Display options
Cite
Askou AL, Aagaard L, Kostic C, et al. Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors. Mol Ther Methods Clin Dev. 2015;2:14064doi: 10.1038/mtm.2014.64.
Askou, A. L., Aagaard, L., Kostic, C., Arsenijevic, Y., Hollensen, A. K., Bek, T., Jensen, T. G., Mikkelsen, J. G., & Corydon, T. J. (2015). Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors. Molecular therapy. Methods & clinical development, 214064. https://doi.org/10.1038/mtm.2014.64
Askou, Anne Louise, et al. "Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors." Molecular therapy. Methods & clinical development vol. 2 (2015): 14064. doi: https://doi.org/10.1038/mtm.2014.64
Askou AL, Aagaard L, Kostic C, Arsenijevic Y, Hollensen AK, Bek T, Jensen TG, Mikkelsen JG, Corydon TJ. Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors. Mol Ther Methods Clin Dev. 2015 Jan 28;2:14064. doi: 10.1038/mtm.2014.64. eCollection 2015. PMID: 26052532; PMCID: PMC4449022.
Copy Download .nbib Format: NLM
Share it on

Mol Ther Methods Clin Dev. 2015 Jan 28;2:14064. doi: 10.1038/mtm.2014.64. eCollection 2015.

Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors.

Molecular therapy. Methods & clinical development

Anne Louise Askou, Lars Aagaard, Corinne Kostic, Yvan Arsenijevic, Anne Kruse Hollensen, Toke Bek, Thomas Gryesten Jensen, Jacob Giehm Mikkelsen, Thomas Juhl Corydon

Affiliations

  1. Department of Biomedicine, Aarhus University , Aarhus, Denmark.
  2. Department of Ophthalmology, Unit of Gene Therapy and Stem Cell Biology, University of Lausanne, Jules-Gonin Eye Hospital , Lausanne, Switzerland.
  3. Department of Ophthalmology, Aarhus University Hospital , Aarhus, Denmark.

PMID: 26052532 PMCID: PMC4449022 DOI: 10.1038/mtm.2014.64

Abstract

Lentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters from a single vector have certain limitations that affect transgene expression levels and/or vector titers. In this study, we describe a novel vector design that facilitates combined expression of therapeutic RNA- and protein-based antiangiogenic factors as well as a fluorescent reporter from back-to-back RNApolII-driven expression cassettes. This configuration allows effective production of intron-embedded miRNAs that are released upon transduction of target cells. Exploiting such multigenic lentiviral vectors, we demonstrate robust miRNA-directed downregulation of vascular endothelial growth factor (VEGF) expression, leading to reduced angiogenesis, and parallel impairment of angiogenic pathways by codelivering the gene encoding pigment epithelium-derived factor (PEDF). Notably, subretinal injections of lentiviral vectors reveal efficient retinal pigment epithelium-specific gene expression driven by the VMD2 promoter, verifying that multigenic lentiviral vectors can be produced with high titers sufficient for in vivo applications. Altogether, our results suggest the potential applicability of combined miRNA- and protein-encoding lentiviral vectors in antiangiogenic gene therapy, including new combination therapies for amelioration of age-related macular degeneration.

References

  1. Mol Ther. 2008 Mar;16(3):557-64 - PubMed
  2. J Pharmacol Exp Ther. 2012 Jul;342(1):131-9 - PubMed
  3. Hum Gene Ther. 2006 Feb;17(2):167-76 - PubMed
  4. J Gene Med. 2012 Nov;14(11):632-41 - PubMed
  5. Retina. 2007 Feb;27(2):133-40 - PubMed
  6. Mol Ther. 2005 Oct;12(4):659-68 - PubMed
  7. Mol Ther. 2009 Oct;17(10):1743-53 - PubMed
  8. Ophthalmol Eye Dis. 2010 Dec 19;2:75-83 - PubMed
  9. Circ Res. 2012 Nov 9;111(11):1421-33 - PubMed
  10. Hum Gene Ther. 2012 Sep;23(9):980-91 - PubMed
  11. Retina. 2009 Jun;29(6 Suppl):S49-50 - PubMed
  12. Nucleic Acids Res. 2007;35(15):5154-64 - PubMed
  13. Mol Ther Nucleic Acids. 2012 Dec 18;1:e63 - PubMed
  14. Br J Ophthalmol. 1997 Feb;81(2):154-62 - PubMed
  15. Invest Ophthalmol Vis Sci. 2006 Aug;47(8):3496-504 - PubMed
  16. Hum Gene Ther. 2009 Jan;20(1):31-9 - PubMed
  17. Ophthalmology. 2012 Sep;119(9):1867-73 - PubMed
  18. Eye (Lond). 2012 Aug;26(8):1099-105 - PubMed
  19. Am J Ophthalmol. 2010 Jul;150(1):33-39.e2 - PubMed
  20. Biochem Biophys Res Commun. 2005 Jan 14;326(2):387-94 - PubMed
  21. N Engl J Med. 2008 May 22;358(21):2231-9 - PubMed
  22. Gene Ther. 2010 Mar;17(3):400-11 - PubMed
  23. Ophthalmology. 2013 Oct;120(10 ):2029-34 - PubMed
  24. J Gene Med. 2012 May;14(5):328-38 - PubMed
  25. Hum Gene Ther. 2008 Oct;19(10):979-90 - PubMed
  26. Mol Vis. 2003 May 30;9:210-6 - PubMed
  27. Gene Ther. 2008 Dec;15(23):1536-49 - PubMed
  28. Nat Med. 2006 Mar;12(3):348-53 - PubMed
  29. Biochem J. 1998 Apr 1;331 ( Pt 1):309-16 - PubMed
  30. Gene Ther. 2011 Jul;18(7):666-73 - PubMed
  31. J Biol Chem. 2010 Aug 27;285(35):26933-44 - PubMed
  32. Mol Genet Metab. 2005 Aug;85(4):260-70 - PubMed
  33. Nucleic Acids Res. 2014 Feb;42(4):e28 - PubMed
  34. Nature. 2008 Apr 3;452(7187):591-7 - PubMed
  35. Mol Ther. 2011 Feb;19(2):326-34 - PubMed
  36. J Med Genet. 2007 Aug;44(8):509-15 - PubMed
  37. N Engl J Med. 2008 May 22;358(21):2240-8 - PubMed
  38. J Biol Chem. 2004 Apr 30;279(18):19064-73 - PubMed
  39. Nature. 2006 May 25;441(7092):537-41 - PubMed
  40. Nat Cell Biol. 2008 Mar;10(3):329-37 - PubMed

Publication Types