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Cancer Chemotherapy and Pharmacology

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01-02-2017 | Original Article

Reduced proliferation and colony formation of head and neck squamous cell carcinoma (HNSCC) after dual targeting of EGFR and hedgehog pathways

Authors: Hannes Liebig, Georg Günther, Marlen Kolb, Christian Mozet, Andreas Boehm, Andreas Dietz, Gunnar Wichmann

Published in: Cancer Chemotherapy and Pharmacology | Issue 2/2017

Abstract

Purpose

The hedgehog signalling pathway (Hh) is frequently active in head and neck squamous cell carcinoma (HNSCC). Overexpressed Hh associates with poor prognosis. The Hh inhibitor vismodegib targets smoothened, and based on molecular data, may prevent resistance to EGFR targeting.

Methods

To elucidate potential roles of vismodegib in HNSCC therapy, its sole effects and those combined with cisplatin, docetaxel, and cetuximab on HNSCC cell lines were assessed by MTT metabolisation and BrdU incorporation. Colony formation (CF) of primary HNSCC cells was studied utilizing the FLAVINO-protocol. Combinatory effects were analysed regarding antagonism, additivity or synergism. Associations between the ex vivo detected mode of action of vismodegib with other treatments related to patient characteristics were assessed and progression-free survival (PFS) in patient groups compared using Kaplan–Meier curves.

Results

Vismodegib suppressed BrdU incorporation significantly stronger than MTT turnover; CF was significantly inhibited at ≥20 µM vismodegib while concentrations <20 µM acted hormetic. Combining 20 µM vismodegib plus docetaxel (T), cisplatin (P), and cetuximab (E), additively enhanced anti-tumoral activity in HNSCC samples from patients with superior PFS highlighting a potential role for ex-vivo testing of this combination for use as a prognostic classifier.

Conclusion

We provide ex-vivo evidence for vismodegib’s potential in HNSCC therapies, especially if combined with cetuximab, cisplatin and docetaxel.

Zigon G, Berrino F, Gatta G et al (2011) Prognoses for head and neck cancers in Europe diagnosed in 1995–1999: a population-based study. Ann Oncol 22:165–174. doi:10.1093/annonc/mdq306 CrossRefPubMed

Ferlay J, Steliarova-Foucher E, Lortet-Tieulent J et al (2013) Cancer incidence and mortality patterns in Europe: estimates for 40 countries in 2012. Eur J Cancer 49:1374–1403. doi:10.1016/j.ejca.2012.12.027 CrossRefPubMed

Macfarlane GJ, Zheng T, Marshall JR et al (1995) Alcohol, tobacco, diet and the risk of oral cancer. A pooled analysis of three case–control studies. Eur J Cancer Part B Oral Oncol 31:181–187. doi:10.1016/0964-1955(95)00005-3 CrossRef

Andl T, Kahn T, Pfuhl A et al (1998) Etiological involvement of oncogenic human papillomavirus in tonsillar squamous cell carcinomas lacking retinoblastoma cell cycle control. Cancer Res 58:5–13PubMed

Gillison ML (2000) Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Ntnl Cancer Inst 92:709–720. doi:10.1093/jnci/92.9.709 CrossRef

Seshacharyulu P, Ponnusamy MP, Haridas D, Jain M, Ganti AK, Batra SK (2012) Targeting the EGFR signaling pathway in cancer therapy. Expert Opin Ther Targets 16:15–31. doi:10.1517/14728222.2011.648617 CrossRefPubMedPubMedCentral

Grandis JR, Tweardy DJ (1993) Elevated levels of transforming growth factor alpha and epidermal growth factor receptor messenger RNA are early markers of carcinogenesis in head and neck cancer. Cancer Res 53:3579–3584PubMed

Bonner JA, Harari PM, Giralt J et al (2006) Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. N Engl J Med 354:567–578. doi:10.1056/NEJMoa053422 CrossRefPubMed

Rivera F, García-Castaño A, Vega N, Vega-Villegas ME, Gutiérrez-Sanz L (2009) Cetuximab in metastatic or recurrent head and neck cancer: the EXTREME trial. Expert Rev Anticancer Ther 9:1421–1428. doi:10.1586/era.09.113 CrossRefPubMed

10.

Dietz A, Boehm A, Mozet C, Wichmann G, Giannis A (2008) Current aspects of targeted therapy in head and neck tumors. Eur Arch Otorhinolaryngol 265(Suppl 1):S3–S12. doi:10.1007/s00405-008-0697-6 CrossRefPubMed

11.

Scales SJ, Sauvage FJ de (2009) Mechanisms of hedgehog pathway activation in cancer and implications for therapy. Trends Pharmacol Sci 30:303–312. doi:10.1016/j.tips.2009.03.007 CrossRefPubMed

12.

Fujii K, Miyashita T (2014) Gorlin syndrome (nevoid basal cell carcinoma syndrome): update and literature review. Pediatr Int 56:667–674. doi:10.1111/ped.12461 CrossRefPubMed

13.

Romer J, Curran T (2005) Targeting medulloblastoma: small-molecule inhibitors of the sonic hedgehog pathway as potential cancer therapeutics. Cancer Res 65:4975–4978. doi:10.1158/0008-5472.CAN-05-0481 CrossRefPubMed

14.

Varnat F, Duquet A, Malerba M et al (2009) Human colon cancer epithelial cells harbour active HEDGEHOG-GLI signalling that is essential for tumour growth, recurrence, metastasis and stem cell survival and expansion. EMBO Mol Med 1:338–351. doi:10.1002/emmm.200900039 CrossRefPubMedPubMedCentral

15.

Berman DM, Karhadkar SS, Maitra A et al (2003) Widespread requirement for hedgehog ligand stimulation in growth of digestive tract tumours. Nature 425:846–851. doi:10.1038/nature01972 CrossRefPubMed

16.

Watkins DN, Berman DM, Burkholder SG, Wang B, Beachy PA, Baylin SB (2003) Hedgehog signalling within airway epithelial progenitors and in small-cell lung cancer. Nature 422:313–317. doi:10.1038/nature01493 CrossRefPubMed

17.

Karhadkar SS, Bova GS, Abdallah N et al (2004) Hedgehog signalling in prostate regeneration, neoplasia and metastasis. Nature 431:707–712. doi:10.1038/nature02962 CrossRefPubMed

18.

Chung CH, Dignam JJ, Hammond ME et al (2011) Glioma-associated oncogene family zinc finger 1 expression and metastasis in patients with head and neck squamous cell carcinoma treated with radiation therapy (RTOG 9003). J Clin Oncol 29:1326–1334. doi:10.1200/JCO.2010.32.3295 CrossRefPubMedPubMedCentral

19.

Dimitrova K, Stoehr M, Dehghani F et al (2013) Overexpression of the hedgehog signalling pathway in head and neck squamous cell carcinoma. Onkologie 36:279–286. doi:10.1159/000350322 CrossRefPubMed

20.

Fertig EJ, Markovic A, Danilova LV et al (2013) Preferential activation of the hedgehog pathway by epigenetic modulations in HPV negative HNSCC identified with meta-pathway analysis. PLoS One 8:e78127. doi:10.1371/journal.pone.0078127 CrossRefPubMedPubMedCentral

21.

Keysar SB, Le PN, Anderson RT et al (2013) Hedgehog signaling alters reliance on EGF receptor signaling and mediates anti-EGFR therapeutic resistance in head and neck cancer. Cancer Res 73:3381–3392. doi:10.1158/0008-5472.CAN-12-4047 CrossRefPubMedPubMedCentral

22.

Gan GN, Eagles J, Keysar SB et al (2014) Hedgehog signaling drives radioresistance and stroma-driven tumor repopulation in head and neck squamous cancers. Cancer Res 74:7024–7036. doi:10.1158/0008-5472.CAN-14-1346 CrossRefPubMedPubMedCentral

23.

Mozet C, Stoehr M, Dimitrova K, Dietz A, Wichmann G (2013) Hedgehog targeting by cyclopamine suppresses head and neck squamous cell carcinoma and enhances chemotherapeutic effects. Anticancer Res 33:2415–2424PubMed

24.

Sekulic A, Migden MR, Oro AE et al (2012) Efficacy and safety of vismodegib in advanced basal-cell carcinoma. N Engl J Med 366:2171–2179. doi:10.1056/NEJMoa1113713 CrossRefPubMedPubMedCentral

25.

Dietz A, Tschöp K, Wichmann G, Granzow C (2015) Method and kit for the ex vivo evaluation of the response of a tumor to conditions to be tested 09.04.2009. Accessed 10 Sep 2015

26.

Dietz A, Boehm A, Horn I et al (2010) Assay-based response evaluation in head and neck oncology: requirements for better decision making. Eur Arch Otorhinolaryngol 267:483–494. doi:10.1007/s00405-009-1191-5 CrossRefPubMed

27.

Horn I, Wichmann G, Mozet C et al (2010) Heterogeneity of epithelial and stromal cells of head and neck squamous cell carcinomas in ex vivo chemoresponse. Cancer Chemother Pharmacol 65:1153–1163. doi:10.1007/s00280-009-1124-4 CrossRefPubMed

28.

Stoehr M, Mozet C, Boehm A, Aigner A, Dietz A, Wichmann G (2014) Simvastatin suppresses head and neck squamous cell carcinoma ex vivo and enhances the cytostatic effects of chemotherapeutics. Cancer Chemother Pharmacol 73:827–837. doi:10.1007/s00280-014-2412-1 CrossRefPubMed

29.

Schrader C, Boehm A, Reiche A, Dietz A, Mozet C, Wichmann G (2012) Combined effects of lapatinib and cisplatin on colony formation of head and neck squamous cell carcinoma. Anticancer Res 32:3191–3199PubMed

30.

Zhao M, Sano D, Pickering CR et al (2011) Assembly and initial characterization of a panel of 85 genomically validated cell lines from diverse head and neck tumor sites. Clin Cancer Res 17:7248–7264. doi:10.1158/1078-0432.CCR-11-0690 CrossRefPubMedPubMedCentral

31.

Sobin LH, Gospodarowicz MK, Wittekind C (2010) TNM classification of malignant tumours. Wiley, Oxford

32.

Qualtrough D, Rees P, Speight B, Williams AC, Paraskeva C (2015) The hedgehog inhibitor cyclopamine reduces β-catenin-Tcf transcriptional activity, induces E-cadherin expression, and reduces invasion in colorectal cancer cells. Cancers 7:1885–1899. doi:10.3390/cancers7030867 CrossRefPubMedPubMedCentral

33.

Yoo YA, Kang MH, Lee HJ et al (2011) Sonic hedgehog pathway promotes metastasis and lymphangiogenesis via activation of Akt, EMT, and MMP-9 pathway in gastric cancer. Cancer Res 71:7061–7070. doi:10.1158/0008-5472.CAN-11-1338 CrossRefPubMed

34.

Bora-Singhal N, Perumal D, Nguyen J, Chellappan S (2015) Gli1-mediated regulation of Sox2 facilitates self-renewal of stem-like cells and confers resistance to EGFR inhibitors in non-small cell lung cancer. Neoplasia (New York, N.Y.) 17:538–551. doi:10.1016/j.neo.2015.07.001 CrossRef

35.

Zahreddine HA, Culjkovic-Kraljacic B, Assouline S et al (2014) The sonic hedgehog factor GLI1 imparts drug resistance through inducible glucuronidation. Nature 511:90–93. doi:10.1038/nature13283 CrossRefPubMedPubMedCentral

36.

Della Corte CM, Bellevicine C, Vicidomini G et al (2015) SMO gene amplification and activation of the hedgehog pathway as novel mechanisms of resistance to anti-epidermal growth factor receptor drugs in human lung cancer. Clin Cancer Res 21:4686–4697. doi:10.1158/1078-0432.CCR-14-3319 CrossRefPubMed

37.

Bowles DW, Keysar SB, Glogowska MJ et al (2014) A pilot study of cetuximab and the hedgehog inhibitor IPI-926 in recurrent/metastatic (R/M) head-and-neck squamous cell carcinoma. Int J Radiat Oncol Biol Phys 88:511. doi:10.1016/j.ijrobp.2013.11.152 CrossRef

38.

Singh S, Wang Z, Liang Fei D et al (2011) Hedgehog-producing cancer cells respond to and require autocrine hedgehog activity. Cancer Res 71:4454–4463. doi:10.1158/0008-5472.CAN-10-2313 CrossRefPubMed

39.

Yauch RL, Gould SE, Scales SJ et al (2008) A paracrine requirement for hedgehog signalling in cancer. Nature 455:406–410. doi:10.1038/nature07275 CrossRefPubMed

40.

Rhim AD, Oberstein PE, Thomas DH et al (2014) Stromal elements act to restrain, rather than support, pancreatic ductal adenocarcinoma. Cancer cell 25:735–747. doi:10.1016/j.ccr.2014.04.021 CrossRefPubMedPubMedCentral

41.

Chun SG, Park H, Pandita RK et al (2015) Targeted inhibition of histone deacetylases and hedgehog signaling suppress tumor growth and homologous recombination in aerodigestive cancers. Am J Cancer Res 5:1337–1352PubMedPubMedCentral

42.

Brechbiel J, Miller-Moslin K, Adjei AA (2014) Crosstalk between hedgehog and other signaling pathways as a basis for combination therapies in cancer. Cancer Treat Rev 40:750–759. doi:10.1016/j.ctrv.2014.02.003 CrossRefPubMed

43.

Calabrese EJ, Nascarella MA (2010) Tumor resistance explained by hormesis. Dose Response 8:80–82. doi:10.2203/dose-response.09-063.Calabrese CrossRefPubMedPubMedCentral

44.

Graham RA, Lum BL, Cheeti S et al (2011) Pharmacokinetics of hedgehog pathway inhibitor vismodegib (GDC-0449) in patients with locally advanced or metastatic solid tumors: the role of alpha-1-acid glycoprotein binding. Clin Cancer Res 17:2512–2520. doi:10.1158/1078-0432.CCR-10-2736 CrossRefPubMedPubMedCentral

45.

Gajjar AJ, Gururangan S, Qaddoumi IA et al (2013) A prospective phase II study to determine the efficacy of GDC 0449 (vismodegib) in adults with recurrent medulloblastoma (MB): a Pediatric Brain Tumor Consortium study (PBTC 25B). J Clin Oncol 31:suppl; abstr 2035

46.

Berlin J, Bendell JC, Hart LL et al (2013) A randomized phase II trial of vismodegib versus placebo with FOLFOX or FOLFIRI and bevacizumab in patients with previously untreated metastatic colorectal cancer. Clin Cancer Res 19:258–267. doi:10.1158/1078-0432.CCR-12-1800 CrossRefPubMed

47.

Kaye SB, Fehrenbacher L, Holloway R et al (2012) A phase II, randomized, placebo-controlled study of vismodegib as maintenance therapy in patients with ovarian cancer in second or third complete remission. Clin Cancer Res 18:6509–6518. doi:10.1158/1078-0432.CCR-12-1796 CrossRefPubMed

48.

Kim EJ, Sahai V, Abel EV et al (2014) Pilot clinical trial of hedgehog pathway inhibitor GDC-0449 (vismodegib) in combination with gemcitabine in patients with metastatic pancreatic adenocarcinoma. Clin Cancer Res 20:5937–5945. doi:10.1158/1078-0432.CCR-14-1269 CrossRefPubMedPubMedCentral

Title: Reduced proliferation and colony formation of head and neck squamous cell carcinoma (HNSCC) after dual targeting of EGFR and hedgehog pathways
Authors: Hannes Liebig
Georg Günther
Marlen Kolb
Christian Mozet
Andreas Boehm
Andreas Dietz
Gunnar Wichmann
Publication date: 01-02-2017
Publisher: Springer Berlin Heidelberg
Published in: Cancer Chemotherapy and Pharmacology / Issue 2/2017
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-017-3239-3

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Purpose

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