Matrix-metalloproteinases in head and neck carcinoma-cancer genome atlas analysis and fluorescence imaging in mice.

TitleMatrix-metalloproteinases in head and neck carcinoma-cancer genome atlas analysis and fluorescence imaging in mice.
Publication TypeJournal Article
Year of Publication2014
AuthorsHauff SJ, Raju SC, Orosco RK, Gross AM, Diaz-Perez JA, Savariar E, Nashi N, Hasselman J, Whitney M, Myers JN, Lippman SM, Tsien RY, Ideker T, Nguyen QT
JournalOtolaryngol Head Neck Surg
Date Published2014 Oct
KeywordsAnimals, Carcinoma, Squamous Cell, Cell Line, Tumor, Cell-Penetrating Peptides, Disease Models, Animal, Head and Neck Neoplasms, Humans, Matrix Metalloproteinases, Mice, Neoplasm Proteins, Neoplasm Transplantation, Optical Imaging, Papillomaviridae, Retrospective Studies, RNA, Messenger

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).

STUDY DESIGN: Retrospective human cohort study; prospective animal study.

SETTING: Translational research laboratory.

SUBJECTS AND 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.

PubMed URL
Alternate TitleOtolaryngol Head Neck Surg
PubMed ID25091190
PubMed Central IDPMC4469264
Grant ListP50 CA097007 / CA / NCI NIH HHS / United States
R01 CA158448 / CA / NCI NIH HHS / United States
R01 EB014929 / EB / NIBIB NIH HHS / United States
RR031979 / RR / NCRR NIH HHS / United States
Bioinformatics and Systems Biology