IRES-based vector coexpressing FGF2 and Cyr61 provides synergistic and safe therapeutics of lower limb ischemia.
Due to the lack of an adequate conventional therapy against lower limb ischemia, gene transfer for therapeutic angiogenesis is seen as an attractive alternative. However, the possibility of side effects, due to the expression of large amounts of angiogenic factors, justifies the design of devices that express synergistic molecules in low controlled doses. We have developed an internal ribosome entry site (IRES)-based bicistronic vector expressing two angiogenic molecules, fibroblast growth factor 2 (FGF2), and Cyr61. Through electrotransfer into the ApoE(-/-) mice hindlimb ischemic muscle model, we show that the IRES-based vector gives more stable expression than either monocistronic plasmid. Furthermore, laser Doppler analysis, arteriography, and immunochemistry clearly show that the bicistronic vector promotes a more abundant and functional revascularization than the monocistronic vectors, despite the fact that the bicistronic system produces 5-10 times less of each angiogenic molecule. Furthermore, although the monocistronic Cyr61 vector accelerates B16 melanoma growth in mice, the bicistronic vector is devoid of such side effects. Our results show an active cooperation of FGF2 and Cyr61 in therapeutic angiogenesis of hindlimb ischemia, and validate the use of IRES-based bicistronic vectors for the coexpression of controlled low doses of therapeutic molecules, providing perspectives for a safer gene therapy of lower limb ischemia.
Rayssac
Audrey
A
Inserm, U858, Toulouse, France.
Neveu
Charles
C
Pucelle
Mélanie
M
Van den Berghe
Loïc
L
Prado-Lourenco
Leonel
L
Arnal
Jean-François
JF
Chaufour
Xavier
X
Prats
Anne-Catherine
AC
D000818:Animals; D001057:Apolipoproteins E / Q000502:physiology; D055515:Cysteine-Rich Protein 61 / Q000235:genetics*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006614:Hindlimb / Q000098:blood supply*; D007511:Ischemia / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism; D018919:Neovascularization, Physiologic; D012270:Ribosomes / Q000235:genetics
D001057:Apolipoproteins E; C491850:CCN1 protein, mouse; D055515:Cysteine-Rich Protein 61; D016222:Fibroblast Growth Factor 2