title: Matrix-metalloproteinases in head and neck carcinoma-cancer genome atlas analysis and fluorescence imaging in mice.
abstract: OBJECTIVE
  (1) Obtain matrix-metalloproteinase (MMP) expression profiles for head and neck squamous cell carcinoma (HNSCC) specimens from the Cancer Genomic Atlas (TCGA). (2) Demonstrate HNSCC imaging using MMP-cleavable, fluorescently labeled ratiometric activatable cell-penetrating peptide (RACPP).
  
  
  METHODS
  Retrospective human cohort study; prospective animal study.
  
  
  METHODS
  Translational research laboratory.
  
  
  METHODS
  Patient clinical data and mRNA expression levels of MMP genes were downloaded from TCGA data portal. RACPP provides complementary ratiometric fluorescent contrast (increased Cy5 and decreased Cy7 intensities) when cleaved by MMP2/9. HNSCC-tumor bearing mice were imaged in vivo after RACPP injection. Histology was evaluated by a pathologist blinded to experimental conditions. Zymography confirmed MMP-2/9 activity in xenografts. RACPP was applied to homogenized human HNSCC specimens, and ratiometric fluorescent signal was measured on a microplate reader for ex vivo analysis.
  
  
  RESULTS
  Expression of multiple MMPs including MMP2/9 is greater in patient HNSCC tumors than matched control tissue. In patients with human papilloma virus positive (HPV+) tumors, higher MMP2 and MMP14 expression correlates with worse 5-year survival. Orthotopic tongue HNSCC xenografts showed excellent ratiometric fluorescent labeling with MMP2/9-cleavable RACPP (sensitivity = 95.4%, specificity = 95.0%). Fluorescence ratios were greater in areas of higher tumor burden (P < .03), which is useful for intraoperative margin assessment. Ex vivo, human HNSCC specimens showed greater cleavage of RACPP when compared to control tissue (P = .009).
  
  
  CONCLUSIONS
  Human HNSCC tumors show increased mRNA expression of multiple MMPs including MMP2/9. We used RACPP, a ratiometric fluorescence assay of MMP2/9 activity, to show improved occult tumor identification and margin clearance. Ex vivo assays using RACPP in biopsy specimens may identify patients who will benefit from intraoperative RACPP use.
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA.
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Hauff
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA.
authors.forename: Sharat C
authors.initials: SC
authors.lastname: Raju
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA.
authors.forename: Ryan K
authors.initials: RK
authors.lastname: Orosco
authors.affiliation: Bioinformatics and Systems Biology Program, University of California, San Diego, California, USA.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Gross
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA.
authors.forename: Julio A
authors.initials: JA
authors.lastname: Diaz-Perez
authors.affiliation: Department of Pharmacology, University of California, San Diego, California, USA.
authors.forename: Elamprakash
authors.initials: E
authors.lastname: Savariar
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA.
authors.forename: Nadia
authors.initials: N
authors.lastname: Nashi
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Hasselman
authors.affiliation: Department of Pharmacology, University of California, San Diego, California, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Whitney
authors.affiliation: Department of Head and Neck Surgery, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
authors.forename: Jeffrey N
authors.initials: JN
authors.lastname: Myers
authors.affiliation: Moores Cancer Center, University of California, San Diego, California, USA.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Lippman
authors.affiliation: Department of Pharmacology, University of California, San Diego, California, USA Howard Hughes Medical Institute, San Diego, California, USA.
authors.forename: Roger Y
authors.initials: RY
authors.lastname: Tsien
authors.affiliation: Bioinformatics and Systems Biology Program, University of California, San Diego, California, USA Division of Medical Genetics, University of California, San Diego, California, USA.
authors.forename: Trey
authors.initials: T
authors.lastname: Ideker
authors.affiliation: Division of Head and Neck Surgery, University of California, San Diego, California, USA Moores Cancer Center, University of California, San Diego, California, USA quyennguyen@ucsd.edu.
authors.forename: Quyen T
authors.initials: QT
authors.lastname: Nguyen
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000201:enzymology* / Q000401:mortality / Q000821:virology; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides; D004195:Disease Models, Animal; D006258:Head and Neck Neoplasms / Q000201:enzymology* / Q000401:mortality / Q000821:virology; D006801:Humans; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D009368:Neoplasm Transplantation; D061848:Optical Imaging; D027383:Papillomaviridae; D012333:RNA, Messenger / Q000378:metabolism; D012189:Retrospective Studies; D000077195:Squamous Cell Carcinoma of Head and Neck
chemical_list: D057846:Cell-Penetrating Peptides; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D020782:Matrix Metalloproteinases
keywords: fluorescence imaging; head and neck squamous cell carcinoma; human papilloma virus; the Cancer Genomic Atlas
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