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Published OnlineFirst June 6, 2017; DOI: 10.1158/1541-7786.MCR-17-0164 Near-Infrared Photoimmunotherapy Targeting PSMA Figure 4. In vivo histologic fluorescence distribution and histologic NIR-PIT effect. A, The regimen of NIR-PIT and imaging is shown. B, Fluorescence images of resected PC3pip- luc tumors. White light images (top) and IR700 fluorescence image (bottom). High fluorescence intensity was shown in PC3pip-luc tumor 24 hours after injection of anti-PSMA-IR700, but the fluorescence decreased 24 hours after NIR-PIT. C, Differential interference contrast (DIC) and fluorescence microscopy images of PC3pip-luc tumor xenografts. High fluorescence intensity was shown in PC3pip-luc tumor 24 hours after injection of anti-PSMA-IR700, but the fluorescence decreased 24 hours after NIR-PIT. Scale bars, 100 mm. D, Resected tumor stained with H&E. A few scattered clusters of damaged tumor cells were seen within a background of diffuse cellular necrosis and microhemorrhage after NIR-PIT, while no obvious damage was observed after anti-PSMA-IR700 alone with NIR light exposure. White scale bars, 100 mm; black scale bars, 20 mm. fluorescence signal of the tumor decreased due to dying cells and partial photobleaching, while the IR700 fluorescence gradually decreased over the following days in tumors receiv- ing anti-PSMA-IR700 but not NIR light exposure (Fig. 3B). NIR-PIT resulted in decreases in bioluminescence (Fig. 3C). Luciferase activity significantly decreased after NIR-PIT (P < 0.0001 vs. other control groups; Fig. 3D). In contrast, lucifer- ase activity of tumor in other control groups showed an increase due to rapid tumor growth. Tumor growth was sig- nificantly inhibited in the NIR-PIT treatment group compared with the other control groups (P < 0.001; Fig. 3E), and significantly prolonged survival was achieved in the NIR-PIT group (P < 0.0001 vs. other control groups; Fig. 3F). Surpris- ingly, more than two thirds of tumors were cured with this regimen of NIR-PIT. No significant therapeutic effect was observed in the control groups, including those receiving anti-PSMA-IR700 i.v. only or in mice receiving NIR light www.aacrjournals.org exposure only. There was no skin necrosis or toxicity attrib- utable to the anti-PSMA-IR700 in any group. Histological analysis The treatment and imaging regimen is shown in Fig. 4A. High fluorescence intensity was shown in tumors 24 hours after anti- PSMA-IR700 injection compared with that in control tumors. The majority of fluorescence signal in tumors disappeared 24 hours after NIR-PIT in resected tumor (Fig. 4B). In frozen histologic specimens, high fluorescence intensity was also shown in tumors 24 hours after anti-PSMA-IR700 injection, and the signal decreased 24 hours after NIR-PIT (Fig. 4C). H&E staining of NIR-PIT–treated PC3pip-luc tumors revealed diffuse necrosis and microhemorrhage, with scattered clusters of live but damaged tumor cells, while no obvious damage was observed in the tumor receiving only anti-PSMA-IR700 but no light (Fig. 4D). Mol Cancer Res; 15(9) September 2017 1159 Downloaded from mcr.aacrjournals.org on December 1, 2020. © 2017 American Association for Cancer Research.PDF Image | Near-Infrared Photoimmunotherapy Targeting Prostate Cancer
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