In vivo Wnt pathway inhibition of human squamous cell carcinoma growth and metastasis in the chick chorioallantoic model.
BACKGROUND
Head and neck squamous cell carcinoma (HNSCC) is an aggressive cancer with poor overall survival. New therapeutic strategies that target specific molecular lesions driving advanced disease are needed. Herein we demonstrate the utility of the chicken chorioallantoic membrane (CAM) assay for in vivo human HNSCC tumor growth and metastasis and the tumor suppressive effects of a new chemotherapeutic agent.


METHODS
We tested anti-metastatic effects of a WNT pathway inhibitor, WNT974 (also known as LGK974), which targets porcupine (PORCN) the palmityl-transferase that is essential for secretion of Wnt proteins. CAM assays were performed with 8 HNSCC cell lines: UM-SCC-1, UM-SCC-10A, UM-SCC-10B, UM-SCC-11A, UM-SCC-14A UM-SCC-17A, UM-SCC-17B, UM-SCC-25, and UM-SCC-34.


RESULTS
UM-SCC-1 (University of Michigan Squamous Cell Carcinoma cell line) CAM xenografts contain CD44+ and ALDH+ cancer stem cell (CSC) proportions similar to UM-SCC-1 mouse xenografts supporting the applicability of the CAM assay for study of CSCs. Inhibition of WNT signaling by the PORCN inhibitor WNT974 reduced metastatic spread of UM-SCC cells, especially in UM-SCCs with Notch1 deficiency.


CONCLUSIONS
Our data demonstrate decreased tumor growth and metastases in tumors from cell lines that showed in vitro responses to WNT974, providing evidence that this agent may have a role in future HNSCC therapy.
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.|Department of Otolaryngology/Head Neck Surgery, Stanford University, Stanford, CA, 94305, USA.
Shannon F
SF
Rudy
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.|Comprehensive Cancer Center, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
J Chad
JC
Brenner
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA, 92121, USA.
Jennifer L
JL
Harris
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA, 92121, USA.
Jun
J
Liu
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA, 92121, USA.
Jianwei
J
Che
Comprehensive Cancer Center, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.|Wayne State University School of Medicine, Detroit, MI, 48201, USA.
Megan V
MV
Scott
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
John Henry
JH
Owen
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
Christine M
CM
Komarck
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
Martin P
MP
Graham
Cancer Center Biostatistics Core, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
Emily L
EL
Bellile
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.|Comprehensive Cancer Center, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
Carol R
CR
Bradford
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.|Comprehensive Cancer Center, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA.
Mark E P
ME
Prince
Department of Otolaryngology/Head Neck Surgery, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA. careyte@med.umich.edu.|Comprehensive Cancer Center, University of Michigan School of Medicine, Ann Arbor, MI, 48109-5616, USA. careyte@med.umich.edu.
Thomas E
TE
Carey
D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000378:metabolism / Q000556:secondary; D045744:Cell Line, Tumor; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000378:metabolism* / Q000473:pathology; D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis; D006258:Head and Neck Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000556:secondary; D006801:Humans; D051379:Mice; D009154:Mutation; D009362:Neoplasm Metastasis; D009374:Neoplasms, Experimental; D016133:Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000235:genetics* / Q000378:metabolism; D000077195:Squamous Cell Carcinoma of Head and Neck; D051153:Wnt Proteins / Q000235:genetics* / Q000378:metabolism; D060449:Wnt Signaling Pathway
D051881:Receptor, Notch1; C110670:WNT9A protein, human; D051153:Wnt Proteins; D004247:DNA
Cell lines; Chorioallantoic membrane; Human squamous cell carcinoma; In vivo cancer model; NOTCH1 mutation; UM-SCC; WNT pathway inhibition; WNT974