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Malignant pleural mesothelioma (MPM) is locally aggressive and challengingto quantitate non-invasively in vivo, particularly in orthotopic models of disease.We describe imaging of extracellular protease activity, typically elevated inlocally aggressive tumors, as a novel method for tracking MPM in vivo. Mice bearinghuman MPM subcutaneous flank xenografted tumors were imaged with ProSense 680,an optical imaging agent of extracellular cysteine protease activity. The relativecontribution of extracellular cysteine proteases to the ProSense tumor signalwas estimated using RT-PCR quantitation of cysteine protease RNA expression ofthe MPM cell lines and compared to ArrayExpress microarray RNA expression datafrom human MPM tumors. Feasibility of orthotopic intraperitoneal MPM cell masstracking with fluorescence signal was evaluated using CellVue Maroon-coated MSTO-211Hand compared to bioluminescent signal using luciferase-transfected MSTO-211H cells.ProSense 680 yielded a robust tumor signal in MPM subcutaneous grafts, primarilyresulting from MPM secretion of cathepsin L demonstrated not only by RT-PCR dataon MPM cell lines but also by microarray expression data from resected human patienttumors. CellVue Maroon intraperitoneal tumor signal was robust and durable indicatingfeasibility of intraperitoneal cell mass tracking of orthotopically-xenograftedMPM. Optical imaging of extracellular cysteine protease activity is useful fortracking MPM tumor cell mass in vivo. Intraperitoneal MPM cell mass tracking offluorescently labeled tumor is feasible.

Extracellular protease imaging for cell mass tracking of xenografted human malignant pleural mesothelioma