title: Fluorescent imaging of superficial head and neck squamous cell carcinoma using a γ-glutamyltranspeptidase-activated targeting agent: a pilot study.
abstract: Detecting superficial head and neck squamous cell carcinoma (HNSCC) by endoscopy is challenging because of limited morphological hallmarks, and iodine cannot be applied to head and neck lesions due to severe mucosal irritation. γ-glutamyltranspeptidase (GGT), a cell surface enzyme, is overexpressed in several cancers, and it has been reported that γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG), a fluorescent targeting agent which can be enzymatically activated and becomes fluorescent after cleavage of a GGT-specific sequence, can be activated within a few minutes after application to animal models. We investigated whether early HNSCC can be detected by applying gGlu-HMRG to clinical samples.
  
  
  
  gGlu-HMRG was applied to four HNSCC cell lines, and fluorescence was observed by fluorescence microscopy and flow cytometry. Immunohistological examination was performed in three recent cases of endoscopic submucosal dissection (ESD) to investigate GGT expression. Fluorescence imaging with gGlu-HMRG in eight clinical samples resected by ESD or surgery was performed, and fluorescence intensity of tumor and normal mucosa regions of interest (ROI) was prospectively measured.
  
  
  
  All four gGlu-HMRG-applied cell lines emitted green fluorescence. Immunohistological examination demonstrated that GGT was highly expressed in HNSCC of the recent three ESD cases but barely in the normal mucosa. Fluorescence imaging showed that iodine-voiding lesions became fluorescent within a few minutes after application of gGlu-HMRG in all eight resected tumors. Tumor ROI fluorescence intensity was significantly higher than in the normal mucosa five minutes after gGlu-HMRG application.
  
  
  
  Fluorescence imaging with gGlu-HMRG would be useful for early detection of HNSCC.
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Mizushima
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan. sonishi@pop.med.hokudai.ac.jp.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Ohnishi
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Shimizu
authors.affiliation: Department of Surgical Pathology, Hokkaido University Hospital, N14, W5, Kita-ku, Sapporo, 060-8648, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hatanaka
authors.affiliation: Department of Surgical Pathology, Hokkaido University Hospital, N14, W5, Kita-ku, Sapporo, 060-8648, Japan.
authors.forename: Kanako C
authors.initials: KC
authors.lastname: Hatanaka
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Hosono
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Kubota
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Mitsuteru
authors.initials: M
authors.lastname: Natsuizaka
authors.affiliation: Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
authors.forename: Mako
authors.initials: M
authors.lastname: Kamiya
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, N14, W5, Kita-ku, Sapporo, 060-8648, Japan.
authors.forename: Shouko
authors.initials: S
authors.lastname: Ono
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Homma
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, N14, W5, Kita-ku, Sapporo, 060-8648, Japan.
authors.forename: Mototsugu
authors.initials: M
authors.lastname: Kato
authors.affiliation: Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo, 060-8638, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Sakamoto
authors.affiliation: Laboratory of Chemical Biology and Molecular Imaging, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.|Japan Agency for Medical Research and Development (AMED)-CREST, 7-1 Ootemachi-1, Chiyoda-ku, Tokyo, 100-0004, Japan.
authors.forename: Yasuteru
authors.initials: Y
authors.lastname: Urano
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000000981:diagnostic imaging* / Q000201:enzymology* / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D006258:Head and Neck Neoplasms / Q000000981:diagnostic imaging* / Q000201:enzymology* / Q000473:pathology; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D010865:Pilot Projects; D011446:Prospective Studies; D000077195:Squamous Cell Carcinoma of Head and Neck; D005723:gamma-Glutamyltransferase / Q000378:metabolism*
chemical_list: D005723:gamma-Glutamyltransferase
keywords: Fluorescent imaging; Head and neck squamous cell carcinoma; γ-Glutamyltranspeptidase