Summary
Aptamers are in vitro evolved molecules that bind to target proteins with high affinity and specificity by adapting three-dimensional structures upon binding. Because cancer cells exhibit the activation of signaling pathways that are not usually activated in normal cells, RNA aptamers against such a cancer cell-specific signal can be useful lead molecules for cancer gene therapy. The Wnt/β-catenin signaling pathway plays important roles in a critical initiating event in the formation of various human cancers. Because mutations in β-catenin have been found to be responsible for human tumorigenesis, β-catenin is the molecular target for effective anticancer therapies. Here, we describe the selection of RNA aptamers against β-catenin/transcription factor (TCF) proteins and their intracellular expression as intramers. The RNA aptamers acted as central inhibitory players for multiple oncogenic functions of β-catenin in colon cancer cells. These data provide the proof-of-principle for the use of RNA aptamers for an effective anticancer gene therapy.
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References
Tuerk, C. and Gold, L. (1990) Systemic evolution of ligands by exponential enrichment: RNA lignads to bacteriophage T4 DNA polymerase. Science 249, 505–510.
Ellington, A. D. and Szostak, J. W. (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346, 818–822.
Robertson, D. L. and Joyce, G. F. (1990) Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA. Nat. Struct. Biol. 344, 467–468.
Bunka, D. H. and Stockley, P. G. (2006) Aptamers come of age - at last. Nat. Rev. Microbiol. 4, 588–596.
Lee, J. F., Stovall, G. M. and Ellington, A. D. (2006) Aptamer therapeutics advance. Curr. Opin. Chem. Biol. 10, 282–289.
Que-Gewirth, N. S. and Sullenger, B. A. (2007) Gene therapy progress and prospects: RNA aptamers. Gene Therapy 14, 283–291.
Farokhzad, O. C., Cheng, J., Teply, B. A., Sherifi, I., Jon, S., Kantoff, P. W., Richie, J. P., and Langer, R. (2006) Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. Proc. Natl. Acad. Sci. U.S.A. 103, 6315–6320.
Proske, D., Blank, M., Buhmann, R., and Resch, A. (2005) Aptamers-basic research, drug development, and clinical applications.Appl. Microbiol. Biotechnol. 69, 367–374.
Chu, T. C., Marks, J. W. III, lavery, L, A., Faulkner, S., Rosenblum, M. G., Ellington, A. D., and Levy, M. (2006) Aptamer:Toxin conjugates that specifically target prostate tumor cells. Cancer Res. 66, 5989–5992.
Polakis, P. (2000) Wnt signaling and cancer. Genes Dev. 14, 1837–1851.
Huelsken, J. and Birchmeier, W. (2001) New aspects of Wnt signaling pathways in higher vertebrates. Curr. Opin. Genet. Dev. 11, 547–553.
Gregorieff, A. and Clevers, H. (2005) Wnt signaling in the intestinal epithelium: from endoderm to cancer. Genes Dev. 19, 877–890.
Tetsu, O. and McCormick, F. (1999) β:-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature 398, 422–426.
He, T. C., Sparks, A. B., Rago, C., Hermeking, H., Zawel, L., da Costa, L. T., Morin, P. J., Vogelstein, B., and Kinzler, K. W. (1998) Identification of c-MYC as a target of the APC pathway. Science 281, 1509–1512.
Sato, S., Idogawa, M., Honda, K., Fujii, G., Kawashima, H., Takekuma, K., Hoshika, A., Hirohashi, S., and Yamada, T. (2005) β-Catenin interacts with the FUS proto-oncogene product and regulates pre mRNA splicing. Gastroenterology 129, 1225–1236.
Lee, H. K., Choi, Y. S., Park, Y. A., and Jeong, S. (2006) Modulation of oncogenic transcription and alternative splicing by β-catenin and an RNA aptamer in colon cancer cells. Cancer Res. 66, 10560–10566.
Lee, H. K. and Jeong, S. (2006) β:-Catenin stabilizes Cyclooxygenase-2 mRNA by interacting with AU-rich elements of 3'-UTR. Nucleic Acids Res. 34, 5705–5714.
Lee, H. K., Kwak, H. Y., Hur, J., Kim, I. A., Yang, J. S., Park, M. W., Yu, J., and Jeong, S. (2007) β-Catenin regulates multiple steps of RNA metabolism as revealed by the RNA aptamer in colon cancer cells. Cancer Res. 67, 9315–9321.
Miller, D., Laber, D., Bates, P., Trent, J., Taft, B., and Kloecker, G.H. (2006) Extended phase I study of AS1411 in renal and non-small cell lung cancers. Ann. Oncol. 17, ix147–148.
Famulok, M. and Mayer, G. (2005) Intramers and aptamers: applications in protein-function analyses and potential for drug screening. Chem. Bio. Chem. 6, 19–26.
Choi, K. H., Park, M. W., Lee, S. Y., Jeon, M. Y., Kim, M. Y., Lee, H. K., Yu, J., J. W. (2006) In vitro selection of RNA molecules that bind specific ligands. Mol Cancer Ther 5, 1418–1434.
Acknowledgments
This study was supported by grants from the Korea Research Foundation, the Korea Science and Engineering Foundation, the Korean Ministry of Sciences and Technology, and the Korean Ministry of Health and Welfare.
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Jeong, S., Lee, H.K., Kim, M.Y. (2009). Use of RNA Aptamers for the Modulation of Cancer Cell Signaling. In: Walther, W., Stein, U. (eds) Gene Therapy of Cancer. Methods in Molecular Biology™, vol 542. Humana Press. https://doi.org/10.1007/978-1-59745-561-9_20
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DOI: https://doi.org/10.1007/978-1-59745-561-9_20
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