Skip to main content

Advertisement

SpringerLink
Log in

Selection of 2′F-modified RNA aptamers against prostate-specific antigen and their evaluation for diagnostic and therapeutic applications

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Currently, prostate-specific antigen (PSA) is considered to be the most sensitive marker available for prostate cancer detection and for monitoring of disease progression. In addition to its importance as a tumor marker, PSA has a role in the biological activity of cancer growth and proliferation. Therefore, the inhibition or activation of its biological activity may be used in prostate cancer therapy. Here, we describe the isolation and characterization of new 2′F-modified RNA aptamers directed against PSA. Binding studies demonstrate the ability of these new aptamers to specifically recognize their target with dissociation constants in the nanomolar range. In order to demonstrate the functionality of the selected aptamers, an apta-PCR approach was used for the quantitative detection of PSA, achieving a limit of detection of 11 nM. Furthermore, the potential use of the selected aptamers in therapeutics was demonstrated with the 2′F-modified aptamers being highly stable in human serum and having the ability to moderate the activity of PSA, which will be explored for the treatment of prostate cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Ferlay J, Parkin DM, Steliarova-Foucher E (2010) Estimates of cancer incidence and mortality in Europe in 2008. Eur J Cancer 46(4):765–781

    Article  CAS  Google Scholar 

  2. Jemal A, R. S, Ward E, Hao Y, Xu J, Thun MJ (2009) Cancer statistics. CA Cancer J Clin 59(4):225–249

    Article  Google Scholar 

  3. Siegel R, Ward E, Brawley O, Jemal A (2011) The impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA: A Cancer J Clin 61(4):212–236

    Article  Google Scholar 

  4. Lilja H, Oldbring J, Rannevik G, Laurell CB (1987) Seminal vesicle-secreted proteins and their reactions during gelation and liquefaction of human semen. J Clin Invest 80(2):281–285

    Article  CAS  Google Scholar 

  5. Thompson IM, Pauler DK, Goodman PJ, Tangen CM, Lucia M, Parnes HL, Minasian LM, Ford LG, Lippman SM, Crawford ED, Crowley JJ, Coltman CAJ (2004) Prevalence of prostate cancer among men with a prostate-specific antigen level < or = 4.0 ng per milliliter. N Engl J Med:2239–2246

  6. Cohen P, Graves HC, Pechl DM, Kamarei M, Guidice LC, Rosenfeld RG (1992) Prostate-specific antigen is an insulin-like growth factor binding protein-3 protease found in seminal plasma. J Clin Endocrin Metab 75:1046–1053

    Article  CAS  Google Scholar 

  7. Rehault S, Monget P, Mazerbourg S, Tremblaz R, Gutman N, Gauthir F, Moreau T (2001) Insulin-like growth factor binding protein (IGFBPs) as potential physiological substrates for human kallikreins hK2 and hK3. Eu J Biochem 268:2960–2968

    Article  CAS  Google Scholar 

  8. Killian CS, Corral DA, Kaminski E, Constantine RI (1993) Mitogenic response of osteoblast cell to prostate-specific antigen suggests an activation of latent TGF-beta and a proteolytic modulation of cell adhesion receptors. Biochem Bioph Res Comm 192:940–947

    Article  CAS  Google Scholar 

  9. Webber MM, Waghray A, Bello D (1995) Prostate-specific antigen, a serine protease, facilitates human prostate cancer cell invasion. Clin Cancer Res 1:1089–1094

    CAS  Google Scholar 

  10. Gallardo-Williams MT, Maronpot RR, Wine RN, Brunssen SH, Chapin RE (2003) Inhibition of the enzymatic activity of prostate-specific antigen by boric acid and 3- nitrophenyl boronic acid. The Prostate 54:44–49

    Article  Google Scholar 

  11. Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346(6287):818–822

    Article  CAS  Google Scholar 

  12. Robertson DL, Joyce GF (1990) Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA. Nature 344(6265):467–468

    Article  CAS  Google Scholar 

  13. Tuerk C, Gold L (1990) Systematic evolution of ligands by exponentional enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249:505–510

    Article  CAS  Google Scholar 

  14. James W (2000) Aptamers. Encycl Anal Chem:4848–4871

  15. Lupold SE, Hicke BJ, Lin Y, Coffey DS (2002) Identification and characterization of nuclease-stabilized RNA molecules that bind human prostate cancer cells via the prostate-specific membrane antigen. Cancer Res 62:4029–4033

    CAS  Google Scholar 

  16. Mayer G (2009) The chemical biology of aptamers. Angew Chem Int Ed 48:2672–2689

    Article  CAS  Google Scholar 

  17. Bagalkot V, Farokhzad OC, Langer R, Jon S (2006) An aptamer–doxorubicin physical conjugate as a novel targeted drug-delivery platform. Angew Chem Int Ed Engl 45(48):8149–8152

    Article  CAS  Google Scholar 

  18. Cheng J, Teply BA, Sherifi I, Sung J, Luther G, Gu FX, Levy-Nissenbaum E, Radovic-Moreno AF, Langer R, Farokhzad OC (2007) Formulation of functionalized PLGA–PEG nanoparticles for in vivo targeted drug delivery. Biomaterials 28(5):869–876

    Article  CAS  Google Scholar 

  19. Chu TC, Twu KY, Ellington AD, Levy M (2006) Aptamer mediated siRNA delivery. Nucleic Acid Res 34(10):e73

    Article  Google Scholar 

  20. Farokhzad OC, Cheng J, Teply BA, Sherifi I, Jon S, Kantoff PW, Richie JP, Langer R (2006) Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. Proc Natl Acad Sci U S A 103(16):6315–6320

    Article  CAS  Google Scholar 

  21. Farokhzad OC, Jon SY, Khademhosseini A, Tran TT, LaVan D, Langer R (2004) Nanoparticle–aptamer bioconjugates: a new approach for targeting prostate cancer cells. Cancer Res 64(21):7668–7672

    Article  CAS  Google Scholar 

  22. McNamara JO, 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

    Article  CAS  Google Scholar 

  23. Jeong S, Han SR, Lee YJ, Lee SW (2010) Selection of RNA aptamers specific to active prostate-specific antigen. Biotechnol Lett 32:379–385

    Article  CAS  Google Scholar 

  24. Savory N, Abe K, Sode K, Ikebukuro K (2010) Selection of DNA aptamer against prostate specific antigen using a genetic algorithm and application to sensing. Biosens Bioelectron 26(4):1386–1391

    Article  CAS  Google Scholar 

  25. Ulrich H, Trujillo CA, Nery AA, Alves JM, Majumder P, Resende RR, Martins AH (2006) DNA and RNA aptamers: from tools for basic research towards therapeutic applications. Comb Chem High Throughput Screen 9(8):619–632

    Article  CAS  Google Scholar 

  26. Thiel KW, Giangrande PH (2009) Therapeutic applications of DNA and RNA aptamers. Oligonucleotides 19(3):209–222

    Article  CAS  Google Scholar 

  27. Padilla R, Sousa R (2002) A Y639F/H784A T7 RNA polymerase double mutant displays superior properties for synthesizing RNAs with non–canonical NTPs. Nucleic Acids Res 30(24):e138

    Article  Google Scholar 

  28. Bunka DHJ, Mantle BJ, Morten IJ, Tennent GA, Radford SE, Stockley PG (2007) Production and characterisation of RNA aptamers specific for amyloid fibril epitopes. J Biol Chem:M703679200. doi:10.1074/jbc.M703679200

  29. Zuker M (2003) M-fold web server for nucleic acid folding and hybridization prediction. 31:3406–3415

  30. Pinto A, Bermudo Redondo MC, Cengiz OV, O’Sullivan CK (2009) Real-time apta-PCR 20,000-fold improvement in detection limit. Mol Biosyst 5:548–553

    Article  CAS  Google Scholar 

  31. Chen Z, Lei Y, Chen X, Wang Z, Liu J (2012) An aptamer based resonance light scattering assay of prostate specific antigen. Biosens Bioelectron 36(1):35–40

    Article  CAS  Google Scholar 

  32. Black MH, Grass LC, Leinonen J, Stenman U-H, Diamandis EP (1999) Characterization of monoclonal antibodies for prostate specific antigen and development of highly sensitive free prostate-specific antigen assays. Clin Chem 45(3):347–354

    CAS  Google Scholar 

  33. Niemelä P, Lövgren J, Karp M, Lilja H, Pettersson K (2002) Sensitive and specific enzymatic assay for the determination of precursor forms of prostate-specific antigen after an activation step. Clin Chem 48(8):1257–1264

    Google Scholar 

  34. Xie C, Wang G (2011) Development of simultaneous detection of total prostate-specific antigen (tPSA) and free PSA with rapid bead-based immunoassay. J Clin Lab Anal 25(1):37–42

    Article  CAS  Google Scholar 

  35. Osborne S, Ellington AD (1997) Nucleic acid selection and the challenge of combinatorial chemistry. Chem Rev 97(2)

  36. Kusser W (2000) Chemically modified nucleic acid aptamers for in vitro selections: evolving evolution. J Biotechnol 74(1):27–38

    CAS  Google Scholar 

  37. Adlington RM, Baldwin JE, Becker GW, Chen B, Cheng L, Cooper SL, Hermann RB, Howe TJ, McCoull W, McNulty AM, Neubauer BL, Pritchard GJ (2001) Design, synthesis, and proposed active site binding analysis of monocyclic 2-azetidinone inhibitors of prostate specific antigen. J Med Chem 44(10):1491–1508

    Article  CAS  Google Scholar 

  38. Annunziata R, Benaglia M, Cinquini M, Cozzi F, Puglisi A (2002) Efficient and highly stereoselective synthesis of a beta-lactam inhibitor of the serine protease prostate-specific antigen. Bioorg Med Chem 10(6):1813–1818

    Article  CAS  Google Scholar 

  39. Corey E, Wegner SK, Stray JE, Corey MJ, Arfman EW, Lange PH, Vessella RL (1997) Characterization of 10 new monoclonal antibodies against prostate-specific antigen by analysis of affinity, specificity and function in sandwich assays. Int J Cancer 71(6):1019–1028

    Article  CAS  Google Scholar 

  40. Ireson CR, Kelland LR (2006) Discovery and development of anticancer aptamers. Mol Cancer Ther 5(12):2957–2962

    Article  CAS  Google Scholar 

  41. Malm J, Hellman J, Hogg P, Lilja H (2000) Enzymatic action of prostatespecific antigen (PSA or hK3): substrate specificity and regulation by Zn(2+), a tight-binding inhibitor. The Prostate 45:132–139

    Article  CAS  Google Scholar 

Download references

Acknowledgments

M.S. acknowledges the support from a FI predoctoral scholarship of the Generalitat de Catalunya.

Author information

Authors and Affiliations

  1. Nanobiotechnology and Bioanalysis group, Department of Chemical Engineering, Universitat Rovira i Virgili, 43007, Tarragona, Spain

    M. Svobodova, P. Nadal & C. K. O’Sullivan

  2. Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK

    D. H. J. Bunka & P. G. Stockley

  3. Institució Catalana de Recerca i Estudis Avançats, Passeig Lluís Companys 23, 08010, Barcelona, Spain

    C. K. O’Sullivan

Authors
  1. M. Svobodova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. H. J. Bunka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. Nadal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. G. Stockley
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C. K. O’Sullivan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to M. Svobodova or C. K. O’Sullivan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Svobodova, M., Bunka, D.H.J., Nadal, P. et al. Selection of 2′F-modified RNA aptamers against prostate-specific antigen and their evaluation for diagnostic and therapeutic applications. Anal Bioanal Chem 405, 9149–9157 (2013). https://doi.org/10.1007/s00216-013-7350-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-013-7350-y

Keywords

Search

Navigation