Advertisement

Molecular Medicine

, Volume 17, Issue 11–12, pp 1213–1222 | Cite as

Small-Interfering RNA-Eluting Surfaces as a Novel Concept for Intravascular Local Gene Silencing

  • Andrea Nolte
  • Tobias Walker
  • Martina Schneider
  • Oya Kray
  • Meltem Avci-Adali
  • Gerhard Ziemer
  • Hans Peter Wendel
Research Article

Abstract

New drug-eluting stent (DES) methods have recently been demonstrated to improve outcomes of intravascular interventions. A novel technique is the design of gene-silencing stents that elute specific small-interfering RNAs (siRNAs) for better vascular wall regeneration. Although siRNAs used to alter gene expression have surpassed expectations in in vitro experiments, the functional and local delivery of siRNAs is still the major obstacle for the in vivo application of RNA interference. In this preliminary in vitro study we investigated a surface-immobilized siRNA delivery technique that would be readily adaptable for local intravascular applications in vivo. The transfection potency of gelatin coatings consisting of a specific siRNA complexed with polyethylenimine (PEI) was examined in primary human endothelial cells by flow cytometry and quantitative real-time polymerase chain reaction. Several media conditions, such as the presence or absence of serum during cultivation, were investigated. Furthermore, different siRNA and PEI amounts, as well as nitrogen/phosphate ratios, were tested for their transfection efficiency. Gelatin coatings consisting of PEI and siRNA against an exemplary endothelial adhesion molecule receptor achieved a significant knockdown of around 70%. The transfection efficiency of the coatings was not influenced by the presence of serum. The results of this preliminary study support the expectation that this novel coating may be favorable for local in vivo gene silencing (for example, when immobilized on stents or balloons for percutanous transluminal coronary angioplasty). However, further animal experiments are needed to confirm the translation into clinical practice. This intriguing technology leads the way to more sophisticated and individualized coatings for the post-DES era, toward silencing of genes involved in the pathway of intimal hyperplasia.

References

  1. 1.
    Greenhalgh J, et al. (2010) Drug-eluting stents versus bare metal stents for angina or acute coronary syndromes. Cochrane Database Syst. Rev. 5:CD004587.Google Scholar
  2. 2.
    Wendel HP, Avci-Adali M, Ziemer G. (2010) Enothelial progenitor cell capture stents: hype or hope? Int. J. Cardiol. 5;145:115–7CrossRefGoogle Scholar
  3. 3.
    Dorsett Y, Tuschl T. (2004) siRNAs: applications in functional genomics and potential as therapeutics. Nat. Rev. Drug. Discov. 3:318–29.CrossRefGoogle Scholar
  4. 4.
    Elbashir SM, et al. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411:494–8.CrossRefGoogle Scholar
  5. 5.
    Fire A, et al. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391:806–811.CrossRefGoogle Scholar
  6. 6.
    Meister G, Tuschl T. (2004) Mechanisms of gene silencing by double-stranded RNA. Nature. 431:343–9.CrossRefGoogle Scholar
  7. 7.
    Chi JT, et al. (2003) Genomewide view of gene silencing by small interfering RNAs. Proc. Natl. Acad. Sci. U. S. A. 100:6343–6.CrossRefGoogle Scholar
  8. 8.
    Hassan A, et al. (2005) Small interfering RNA-mediated functional silencing of vasopressin V2 receptors in the mouse kidney. Physiol. Genomics. 21:382–8.CrossRefGoogle Scholar
  9. 9.
    Shimizu H, Fujita T. (2011) New short interfering RNA-based therapies for glomerulonephritis. Nat. Rev. Nephrol. 7:407–15CrossRefGoogle Scholar
  10. 10.
    Tagalakis AD, He L, Saraiva L, Gustafsson KT, Hart SL. (2011) Receptor-targeted liposomepeptide nanocomplexes for siRNA delivery. Biomaterials. 2:6302–15CrossRefGoogle Scholar
  11. 11.
    Kato M, et al. (2009) The targeting of cyclophilin D by RNAi as a novel cardioprotective therapy: evidence from two-photon imaging. Cardiovasc. Res. 83:335–44.CrossRefGoogle Scholar
  12. 12.
    Valentim L, et al. (2006) Urocortin inhibits Beclin1-mediated autophagic cell death in cardiac myocytes exposed to ischaemia/reperfusion injury. J. Mol. Cell. Cardiol. 40:846–52.CrossRefGoogle Scholar
  13. 13.
    Yin C, Xi L, Wang X, Eapen M, Kukreja RC. (2005) Silencing heat shock factor 1 by small interfering RNA abrogates heat shock-induced cardioprotection against ischemia-reperfusion injury in mice. J. Mol. Cell. Cardiol. 39:681–9.CrossRefGoogle Scholar
  14. 14.
    Zheng X, et al. (2009) Novel small interfering RNA-containing solution protecting donor or gans in heart transplantation. Circulation. 120:1099–107, 1091 p following 1107.CrossRefGoogle Scholar
  15. 15.
    Akhtar S, Benter IF. (2007) Nonviral delivery of synthetic siRNAs in vivo. J. Clin. Invest. 117:3623–32.CrossRefGoogle Scholar
  16. 16.
    Bumcrot D, Manoharan M, Koteliansky V, Sah DW. (2006) RNAi therapeutics: a potential new class of pharmaceutical drugs. Nat. Chem. Biol. 2:711–9.CrossRefGoogle Scholar
  17. 17.
    Lin X, et al. (2011) A robust in vivo positive-readout system for monitoring siRNA delivery to xenograft tumors. RNA. 17:603–12.CrossRefGoogle Scholar
  18. 18.
    Shim MS, Kwon YJ. (2010) Efficient and targeted delivery of siRNA in vivo. FEBS J. 277:4814–27.CrossRefGoogle Scholar
  19. 19.
    Walker T, et al. (2007) Inhibition of adhesion molecule expression on human venous endothelial cells by non-viral siRNA transfection. J. Cell. Mol. Med. 11:139–47.CrossRefGoogle Scholar
  20. 20.
    Collins T, et al. (1991) Structure and chromosomal location of the gene for endothelial-leukocyte adhesion molecule 1. J Biol. Chem. 266:2466–73.PubMedGoogle Scholar
  21. 21.
    Zhao QQ, et al. (2009) N/P ratio significantly influences the transfection efficiency and cytotoxicity of a polyethylenimine/chitosan/DNA complex. Biol. Pharm. Bull. 32:706–710.CrossRefGoogle Scholar
  22. 22.
    Leng Q, Woodle MC, Lu PY, Mixson AJ. (2009) Advances in systemic siRNA delivery. Drugs Future. 34:721.CrossRefGoogle Scholar
  23. 23.
    Werth S, et al. (2006) A low molecular weight fraction of polyethylenimine (PEI) displays increased transfection efficiency of DNA and siRNA in fresh or lyophilized complexes. J. Control Release. 112:257–70.CrossRefGoogle Scholar
  24. 24.
    Boussif O, et al. (1995) A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc. Natl. Acad. Sci. U. S. A. 92:7297–301.CrossRefGoogle Scholar
  25. 25.
    Finsinger D, Remy JS, Erbacher P, Koch C, Plank C. (2000) Protective copolymers for nonviral gene vectors: synthesis, vector characterization and application in gene delivery. Gene Ther. 7:1183–92.CrossRefGoogle Scholar
  26. 26.
    Berg MC, Zhai L, Cohen RE, Rubner MF. (2006) Controlled drug release from porous polyelectrolyte multilayers. Biomacromolecules 7:357–64.CrossRefGoogle Scholar
  27. 27.
    Thierry B, et al. (2005) Delivery platform for hydrophobic drugs: prodrug approach combined with self-assembled multilayers. J. Am. Chem. Soc. 127:1626–7.CrossRefGoogle Scholar
  28. 28.
    Thierry B, Winnik FM, Merhi Y, Silver J, Tabrizian M. (2003) Bioactive coatings of endovascular stents based on polyelectrolyte multilayers. Biomacromolecules. 4:1564–71.CrossRefGoogle Scholar
  29. 29.
    Wood KC, Chuang HF, Batten RD, Lynn DM, Hammond PT. (2006) Controlling interlayer diffusion to achieve sustained, multiagent delivery from layer-by-layer thin films. Proc. Natl. Acad. Sci. U. S. A. 103:10207–12.CrossRefGoogle Scholar
  30. 30.
    Blacklock J, You YZ, Zhou QH, Mao G, Oupicky D. (2009) Gene delivery in vitro and in vivo from bioreducible multilayered polyelectrolyte films of plasmid DNA. Biomaterials. 30:939–50.CrossRefGoogle Scholar
  31. 31.
    Jessel N, et al. (2006) Multiple and time-scheduled in situ DNA delivery mediated by beta-cyclodextrin embedded in a polyelectrolyte multilayer. Proc. Natl. Acad. Sci. U. S. A. 103:8618–21.CrossRefGoogle Scholar
  32. 32.
    Jewell CM, Zhang J, Fredin NJ, Lynn DM. (2005) Multilayered polyelectrolyte films promote the direct and localized delivery of DNA to cells. J. Control Release. 106:214–23.CrossRefGoogle Scholar
  33. 33.
    Meyer F, Ball V, Schaaf P, Voegel JC, Ogier J. (2006) Polyplex-embedding in polyelectrolyte multilayers for gene delivery. Biochim. Biophys. Acta. 1758:419–22.CrossRefGoogle Scholar
  34. 34.
    Zhang J, Chua LS, Lynn DM. (2004) Multilayered thin films that sustain the release of functional DNA under physiological conditions. Langmuir. 20:8015–21.CrossRefGoogle Scholar
  35. 35.
    Dimitrova M, et al. (2008) Sustained delivery of siRNAs targeting viral infection by cell-degradable multilayered polyelectrolyte films. Proc. Natl. Acad. Sci. U. S. A. 105:16320–5.CrossRefGoogle Scholar
  36. 36.
    Neumann B, et al. (2006) High-throughput RNAi screening by time-lapse imaging of live human cells. Nat. Methods. 3:385–90.CrossRefGoogle Scholar
  37. 37.
    Nolte A, et al. (2009) Optimized basic conditions are essential for successful siRNA transfection into primary endothelial cells. Oligonucleotides. 19:141–50.CrossRefGoogle Scholar
  38. 38.
    Marin V, Kaplanski G, Gres S, Farnarier C, Bongrand P. (2001) Endothelial cell culture: protocol to obtain and cultivate human umbilical endothelial cells. J. Immunol. Methods. 254:183–90.CrossRefGoogle Scholar
  39. 39.
    Relou IA, Damen CA, van der Schaft DW, Groenewegen G, Griffioen AW. (1998) Effect of culture conditions on endothelial cell growth and responsiveness. Tissue Cell. 30:525–30.CrossRefGoogle Scholar
  40. 40.
    Walker T, Wendel HP, Tetzloff L, Heidenreich O, Ziemer G. (2005) Suppression of ICAM-1 in human venous endothelial cells by small interfering RNAs. Eur. J. Cardiothorac. Surg. 28:816–20.CrossRefGoogle Scholar
  41. 41.
    Rozen S, Skaletsky H. (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol. Biol. 132:365–86.PubMedPubMedCentralGoogle Scholar
  42. 42.
    Ley K, Laudanna C, Cybulsky MI, Nourshargh S. (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat. Rev. Immunol. 7:678–89.CrossRefGoogle Scholar
  43. 43.
    Ley K, Miller YI, Hedrick CC. (2011) Monocyte and macrophage dynamics during atherogenesis. Arterioscler. Thromb. Vasc. Biol. 31:1506–16.CrossRefGoogle Scholar
  44. 44.
    Bigi A, Cojazzi G, Panzavolta S, Roveri N, Rubini K. (2002) Stabilization of gelatin films by crosslinking with genipin. Biomaterials. 23:4827–32.CrossRefGoogle Scholar
  45. 45.
    Walker T, et al. (2009) Graft protection in bypass surgery: siRNA-mediated silencing of adhesion molecules. Oligonucleotides. 19:15–21.CrossRefGoogle Scholar
  46. 46.
    Dykxhoorn DM, Novina CD, Sharp PA. (2003) Killing the messenger: short RNAs that silence gene expression. Nat. Rev. Mol. Cell. Biol. 4:457–67.CrossRefGoogle Scholar
  47. 47.
    Tepe G, et al. (2008) Local delivery of paclitaxel to inhibit restenosis during angioplasty of the leg. N. Engl. J. Med. 358:689–99.CrossRefGoogle Scholar
  48. 48.
    Werk M, et al. (2008) Inhibition of restenosis in femoropopliteal arteries: paclitaxel-coated versus uncoated balloon: femoral paclitaxel randomized pilot trial. Circulation. 118:1358–65.CrossRefGoogle Scholar
  49. 49.
    San Juan A, et al. (2009) Development of a functionalized polymer for stent coating in the arterial delivery of small interfering RNA. Biomacromolecules. 10:3074–80.CrossRefGoogle Scholar

Copyright information

© The Feinstein Institute for Medical Research 2011

Authors and Affiliations

  • Andrea Nolte
    • 1
  • Tobias Walker
    • 1
  • Martina Schneider
    • 1
  • Oya Kray
    • 1
  • Meltem Avci-Adali
    • 1
  • Gerhard Ziemer
    • 1
  • Hans Peter Wendel
    • 1
  1. 1.Department of Thoracic, Cardiac and Vascular SurgeryTuebingen University HospitalTuebingenGermany

Personalised recommendations