Abstract
Pancreatic cancer is generally detected at later stages with a poor prognosis and a high-mortality rate. Development of theranostic imaging agents that noninvasively target pancreatic cancer by gene expression and deliver therapies directly to malignant cells could greatly improve therapeutic outcomes. Small-peptide ligands that bind cell-surface proteins and are conjugated to imaging moieties have demonstrated efficacy in cancer imaging. Identification of cancer-specific targets is a major bottleneck in the development of such agents. Herein, a method is presented that uses DNA microarray expression profiling of large sets of normal and cancer tissues to identify targets expressed in cancer but not expressed in relevant normal tissues. Identified targets are subsequently validated for protein expression using tissue microarray. Further validations are performed by quantifying expression in pancreatic cancer cells by quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR), by immunocytochemistry and immunohistochemistry, and by reviewing data and literature in public databases. Validated targets are selected for ligand development based on the existence of a known ligand or by known structure–activity relationships useful for development of novel ligands.
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Dunphy, E. P. (2008) Pancreatic cancer: a review and update. Clin J Oncol Nurs 12, 735–741.
Klapman, J. and Malafa, M. P. (2008) Early detection of pancreatic cancer: why, who, and how to screen. Cancer Control 15, 280–287.
Goldenberg, D. M. and Sharkey, R. M. (2007) Novel radiolabeled antibody conjugates. Oncogene 26, 3734–3744.
Jhanwar, Y. S. and Divgi, C. (2005) Current status of therapy of solid tumors. J Nucl Med 46 Suppl 1, 141–150.
Goldenberg, D. M. and Sharkey, R. M. (2006) Advances in cancer therapy with radiolabeled monoclonal antibodies. Q J Nucl Med Mol Imaging 50, 248–264.
Temming, K., Schiffelers, R. M., Molema, G., and Kok, R. J. (2005) RGD-based strategies for selective delivery of therapeutics and imaging agents to the tumour vasculature. Drug Resist Updat 8, 381–402.
Haubner, R., Weber, W. A., Beer, A. J., et al. (2005) Noninvasive visualization of the activated avh3 integrin in cancer patients by positron emission tomography and [18F]galacto-RGD. PLoS Med 2, e70.
Beer, A. J., Haubner, R., Wolf, I., et al. (2006) PET-based human dosimetry of 18F-galacto-RGD, a new radiotracer for imaging avh3 expression. J Nucl Med 47, 763–769.
Fung, S. and Hruby, V. J. (2005) Design of cyclic and other templates for potent and selective peptide α-MSH analogues. Curr Opin Chem Biol 9, 352–358.
Handl, H. L., Vagner, J., Yamamura, H. I., Hruby, V. J., and Gillies, R. J. (2005) Development of a lanthanide-based assay for detection of receptor–ligand interactions at the delta-opioid receptor. Anal Biochem 343, 299–307.
Black, K. C., Kirkpatrick, N. D., Troutman, T. S., Xu, L., Vagner, J., Gillies, R. J., Barton, J. K., Utzinger, U., and Romanowski, M. (2008) Gold nanorods targeted to delta opioid receptor: plasmon-resonant contrast and photothermal agents. Mol Imaging 7, 50–57.
Balagurunathan, Y., Morse, D. L., Hostetter, G., Shanmugam, V., Stafford, P., et al. (2008) Gene expression profiling-based identification of cell-surface targets for developing multimeric ligands in pancreatic cancer. Mol Cancer Ther 7, 3071–3080.
Tangrea, M. A., Wallis, B. S., Gillespie, J. W., Gannot, G., Emmert-Buck, M. R., and Chuaqui, R. F. (2004) Novel proteomic approaches for tissue analysis. Expert Rev Proteomics 1, 185–192.
Kononen, J., Bubendorf, L., Kallioniemi, A., et al. (1998) Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med 4, 844–847.
Andersen, C. L., Hostetter, G., Grigoryan, A., Sauter, G., and Kallioniemi, A. (2001) Improved procedure for fluorescence in situ hybridization on tissue microarrays. Cytometry 45, 83–86.
Mousses, S., Bubendorf, L., Wagner, U., et al. (2002) Clinical validation of candidate genes associated with prostate cancer progression in the CWR22 model system using tissue microarrays. Cancer Res 62, 1256–1260.
Watanabe, A., Cornelison, R., and Hostetter, G. (2005) Tissue microarrays: applications in genomic research. Expert Rev Mol Diagn 5, 171–181.
Morse, D. L., Carroll, D., Weberg, L., Borgstrom, M. C., Ranger-Moore, J., and Gillies, R. J. (2005) Determining suitable internal standards for mRNA quantification of increasing cancer progression in human breast cells by real-time reverse transcriptase polymerase chain reaction. Anal Biochem 342, 69–77.
Lynch, R. M., Fogarty, K. E., and Fay, F. S. (1991) Modulation of hexokinase association with mitochondria analyzed with quantitative three-dimensional confocal microscopy. J Cell Biol 112, 385–395.
Bittner, M., Meltzer, P., Chen, Y., Jiang, Y., Seftor, E., Hendrix, M., Radmacher, M., Simon, R., Yakhinik, Z., Ben-Dork, A., Sampask, N., Dougherty, E., Wang, E., Marincola, F., Gooden, C., Lueders, J., Glatfelter, A., Pollock, P., Carpten, J., Gillanders, E., Leja, D., Dietrich, K., Beaudry, C., Berens, M., Alberts, D., Sondak, V., Hayward, N., and Trent, J. (2000) Molecular classification of cutaneous malignant melanoma by gene expression profiling. Nat Lett 406, 536–540.
Acknowledgments
We would like to thank Dr. Michael Bittner and Sonsoles Shack for microarray data acquisition, and Dr. Ronald Lynch for immunocytochemistry advice.
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Morse, D.L., Hostetter, G., Balagurunathan, Y., Gillies, R.J., Han, H. (2010). Identification of Pancreatic Cancer-Specific Cell-Surface Markers for Development of Targeting Ligands. In: Grobmyer, S., Moudgil, B. (eds) Cancer Nanotechnology. Methods in Molecular Biology, vol 624. Humana Press. https://doi.org/10.1007/978-1-60761-609-2_13
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DOI: https://doi.org/10.1007/978-1-60761-609-2_13
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