Delivery of hypoxia-inducible VEGF gene to rat islets using polyethylenimine.
Islet transplantation is a promising strategy for treatment of diabetes. However, islets are exposed to hypoxia in the process of isolation and transplantation and prone to apoptosis. Vascular endothelial growth factor (VEGF) gene transfer is one of the promising strategies to address this problem. However, VEGF expression in the cells under normoxia is undesirable since it may induce pathological angiogenesis. Therefore, VEGF expression should be regulated to avoid this problem. In this study, hypoxia-inducible VEGF gene was transferred to islets using a non-viral carrier. Rat islets were transfected with high molecular weight PEI (25 kDa, PEI25K), low molecular weight PEI (2 kDa, PEI2K), and polyamidoamine dendrimer (PAMAM). PEI25K had higher transfection efficiency to rat islets than PAMAM or PEI2K. The hypoxia-inducible gene expression vector, pRTP801-Luc or pRTP801-VEGF was transferred to rat islets using PEI25K. Transfection assay with pRTP801-Luc showed that luciferase expression was induced in rat islets under hypoxia. In addition, transfer of pRTP801-VEGF showed that VEGF gene expression was higher under hypoxia than normoxia in rat islets. In conclusion, delivery of pRTP801-VEGF using PEI25K induces VEGF level specifically under hypoxia and may be useful for the development of anti-apoptotic strategies for islet transplantation.
Kim
Hyun Ah
HA
Department of Bioengineering, College of Engineering Hanyang University, Seoul, South Korea.
Lee
Byung-Wan
BW
Kang
Dongchul
D
Kim
Jae Hyeon
JH
Ihm
Sung Hee
SH
Lee
Minhyung
M
D000818:Animals; D018929:Cell Culture Techniques; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D050091:Dendrimers; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D000860:Hypoxia / Q000378:metabolism; D016381:Islets of Langerhans Transplantation / Q000473:pathology*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D011073:Polyamines / Q000737:chemistry / Q000378:metabolism; D011094:Polyethyleneimine / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D013552:Swine; D014157:Transcription Factors; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
D004268:DNA-Binding Proteins; C000632146:Ddit4 protein, rat; D050091:Dendrimers; C104700:PAMAM Starburst; D011073:Polyamines; D012097:Repressor Proteins; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; D011094:Polyethyleneimine; D008156:Luciferases