Intravenous injection of phagocytes transfected ex vivo with FGF4 DNA/biodegradable gelatin complex promotes angiogenesis in a rat myocardial ischemia/reperfusion injury model.
Conventional gene therapies still present difficulties due to poor tissue-targeting, invasiveness of delivery, method, or the use of viral vectors. To establish the feasibility of using non-virally ex vivo transfected phagocytes to promote angiogenesis in ischemic myocardium, gene-transfection into isolated phagocytes was performed by culture with positively charged gelatin impregnated with plasmid DNA. A high rate of gene transfection was achieved in rat macrophages and human monocytes, but not in mouse fibroblasts. The efficiency was 68 +/- 11% in rat macrophages and 78 +/- 8% in human monocytes. Intravenously injected phagocytes accumulated predominantly in ischemic tissue (13 +/- 8%) and spleen (84 +/- 6%), but negligibly in other organs in rodents. The efficiency of accumulation in the target ischemic tissue reached more than 86% on direct local tissue injection. In a rat model of myocardial ischemia-reperfusion, intravenous injection of fibroblast growth factor 4 (FGF4)-gene-transfected macrophages significantly increased regional blood flow in the ischemic myocardium (78 +/- 7.1 % in terms of flow ratio of ischemic/non-ischemic myocardium) compared with intravenous administration of saline (36 +/- 11%) or nontransfected macrophages (42 +/- 12 %), or intramuscular administration of naked DNA encoding FGF4 (75 +/- 18 %). Enhanced angiogenesis in the ischemic tissue we confirmed histologically. Similarly, intravenous injection of FGF4-gene-transfected monocytes enhanced regional blood flow in an ischemic hindlimb model in mice (93 +/- 22 %), being superior to the three other treatments described above (38 +/- 12, 39 +/- 15, and 55 +/- 12%, respectively). Phagocytes transfected ex vivo with FGF4 DNA/gelatin promoted angiogenesis. This approach might have potential for non-viral angiogenic gene therapy.
Fukuyama
Naoto
N
Depts of Physiology, Tokai University School of Medicine, Isehara, 259-1193, Japan. fukuyama@is.icc.u-tokai.ac.jp
Tanaka
Etsuro
E
Tabata
Yasuhiko
Y
Fujikura
Hisanori
H
Hagihara
Masao
M
Sakamoto
Hiromi
H
Ando
Kiyoshi
K
Nakazawa
Hiroe
H
Mori
Hidezo
H
D000818:Animals; D003326:Coronary Circulation / Q000187:drug effects / Q000235:genetics; D004195:Disease Models, Animal; D005240:Feasibility Studies; D051519:Fibroblast Growth Factor 4 / Q000494:pharmacology* / Q000627:therapeutic use; D005780:Gelatin / Q000008:administration & dosage; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007275:Injections, Intravenous; D008297:Male; D051379:Mice; D015428:Myocardial Reperfusion Injury / Q000628:therapy*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010586:Phagocytes; D051381:Rats; D011916:Rats, Inbred F344; D012039:Regional Blood Flow; D014162:Transfection
D051519:Fibroblast Growth Factor 4; D005780:Gelatin