFR+ cancers that are resistant to currently available mechanism-based agents. Drugs that target HER2/EGFR protein folding, including DDAs and CsA, have the potential to kill cancers that overexpress EGFR or HER2 through the induction of proteostatic synthetic lethality.

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References

  1. Chlebowski RT, Manson JE, Anderson GL, Cauley JA, Aragaki AK, Stefanick ML, et al. Estrogen plus progestin and breast cancer incidence and mortality in the Women’s Health Initiative Observational Study. J Natl Cancer Inst. 2013;105:526–35.

    Article  CAS  Google Scholar 

  2. Obeng EA, Carlson LM, Gutman DM, Harrington WJ Jr., Lee KP, Boise LH. Proteasome inhibitors induce a terminal unfolded protein response in multiple myeloma cells. Blood. 2006;107:4907–16.

    Article  CAS  Google Scholar 

  3. Wang M, Law ME, Castellano RK, Law BK. The unfolded protein response as a target for anticancer therapeutics. Crit Rev Oncol/Hematol. 2018;127:66–79.

    Article  Google Scholar 

  4. Stocki P, Chapman DC, Beach LA, Williams DB. Depletion of cyclophilins B and C leads to dysregulation of endoplasmic reticulum redox homeostasis. J Biol Chem. 2014;289:23086–96.

    Article  CAS  Google Scholar 

  5. Dunyak BM, Gestwicki JE. Peptidyl-proline isomerases (PPIases): targets for natural products and natural product-inspired compounds. J Med Chem. 2016;59:9622–44.

    Article  CAS  Google Scholar 

  6. Theuerkorn M, Fischer G, Schiene-Fischer C. Prolyl cis/trans isomerase signalling pathways in cancer. Curr Opin Pharmacol. 2011;11:281–7.

    Article  CAS  Google Scholar 

  7. Werneck MB, Hottz E, Bozza PT, Viola JP. Cyclosporin A inhibits colon cancer cell growth independently of the calcineurin pathway. Cell Cycle. 2012;11:3997–4008.

    Article  CAS  Google Scholar 

  8. Ciechomska IA, Gabrusiewicz K, Szczepankiewicz AA, Kaminska B. Endoplasmic reticulum stress triggers autophagy in malignant glioma cells undergoing cyclosporine a-induced cell death. Oncogene. 2013;32:1518–29.

    Article  CAS  Google Scholar 

  9. Robinson PJ, Pringle MA, Woolhead CA, Bulleid NJ. Folding of a single domain protein entering the endoplasmic reticulum precedes disulfide formation. J Biol Chem. 2017;292:6978–86.

    Article  CAS  Google Scholar 

  10. Lemmon MA, Schlessinger J. Cell signaling by receptor tyrosine kinases. Cell. 2010;141:1117–34.

    Article  CAS  Google Scholar 

  11. Ogiso H, Ishitani R, Nureki O, Fukai S, Yamanaka M, Kim JH, et al. Crystal structure of the complex of human epidermal growth factor and receptor extracellular domains. Cell. 2002;110:775–87.

    Article  CAS  Google Scholar 

  12. Garrett TP, McKern NM, Lou M, Elleman TC, Adams TE, Lovrecz GO, et al. The crystal structure of a truncated ErbB2 ectodomain reveals an active conformation, poised to interact with other ErbB receptors. Mol Cell. 2003;11:495–505.

    Article  CAS  Google Scholar 

  13. Cho HS, Mason K, Ramyar KX, Stanley AM, Gabelli SB, Denney DW Jr., et al. Structure of the extracellular region of HER2 alone and in complex with the Herceptin Fab. Nature. 2003;421:756–60.

    Article  CAS  Google Scholar 

  14. Cho HS, Leahy DJ. Structure of the extracellular region of HER3 reveals an interdomain tether. Science. 2002;297:1330–3.

    Article  CAS  Google Scholar 

  15. Ferreira RB, Law ME, Jahn SC, Davis BJ, Heldermon CD, Reinhard M, et al. Novel agents that downregulate EGFR, HER2, and HER3 in parallel. Oncotarget. 2015;6:10445–59.

    Article  Google Scholar 

  16. Ferreira RB, Wang M, Law ME, Davis BJ, Bartley AN, Higgins PJ, et al. Disulfide bond disrupting agents activate the unfolded protein response in EGFR- and HER2-positive breast tumor cells. Oncotarget. 2017;8:28971–89.

    Article  Google Scholar 

  17. Gyorffy B, Lanczky A, Eklund AC, Denkert C, Budczies J, Li Q, et al. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. Breast Cancer Res Treat. 2010;123:725–31.

    Article  Google Scholar 

  18. Iyer RP, Egan W, Regan JB, Beaucage SL. 3h-1,2-Benzodithiole-3-One 1,1-Dioxide as an improved sulfurizing reagent in the solid-phase synthesis of oligodeoxyribonucleoside phosphorothioates. J Am Chem Soc. 1990;112:1253–4.

    Article  CAS  Google Scholar 

  19. Srivastava PK, Field L. Organic disulfides and related substances .36. some oxodisulfide cleavage reactions to form disulfides and trisulfides. J Org Chem. 1972;37:4196–8.

    Article  CAS  Google Scholar 

  20. Bass SW, Evans SA. C-13 Nuclear magnetic-resonance spectral properties of alkyl disulfides, thiolsulfinates, and thiolsulfonates. J Org Chem. 1980;45:710–5.

    Article  CAS  Google Scholar 

  21. Goethals EJ, Huylebro J, Smolders W. Synthesis of thiosultones. B Soc Chim Belg. 1969;78:191–&.

    Article  CAS  Google Scholar 

  22. Hartman DA. Determination of the stability of drugs in plasma. Curr Protoc Pharmacol. 2003;Unit 7:6. Chapter 7

    PubMed  Google Scholar 

  23. Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70:440–6.

    Article  CAS  Google Scholar 

  24. Shimizu T, Martin MS, Pelletier H, Lagadec P, Martin F. Effects of cyclosporin A on progressive and regressive tumors induced by two cancer lines derived from a single colon carcinoma chemically induced in the rat. Immunobiology. 1989;178:401–15.

    Article  CAS  Google Scholar 

  25. DeRose YS, Gligorich KM, Wang G, Georgelas A, Bowman P, Courdy SJ, et al. Patient-derived models of human breast cancer: protocols for in vitro and in vivo applications in tumor biology and translational medicine. Curr Protoc Pharmacol. 2013;Unit14:23. Chapter 14

    Google Scholar 

  26. Neckers L. Effects of geldanamycin and other naturally occurring small molecule antagonists of heat shock protein 90 on HER2 protein expression. Breast Dis. 2000;11:49–59.

    Article  CAS  Google Scholar 

  27. Takenokuchi M, Miyamoto K, Saigo K, Taniguchi T, Bortezomib Causes ER. Stress-related death of acute promyelocytic leukemia cells through excessive accumulation of PML-RARA. Anticancer Res. 2015;35:3307–16.

    CAS  PubMed  Google Scholar 

  28. Anderson DJ, Le Moigne R, Djakovic S, Kumar B, Rice J, Wong S, et al. Targeting the AAA ATPase p97 as an approach to treat cancer through disruption of protein homeostasis. Cancer Cell. 2015;28:653–65.

    Article  CAS  Google Scholar 

  29. Vekaria PH, Home T, Weir S, Schoenen FJ, Rao R. Targeting p97 to disrupt protein homeostasis in cancer. Front Oncol. 2016;6:181.

    Article  Google Scholar 

  30. Kallioniemi OP, Kallioniemi A, Kurisu W, Thor A, Chen LC, Smith HS, et al. ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization. Proc Natl Acad Sci USA. 1992;89:5321–5.

    Article  CAS  Google Scholar 

  31. Field L, Khim YH. Organic disulfides and related substances. 33. sodium 4-(2-acetamidoethyldithio)butanesulfinate and related compounds as antiradiation drugs. J Med Chem. 1972;15:312–5.

    Article  CAS  Google Scholar 

  32. Hysing J, Wist E. Cardiotoxic effects of trastuzumab. Tidsskr Nor Laege. 2011;131:2239–41.

    Article  Google Scholar 

  33. Kopper L. Lapatinib: a sword with two edges. Pathol Oncol Res. 2008;14:1–8.

    Article  CAS  Google Scholar 

  34. Erdogan E, Lamark T, Stallings-Mann M, Lee J, Pellecchia M, Thompson EA, et al. Aurothiomalate inhibits transformed growth by targeting the PB1 domain of protein kinase Ciota. J Biol Chem. 2006;281:28450–9.

    Article  CAS  Google Scholar 

  35. Feng X, Holmlund T, Zheng C, Fadeel B. Proapoptotic effects of the novel proteasome inhibitor b-AP15 on multiple myeloma cells and natural killer cells. Exp Hematol. 2014;42:172–82.

    Article  CAS  Google Scholar 

  36. Singh N, Joshi R, Komurov K. HER2-mTOR signaling-driven breast cancer cells require ER-associated degradation to survive. Sci Signal. 2015;8:ra52.

    Article  Google Scholar 

  37. Ni S, Yuan Y, Huang J, Mao X, Lv M, Zhu J, et al. Discovering potent small molecule inhibitors of cyclophilin A using de novo drug design approach. J Med Chem. 2009;52:5295–8.

    Article  CAS  Google Scholar 

  38. Ahmed-Belkacem A, Colliandre L, Ahnou N, Nevers Q, Gelin M, Bessin Y, et al. Fragment-based discovery of a new family of non-peptidic small-molecule cyclophilin inhibitors with potent antiviral activities. Nat Commun. 2016;7:12777.

    Article  CAS  Google Scholar 

  39. Rhodes DR, Kalyana-Sundaram S, Mahavisno V, Varambally R, Yu J, Briggs BB, et al. Oncomine 3.0: genes, pathways, and networks in a collection of 18,000 cancer gene expression profiles. Neoplasia. 2007;9:166–80.

    Article  CAS  Google Scholar 

  40. Garg AD, Agostinis P. ER stress, autophagy and immunogenic cell death in photodynamic therapy-induced anti-cancer immune responses. Photochem Photobiol Sci. 2014;13:474–87.

    Article  CAS  Google Scholar 

  41. Law ME, Ferreira RB, Davis BJ, Higgins PJ, Kim JS, Castellano RK, et al. CUB domain-containing protein 1 and the epidermal growth factor receptor cooperate to induce cell detachment. Breast Cancer Res. 2016;18:80.

    Article  Google Scholar 

  42. Liu X, Ory V, Chapman S, Yuan H, Albanese C, Kallakury B, et al. ROCK inhibitor and feeder cells induce the conditional reprogramming of epithelial cells. Am J Pathol. 2012;180:599–607.

    Article  CAS  Google Scholar 

  43. Law BK, Waltner-Law ME, Entingh AJ, Chytil A, Aakre ME, Norgaard P, et al. Salicylate-induced growth arrest is associated with inhibition of p70s6k and down-regulation of c-myc, cyclin D1, cyclin A, and proliferating cell nuclear antigen. J Biol Chem. 2000;275:38261–7.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank breast cancer research advocate Mrs. Jeri Francoeur for her advice and support.

Funding

This work was supported by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program under Award Nos. W81XWH-15-1-0199 (BL) and W81XWH-15-1-0200 (RC). These studies were supported in part by grants from the Florida Breast Cancer Foundation (BL and RC), The Ocala Royal Dames for Cancer Research (BL), the Florida Department of Health Bankhead-Coley Program (4BF03, BL), the Collaboration of Scientists for Critical Research in Biomedicine (CSCRB, Inc., BL), and a UF-Health Cancer Center pilot project award (BL, RC, and CH). RF is grateful to the University of Florida for a Graduate School Fellowship.

Author contributions

MW, RF, ML, EY, AG, AS, BA, RC, and BL planned the studies. MW, RF, ML, BD, EY, AG, AS, and BL carried out the studies. BA, ER, CC, CH, and SN provided advice and reagents necessary for carrying out the studies. MW, RF, ML, EY, AG, AS, BA, ER, CC, SN, CH, RC, and BL edited the manuscript.

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Authors and Affiliations

  1. Department of Pharmacology, University of Florida, Gainesville, FL, 32610, USA

    Mengxiong Wang, Mary E. Law, Bradley J. Davis, Edgardo Rodriguez & Brian K. Law

  2. Department of Chemistry, University of Florida, Gainesville, FL, 32611, USA

    Renan B. Ferreira, Elham Yaaghubi, Amanda F. Ghilardi & Ronald K. Castellano

  3. Department of Pharmaceutics, University of Florida, Gainesville, FL, 32610, USA

    Abhisheak Sharma & Bonnie A. Avery

  4. Institute of Molecular Medicine, College of Medicine and Center for Infectious Disease and Signaling Research, National Cheng Kung University, Tainan, Taiwan

    Chi-Wu Chiang

  5. Department of Anatomy and Cell Biology, University of Florida, Gainesville, FL, 32610, USA

    Satya Narayan

  6. UF-Health Cancer Center, University of Florida, Gainesville, FL, 32610, USA

    Satya Narayan, Coy D. Heldermon, Ronald K. Castellano & Brian K. Law

  7. Department of Medicine, University of Florida, Gainesville, FL, 32610, USA

    Coy D. Heldermon

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Correspondence to Ronald K. Castellano or Brian K. Law.

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Wang, M., Ferreira, R.B., Law, M.E. et al. A novel proteotoxic combination therapy for EGFR+ and HER2+ cancers. Oncogene 38, 4264–4282 (2019). https://doi.org/10.1038/s41388-019-0717-6

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