Reduced proliferation and colony formation of head and neck squamous cell carcinoma (HNSCC) after dual targeting of EGFR and hedgehog pathways
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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.
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.
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.
We provide ex-vivo evidence for vismodegib’s potential in HNSCC therapies, especially if combined with cetuximab, cisplatin and docetaxel.
KeywordsHead and neck squamous cell carcinoma (HNSCC) Head and neck cancer (HNC) Vismodegib Erivedge® Hedgehog signalling pathway SMO
We thank Anett Reiche for excellent technical assistance and the patients and their families for participation in the study.
Compliance with ethical standards
The study was supported by the Grant EPGZ268406-G from Roche Pharma AG.
Conflict of interest
Gunnar Wichmann received funding for a translational research project on “Efficacy of vismodegib (GDC-0449) on HNSCC ex vivo, EPGZ268406-G” from Roche Pharma AG. All other authors declare no conflict of interest.
The study was performed in accordance with the ethical standards as laid down in the 1964 declaration of Helsinki and its later amendments (vote 201-10-12072010 of the local ethics committee at the Medical Faculty of the University Leipzig).
- 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 CrossRefPubMedPubMedCentralGoogle Scholar
- 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 CrossRefPubMedPubMedCentralGoogle Scholar
- 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 2015Google Scholar
- 31.Sobin LH, Gospodarowicz MK, Wittekind C (2010) TNM classification of malignant tumours. Wiley, OxfordGoogle Scholar
- 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 CrossRefPubMedPubMedCentralGoogle Scholar
- 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 CrossRefPubMedGoogle Scholar
- 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 CrossRefPubMedPubMedCentralGoogle Scholar
- 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 2035Google Scholar
- 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 CrossRefPubMedPubMedCentralGoogle Scholar