Abstract
Technologies are needed to map and image biological barriers in vivo that limit solid tumor delivery and, ultimately, the effectiveness of imaging and therapeutic agents. Here we integrate proteomic and imaging analyses of caveolae at the blood-tumor interface to discover an active transendothelial portal to infiltrate tumors. A post-translationally modified form of annexin A1 (AnnA1) is selectively concentrated in human and rodent tumor caveolae. To follow trafficking, we generated a specific AnnA1 antibody that targets caveolae in the tumor endothelium. Intravital microscopy of caveolae-immunotargeted fluorophores even at low intravenous doses showed rapid and robust pumping across the endothelium to enter mammary, prostate and lung tumors. Within 1 h, the fluorescence signal concentrated throughout tumors to exceed the peak levels in blood. This transvascular pumping required the expression of caveolin 1 and annexin A1. Tumor uptake with other antibodies were >100-fold less. This proteomic imaging strategy reveals a unique target, antibody and caveolae pumping system for solid tumor penetration.
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Acknowledgements
We thank R.D. Winger for performing small animal surgeries and R. Baldwin for assistance with gold labeling experiments. We also thank R.J. Flower (Queen Mary University of London), M. Huflejt (New York University), J. Lustgarten (Mayo Clinic), D. Sanger (Beth Israel Deaconess Medical Center) and R. Tsien (University of California San Diego) for their kind gifts. This research was supported in part by the Komen Foundation, the US Department of Defense (Prostate Cancer Research Program Award number W81XWH-11-1-0693), the state of California (Tobacco-Related Disease Research Program grant 18XT0196 and California Breast Cancer Research Program grant 16IB-0104) and the US National Institutes of Health (R01CA115215, P01CA104898, R01CA119378 and R01CA83989).
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P.O. performed tissue subcellular fractionations, tumor implantations, antibody conjugations, crosslinking studies and western blot analyses. J.E.T. cloned and expressed full-length annexin A1 proteins, performed western blot and immunohistochemical analyses and generated, characterized and supplied mAnnA1. H.W. performed EM analysis with P.O. P.O. and P.B. performed IVM imaging and quantitative analyses. Y.L. performed mass spectrometry analysis. P.O. and J.E.S. designed the experiments and wrote the manuscript.
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Supplementary Figures 1 and 2 and Supplementary Note (PDF 919 kb)
41591_2014_BFnm3623_MOESM14_ESM.mov
Continuous imaging of rapid solid tumor penetration with fluorophore linked to mAnnA1 when injected intravenously (MOV 72507 kb)
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Oh, P., Testa, J., Borgstrom, P. et al. In vivo proteomic imaging analysis of caveolae reveals pumping system to penetrate solid tumors. Nat Med 20, 1062–1068 (2014). https://doi.org/10.1038/nm.3623
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DOI: https://doi.org/10.1038/nm.3623
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