Reducible poly(amido ethylenediamine) for hypoxia-inducible VEGF delivery.
Delivery of the hypoxia-inducible vascular endothelial growth factor (RTP-VEGF) plasmid using a novel reducible disulfide poly(amido ethylenediamine) (SS-PAED) polymer carrier was studied in vitro and in vivo. In vitro transfection of primary rat cardiomyoblasts (H9C2) showed SS-PAED at a weighted ratio of 12:1 (polymer/DNA) mediates 16 fold higher expression of luciferase compared to an optimized bPEI control. FACS analysis revealed up to 57+/-2% GFP positive H9C2s. The efficiency of plasmid delivery to H9C2 using SS-PAED was found to depend upon glutathione (GSH) levels inside the cell. SS-PAED mediated delivery of RTP-VEGF plasmid produced significantly higher levels of VEGF expression (up to 76 fold) under hypoxic conditions compared to normoxic conditions in both H9C2 and rat aortic smooth muscle cells (A7R5). Using SS-PAED, delivery of RTP-VEGF was investigated in a rabbit myocardial infarct model using 100 mug RTP-VEGF. Results showed up to 4 fold increase in VEGF protein expression in the region of the infarct compared to injections of SS-PAED/RTP-Luc. In conclusion, SS-PAED mediated therapeutic delivery improves the efficacy of ischemia-inducible VEGF gene therapy both in vitro and in vivo and therefore, has potential for the promotion of neo-vascular formation and improvement of tissue function in ischemic myocardium.
Christensen
Lane V
LV
Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT 84112-5820, USA.
Chang
Chien-Wen
CW
Yockman
James W
JW
Conners
Rafe
R
Jackson
Heidi
H
Zhong
Zhiyuan
Z
Feijen
Jan
J
Bull
David A
DA
Kim
Sung Wan
SW
D000818:Animals; D015687:Cell Hypoxia; D002469:Cell Separation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005978:Glutathione / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008156:Luciferases; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D009203:Myocardial Infarction / Q000235:genetics / Q000378:metabolism; D017202:Myocardial Ischemia / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism; D010084:Oxidation-Reduction; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D011073:Polyamines / Q000737:chemistry / Q000378:metabolism*; D011094:Polyethyleneimine / Q000737:chemistry; D011817:Rabbits; D051381:Rats; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
D004220:Disulfides; D011073:Polyamines; D042461:Vascular Endothelial Growth Factor A; C520331:disulfide poly(amido ethylenediamine); D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D008156:Luciferases; D005978:Glutathione