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64Cu-Labeled Aptamers for Tumor-Targeted Radionuclide Delivery

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RNA Interference and Cancer Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1974))

Abstract

Aptamers are a class of oligonucleotides with high binding affinity and specificity with their targets. Additionally, aptamers are nontoxic, very thermally stable, and able to reversibly undergo denaturation and have a small size. Cancer-related aptamers can be used for tumor-targeted drug delivery, such as to deliver diagnostic and therapeutic radionuclides to target cancers. We describe the process for preparing a 64Cu-labeled modified A10 aptamer to target prostate cancer by conjugating and radiolabeling. The modified A10 aptamer was conjugated with p-SCN-Bn-NOTA as the chelator. Following this, the aptamer can be radiolabeled with the 64Cu radioisotope. NOTA was selected as the chelator of choice due to its commercial availability and widely demonstrated in vivo stability with the 64Cu radioisotope. Using this system, prostate cancer could potentially be targeted for noninvasive imaging using positron emission tomography (PET). Closely following this protocol allows many aptamers to be successfully radiolabeled to accurately and quantitatively trace their distribution in vivo for a wide range of medical applications.

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References

  1. Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249(4968):505–510. https://doi.org/10.1126/science.2200121

    Article  CAS  PubMed  Google Scholar 

  2. Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346:818. https://doi.org/10.1038/346818a0

    Article  CAS  Google Scholar 

  3. Yang J, Bowser MT (2013) Capillary electrophoresis–SELEX selection of catalytic DNA Aptamers for a small-molecule porphyrin target. Anal Chem 85(3):1525–1530. https://doi.org/10.1021/ac302721j

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. He X, Zhao Y, He D, Wang K, Xu F, Tang J (2012) ATP-responsive controlled release system using aptamer-functionalized mesoporous silica nanoparticles. Langmuir 28(35):12909–12915. https://doi.org/10.1021/la302767b

    Article  CAS  PubMed  Google Scholar 

  5. Bock LC, Griffin LC, Latham JA, Vermaas EH, Toole JJ (1992) Selection of single-stranded DNA molecules that bind and inhibit human thrombin. Nature 355:564. https://doi.org/10.1038/355564a0

    Article  CAS  Google Scholar 

  6. Li N, Nguyen HH, Byrom M, Ellington AD (2011) Inhibition of cell proliferation by an anti-EGFR Aptamer. PLoS One 6(6):e20299. https://doi.org/10.1371/journal.pone.0020299

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Srisawat C, Goldstein IJ, Engelke DR (2001) Sephadex-binding RNA ligands: rapid affinity purification of RNA from complex RNA mixtures. Nucleic Acids Res 29(2):e4–e4. https://doi.org/10.1093/nar/29.2.e4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Jeong S, Eom T-Y, Kim S-J, Lee S-W, Yu J (2001) In vitro selection of the RNA aptamer against the Sialyl Lewis X and its inhibition of the cell adhesion. Biochem Biophys Res Commun 281(1):237–243. https://doi.org/10.1006/bbrc.2001.4327

    Article  CAS  PubMed  Google Scholar 

  9. Esposito CL, Cerchia L, Catuogno S, De Vita G, Dassie JP, Santamaria G, Swiderski P, Condorelli G, Giangrande PH, de Franciscis V (2014) Multifunctional aptamer-miRNA conjugates for targeted cancer therapy. Mol Ther 22(6):1151–1163. https://doi.org/10.1038/mt.2014.5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Daniels DA, Chen H, Hicke BJ, Swiderek KM, Gold L (2003) A tenascin-C aptamer identified by tumor cell SELEX: systematic evolution of ligands by exponential enrichment. Proc Natl Acad Sci U S A 100(26):15416–15421. https://doi.org/10.1073/pnas.2136683100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Jayasena SD (1999) Aptamers: an emerging class of molecules that rival antibodies in diagnostics. Clin Chem 45(9):1628–1650

    CAS  PubMed  Google Scholar 

  12. Bouchard PR, Hutabarat RM, Thompson KM (2010) Discovery and development of therapeutic aptamers. Annu Rev Pharmacol Toxicol 50(1):237–257. https://doi.org/10.1146/annurev.pharmtox.010909.105547

    Article  CAS  PubMed  Google Scholar 

  13. Lee JF, Stovall GM, Ellington AD (2006) Aptamer therapeutics advance. Curr Opin Chem Biol 10(3):282–289. https://doi.org/10.1016/j.cbpa.2006.03.015

    Article  CAS  PubMed  Google Scholar 

  14. Fine SL, Martin DF, Kirkpatrick P (2005) Pegaptanib sodium. Nat Rev Drug Discov 4:187. https://doi.org/10.1038/nrd1677

    Article  CAS  PubMed  Google Scholar 

  15. US National Library of Medicine (2017) clinicaltrials.gov. https://www.clinicaltrials.gov/ct2/show/NCT03168139?term=NCT03168139&rank=1. Accessed 2 Dec 2017

  16. US National Library of Medicine (2014) clinicaltrials.gov. https://www.clinicaltrials.gov/ct2/show/NCT01547897?term=NCT01547897&rank=1. Accessed 2 Dec 2017

  17. US National Library of Medicine (2017) clinicaltrials.gov. https://www.clinicaltrials.gov/ct2/show/NCT03364153?term=NCT03364153&rank=1. Accessed 2 Dec 2017

  18. Pei X, Zhang J, Liu J (2014) Clinical applications of nucleic acid aptamers in cancer. Mol Clin Oncol 2(3):341–348. https://doi.org/10.3892/mco.2014.255

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Yazdian-Robati R, Ramezani M, Jalalian SH, Abnous K, Taghdisi SM (2016) Targeted delivery of Epirubicin to cancer cells by polyvalent aptamer system in vitro and in vivo. Pharm Res 33(9):2289–2297. https://doi.org/10.1007/s11095-016-1967-4

    Article  CAS  PubMed  Google Scholar 

  20. Li J, Zheng C, Cansiz S, Wu C, Xu J, Cui C, Liu Y, Hou W, Wang Y, Zhang L, Teng I, Yang H-H, Tan W (2015) Self-assembly of DNA nanohydrogels with controllable size and stimuli-responsive property for targeted gene regulation therapy. J Am Chem Soc 137(4):1412–1415. https://doi.org/10.1021/ja512293f

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Leach J, Wang A, Ye K, Jin S (2016) A RNA-DNA hybrid aptamer for nanoparticle-based prostate tumor targeted drug delivery. Int J Mol Sci 17(3):380

    Article  PubMed  PubMed Central  Google Scholar 

  22. Taghavi S, HashemNia A, Mosaffa F, Askarian S, Abnous K, Ramezani M (2016) Preparation and evaluation of polyethylenimine-functionalized carbon nanotubes tagged with 5TR1 aptamer for targeted delivery of Bcl-xL shRNA into breast cancer cells. Colloids Surf B Biointerfaces 140(Supplement C):28–39. https://doi.org/10.1016/j.colsurfb.2015.12.021

    Article  CAS  PubMed  Google Scholar 

  23. Gijs M, Aerts A, Impens N, Baatout S, Luxen A (2016) Aptamers as radiopharmaceuticals for nuclear imaging and therapy. Nucl Med Biol 43(4):253–271. https://doi.org/10.1016/j.nucmedbio.2015.09.005

    Article  CAS  PubMed  Google Scholar 

  24. McNamara JO 2nd, Andrechek ER, Wang Y, Viles KD, Rempel RE, Gilboa E, Sullenger BA, Giangrande PH (2006) Cell type-specific delivery of siRNAs with aptamer-siRNA chimeras. Nat Biotechnol 24(8):1005–1015. https://doi.org/10.1038/nbt1223

    Article  CAS  PubMed  Google Scholar 

  25. Jiang D, Im H-J, Sun H, Valdovinos HF, England CG, Ehlerding EB, Nickles RJ, Lee DS, Cho SY, Huang P, Cai W (2017) Radiolabeled pertuzumab for imaging of human epidermal growth factor receptor 2 expression in ovarian cancer. Eur J Nucl Med Mol Imaging 44(8):1296–1305. https://doi.org/10.1007/s00259-017-3663-y

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Yang Y, Hernandez R, Rao J, Yin L, Qu Y, Wu J, England CG, Graves SA, Lewis CM, Wang P, Meyerand ME, Nickles RJ, X-w B, Cai W (2015) Targeting CD146 with a 64Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas. Proc Natl Acad Sci U S A 112(47):E6525–E6534. https://doi.org/10.1073/pnas.1502648112

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Dassie JP, Liu XY, Thomas GS, Whitaker RM, Thiel KW, Stockdale KR, Meyerholz DK, McCaffrey AP, McNamara JO 2nd, Giangrande PH (2009) Systemic administration of optimized aptamer-siRNA chimeras promotes regression of PSMA-expressing tumors. Nat Biotechnol 27(9):839–849. https://doi.org/10.1038/nbt.1560

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This work was supported by the University of Wisconsin–Madison, the National Institutes of Health (P30CA014520), the Beijing Nova Program (Z171100001117024), and the Beijing Capital Special Development Application Program (Z141107002514159).

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Authors and Affiliations

  1. Department of Nuclear Medicine, Peking University First Hospital, Beijing, China

    Lei Kang

  2. Department of Radiology, University of Wisconsin–Madison, Madison, WI, USA

    Zachary T. Rosenkrans & Weibo Cai

  3. Department of Medical Physics, University of Wisconsin–Madison, Madison, WI, USA

    Zachary T. Rosenkrans & Weibo Cai

Authors
  1. Lei Kang
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  2. Zachary T. Rosenkrans
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  3. Weibo Cai
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Corresponding authors

Correspondence to Lei Kang or Weibo Cai .

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Editors and Affiliations

  1. Cancer Biology, CSIR-Centre for Cellular and Molecular Biology, Hyderabad, Telangana, India

    Lekha Dinesh Kumar

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Kang, L., Rosenkrans, Z.T., Cai, W. (2019). 64Cu-Labeled Aptamers for Tumor-Targeted Radionuclide Delivery. In: Dinesh Kumar, L. (eds) RNA Interference and Cancer Therapy. Methods in Molecular Biology, vol 1974. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9220-1_17

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  • DOI: https://doi.org/10.1007/978-1-4939-9220-1_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9219-5

  • Online ISBN: 978-1-4939-9220-1

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