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Tuttolomondo M, Casella C, Hansen PL, et al. Human DMBT1-Derived Cell-Penetrating Peptides for Intracellular siRNA Delivery. Mol Ther Nucleic Acids. 2017;8:264-276doi: 10.1016/j.omtn.2017.06.020.
Tuttolomondo, M., Casella, C., Hansen, P. L., Polo, E., Herda, L. M., Dawson, K. A., Ditzel, H. J., & Mollenhauer, J. (2017). Human DMBT1-Derived Cell-Penetrating Peptides for Intracellular siRNA Delivery. Molecular therapy. Nucleic acids, 8264-276. https://doi.org/10.1016/j.omtn.2017.06.020
Tuttolomondo, Martina, et al. "Human DMBT1-Derived Cell-Penetrating Peptides for Intracellular siRNA Delivery." Molecular therapy. Nucleic acids vol. 8 (2017): 264-276. doi: https://doi.org/10.1016/j.omtn.2017.06.020
Tuttolomondo M, Casella C, Hansen PL, Polo E, Herda LM, Dawson KA, Ditzel HJ, Mollenhauer J. Human DMBT1-Derived Cell-Penetrating Peptides for Intracellular siRNA Delivery. Mol Ther Nucleic Acids. 2017 Sep 15;8:264-276. doi: 10.1016/j.omtn.2017.06.020. Epub 2017 Jun 29. PMID: 28918028; PMCID: PMC5514624.
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Mol Ther Nucleic Acids. 2017 Sep 15;8:264-276. doi: 10.1016/j.omtn.2017.06.020. Epub 2017 Jun 29.

Human DMBT1-Derived Cell-Penetrating Peptides for Intracellular siRNA Delivery.

Molecular therapy. Nucleic acids

Martina Tuttolomondo, Cinzia Casella, Pernille Lund Hansen, Ester Polo, Luciana M Herda, Kenneth A Dawson, Henrik J Ditzel, Jan Mollenhauer

Affiliations

  1. Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, 5000 Odense C, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark. Electronic address: [email protected].
  2. Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, 5000 Odense C, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark.
  3. Centre for BioNano Interactions, School of Chemistry, University College Dublin, Belfield, Dublin 4, Ireland.
  4. Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, 5000 Odense C, Denmark; Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark; Department of Oncology, Odense University Hospital, 5000 Odense C, Denmark. Electronic address: [email protected].

PMID: 28918028 PMCID: PMC5514624 DOI: 10.1016/j.omtn.2017.06.020

Abstract

Small interfering RNA (siRNA) is a promising molecule for gene therapy, but its therapeutic administration remains problematic. Among the recently proposed vectors, cell-penetrating peptides show great promise in in vivo trials for siRNA delivery. Human protein DMBT1 (deleted in malignant brain tumor 1) is a pattern recognition molecule that interacts with polyanions and recognizes and aggregates bacteria. Taking advantage of these properties, we investigated whether specific synthetic DMBT1-derived peptides could be used to formulate nanoparticles for siRNA administration. Using an electrophoretic mobility shift assay and UV spectra, we identified two DMBT1 peptides that could encapsulate the siRNA with a self- and co-assembly mechanism. The complexes were stable for at least 2 hr in the presence of either fetal bovine serum (FBS) or RNase A, with peptide-dependent time span protection. ζ-potential, circular dichroism, dynamic light scattering, and transmission electron microscopy revealed negatively charged nanoparticles with an average diameter of 10-800 nm, depending on the reaction conditions, and a spherical or rice-shaped morphology, depending on the peptide and β-helix conformation. We successfully transfected human MCF7 cells with fluorescein isothiocyanate (FITC)-DMBT1-peptide-Cy3-siRNA complexes. Finally, DMBT1 peptides encapsulating an siRNA targeting a fluorescent reporter gene showed efficient gene silencing in MCF7-recombinant cells. These results lay the foundation for a new research line to exploit DMBT1-peptide nanocomplexes for therapeutic siRNA delivery.

Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Keywords: DMBT1; RNAi; cell-penetrating peptides; cellular uptake; gene knockdown; nanoparticles; nanotherapy; siRNA delivery; silencing; transfection

References

  1. Mol Immunol. 1997 Feb;34(2):185-94 - PubMed
  2. Biophys J. 2010 Jul 21;99(2):619-28 - PubMed
  3. Adv Drug Deliv Rev. 2009 Aug 10;61(10):850-62 - PubMed
  4. Mol Ther. 2012 Mar;20(3):525-33 - PubMed
  5. Pharm Res. 2015 Apr;32(4):1462-74 - PubMed
  6. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2308-10 - PubMed
  7. J Biol Chem. 2004 Nov 12;279(46):47699-703 - PubMed
  8. Adv Drug Deliv Rev. 2007 Mar 30;59(2-3):75-86 - PubMed
  9. Eur J Immunol. 2009 Mar;39(3):833-42 - PubMed
  10. Oncogene. 1999 Nov 4;18(46):6233-40 - PubMed
  11. ACS Nano. 2013 May 28;7(5):3797-807 - PubMed
  12. Biochemistry. 1969 Oct;8(10):4108-16 - PubMed
  13. Biochem Pharmacol. 2006 Feb 28;71(5):702-10 - PubMed
  14. Curr Drug Deliv. 2017;14(1):36-46 - PubMed
  15. Chem Commun (Camb). 2011 Jan 7;47(1):188-90 - PubMed
  16. J Mol Med (Berl). 2004 Feb;82(2):144-52 - PubMed
  17. Mol Biosyst. 2007 Jan;3(1):43-50 - PubMed
  18. Methods Mol Biol. 2016;1364:291-310 - PubMed
  19. Nature. 2000 Jul 20;406(6793):322-4 - PubMed
  20. J Comput Chem. 2004 Oct;25(13):1605-12 - PubMed
  21. Mol Ther. 2009 Jan;17(1):95-103 - PubMed
  22. Electrophoresis. 1997 Dec;18(15):2714-23 - PubMed
  23. J Biol Chem. 2002 Aug 30;277(35):32109-15 - PubMed
  24. ACS Nano. 2014 Mar 25;8(3):1972-94 - PubMed
  25. J Struct Biol. 2001 May-Jun;134(2-3):117-31 - PubMed
  26. Peptides. 2014 Jul;57:78-94 - PubMed
  27. Nat Rev Drug Discov. 2009 Feb;8(2):129-38 - PubMed
  28. Cancer Res. 2000 Mar 15;60(6):1704-10 - PubMed
  29. J Control Release. 2010 Aug 3;145(3):272-80 - PubMed
  30. J Struct Biol. 1998;122(1-2):223-35 - PubMed
  31. J Biol Chem. 1997 May 23;272(21):13743-9 - PubMed
  32. Genes Chromosomes Cancer. 2002 Nov;35(3):242-55 - PubMed
  33. AAPS J. 2010 Dec;12(4):492-503 - PubMed
  34. Biotechnol Prog. 2008 Jul-Aug;24(4):957-63 - PubMed
  35. Biophys J. 2013 Apr 2;104(7):1595-604 - PubMed
  36. Nanoscale Res Lett. 2012 Jul 01;7(1):358 - PubMed
  37. Mol Biosyst. 2013 May;9(5):855-61 - PubMed
  38. Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):572-9 - PubMed
  39. Langmuir. 2012 Dec 21;28(51):17666-71 - PubMed
  40. Expert Opin Drug Deliv. 2016;13(3):373-87 - PubMed
  41. Expert Opin Drug Deliv. 2014 Sep;11(9):1481-95 - PubMed
  42. Biomed Res Int. 2013;2013:782041 - PubMed
  43. Arterioscler Thromb Vasc Biol. 2012 Feb;32(2):442-8 - PubMed
  44. Nat Biotechnol. 2014 Dec;32(12):1197-8 - PubMed
  45. Nat Protoc. 2006;1(6):2876-90 - PubMed
  46. Mol Ther. 2011 Feb;19(2):372-80 - PubMed
  47. Nat Mater. 2013 Nov;12(11):967-77 - PubMed
  48. J Control Release. 2014 Jan 28;174:126-36 - PubMed
  49. Molecules. 2015 Aug 03;20(8):14033-50 - PubMed
  50. Proc Natl Acad Sci U S A. 1999 Sep 14;96(19):10794-9 - PubMed
  51. J Biol Chem. 2000 Dec 22;275(51):39860-6 - PubMed
  52. Nat Genet. 1997 Sep;17(1):32-9 - PubMed
  53. PLoS One. 2013;8(2):e57601 - PubMed
  54. Int J Mol Sci. 2010;11(12):5212-33 - PubMed

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