Preferences help
enabled [disable] Abstract
Number of results
2003 | 50 | 3 | 875-882
Article title

Construction of a bicistronic proangiogenic expression vector and its application in experimental angiogenesis in vivo.

Title variants
Languages of publication
Manipulation of angiogenesis in vivo is an example of successful gene therapy strategies. Overexpression of angiogenic genes like VEGF, FGF or PDGF causes new vessel formation and improves the clinical state of patients. Gene therapy is a very promising procedure but requires large amounts of pharmaceutical-grade plasmid DNA. In this regard we have constructed a bicistronic plasmid DNA vector encoding two proangiogenic factors, VEGF165 and FGF-2. The construct (pVIF) contains the internal ribosome entry site (IRES) of the encephalomyocarditis virus (ECMV) which permits both genes to be translated from a single bicistronic mRNA. The IRES sequence allows for a high efficiency of gene expression in vivo. The pVIF vector was characterized in vitro and in vivo. In vivo angiogenesis studies showed that the bicistronic vector encoding two proangiogenic factors induces the formation of new vessels significantly more than pVEGF165 or pFGF-2 alone. In our opinion the combined proangiogenic approach with VEGF165 and FGF-2 is more powerful and efficient than single gene therapy. We also postulate that IRES sequence can serve as a useful device improving efficiency of gene therapy.
Physical description
  • Cancer Center, Department of Cell Biology, W. Roentgena 5, 02-781 Warszawa, Poland
  • Cancer Center, Department of Cell Biology, W. Roentgena 5, 02-781 Warszawa, Poland
  • Cancer Center, Department of Cell Biology, W. Roentgena 5, 02-781 Warszawa, Poland
  • Attal J, Theron MC, Houdebine LM. (1999) The optimal use of IRES (internal ribosome entry site) in expression vectors. Genet Anal.; 15: 161-5.
  • Battegay EJ. (1995) Angiogenesis: mechanistic insights, neovascular diseases, and therapeutic prospects. J Mol Med.; 73: 333-46.
  • de Felipe P. (2002) Polycistronic viral vectors. Curr Gene Ther.; 2: 355-78.
  • Dulak J, Józkowicz A, Guevara I, Dembińska-Kieć A. (1999) Gene transfer of vascular endothelial growth factor and endothelial nitric oxide syntase - implications for gene therapy in cardiovascular diseases. Pol J Pharmacol.; 51: 233-7.
  • Garcia-Martinez C, Opolon P, Trochon V, Chianale C, Musset K, Lu H, Abitbol M, Perricaudet M, Ragot T. (1999) Angiogenesis induced in muscle by a recombinant adenovirus expressing functional isoforms of basic fibroblast growth factor. Gene Ther.; 6: 1210-21.
  • Gurtu V, Yan G, Zhang G. (1996) IRES bicistronic expression vectors for efficient creation of stable mammalian cell lines. Biochem Biophys Res Commun.; 229: 295-8.
  • Harjai KJ, Chowdhury P, Grines CL. (2002) Therapeutic angiogenesis: a fantastic new adventure. J Interv Cardiol.; 15: 223-30.
  • Hayes AJ. (1999) Antivascular therapy: a new approach to cancer treatment. BMJ.; 318: 853-6.
  • Ibukiyama C. (1996) Angiogenesis. Angiogenic therapy using fibroblast growth factor and vascular endothelial growth factor for ischemic vascular lesions. Jap Heart J.; 37: 285-300.
  • Isner JM, Asahara T. (1998) Therapeutic angiogenesis. Front Biosc.; 3: 49-69.
  • Isner JM, Pieczek A, Schainfeld R, Blair R, Haley L, Ashara T, Rosenfield K, Razvi S, Walsh K, Symes JF. (1996a) Clinical evidence of angiogenesis after arterial gene transfer of phVEGF165 in patient with ischaemic limb. Lancet.; 348: 370-4.
  • Isner JM, Walsh K, Symes J, Pieczek A, Takeshita S, Lowry J, Rosenfield K, Weir L, Brogi E, Jurayj D. (1996b) Arterial gene transfer for therapeutic angiogenesis in patients with peripheral artery disease. Hum Gene Ther.; 7: 959-88.
  • Kapturczak M, Zolotukin S, Cross J, Pileggi A, Molano RD, Jorgensen M, Byrne B, Flotte TR, Ellis T, Inveradi L, Ricordi C, Nick H, Atkinson M, Agarwal A. (2002) Transduction of human and mouse pancreatic islet cells using a bicistronic recombinant adeno-associated viral vector. Mol Ther.; 5: 154-60.
  • Kołsut P, Małecki M, Teresińska A, Religa Z, Janik P. (2003) Gene therapy of coronary artery disease. Ann Diagnost Paediatr Pathol.; 7: 19-23.
  • Małecki M, Janik P. (1999) Naczyniowo-śródbłonkowy czynnik wzrostu (VEGF) i jego potencjalne aplikacje terapeutyczne w zmianach naczyniowych i nowotworach. Pol Przeg Chir.; 71: 1291-7 (in Polish).
  • Małecki M, Janik P. (2002) Terapia genowa - próby zastosowań klinicznych. Terapia angiogenna. Ordynator Leków.; 5: 8-12 (in Polish).
  • Małecki M, Przybyszewska M, Janik P. (2001) In vitro and in vivo study of the expression vector encoding vascular endothelial growth factor. Arch Immunol Ther Exp (Warsz).; 49: 243-6.
  • Martinez-Salas E. (1999) Internal ribosome entry site biology and its use in expression vectors. Curr Opin Biotechnol.; 10: 458-64.
  • Mizuguchi H, Xu Z, Ishii-Watabe A, Uchida E, Hayakawa T. (2000) IRES-dependent second gene expression is significantly lower than cap-dependent first gene expression in a bicistronic vector. Mol Ther.; 1: 376-82.
  • Przybyszewska M, Miłoszewska J, Janik P. (1998) Angiogenesis induced by urothelial cells (HCV-29) and their v-ras and v-raf transfectants. Cancer Lett.; 131: 157-61.
  • Sidky YA, Auerbach R. (1975) Lymphocyte-induced angiogenesis: a quantitative and sensitive assay of the graft-vs-host reaction. J Exp Med.; 141: 1084-100.
  • Singh G, Parker S, Hobart P. (2002) The development of a bicistronic plasmid DNA vaccine for B-cell lymphoma. Vaccine.; 20: 1400-11.
  • Szala S, Radzikowski C. (1997) Podloze molekularne angiogenezy nowotworow. Nowotwory.; 47: 1-19 (in Polish).
  • Vagner S, Galy B, Pyronnet S. (2001) Irresistible IRES. EMBO Rep.; 2: 893-8.
  • Wiznerowicz M, Fong AZ, Hawley RG, Mackiewicz A. (1998) Development of a double-copy bicistronic retroviral for human gene therapy. Adv Exp Med Biol.; 451: 441-7.
  • Zhang H, Harris AL. (1998) Anti-angiogenic therapies in cancer clinical trials. Exp Opin Invest Drugs.; 7: 1629-55.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.