Powerful and controllable angiogenesis by using gene-modified cells expressing human hepatocyte growth factor and thymidine kinase.
OBJECTIVE
This study investigated the possibility of achieving angiogenesis by using gene-modified cells as a vector.


BACKGROUND
Although gene therapy for peripheral circulation disorders has been studied intensively, the plasmid or viral vectors have been associated with several disadvantages, including unreliable transfection and uncontrollable gene expression.


METHODS
Human hepatocyte growth factor (hHGF) and thymidine kinase (TK) expression plasmids were serially transfected into NIH3T3 cells, and permanent transfectants were selected (NIH3T3 + hHGF + TK). Unilateral hindlimb ischemia was surgically induced in BALB/c nude mice, and cells were transplanted into the thigh muscles. All effects were assessed at four weeks.


RESULTS
The messenger ribonucleic acid expression and protein production of hHGF were confirmed. Assay of growth inhibition by ganciclovir revealed that the 50% (median) inhibitory concentration of NIH3T3 + hHGF + TK was 1000 times lower than that of NIH3T3 + hHGF. The NIH3T3 + hHGF + TK group had a higher laser Doppler blood perfusion index, higher microvessel density, wider microvessel diameter, and lower rate of hindlimb necrosis, as compared with the plasmid- and adenovirus-mediated hHGF transfection groups or the NIH3T3 group. The newly developed microvessels were accompanied by smooth muscle cells, as well as endothelial cells, indicating that they were on the arteriolar or venular level. Laser Doppler monitoring showed that the rate of blood perfusion could be controlled by oral administration of ganciclovir. The transplanted cells completely disappeared in response to ganciclovir administration for four weeks.


CONCLUSIONS
Gene-modified cell transplantation therapy induced strong angiogenesis and collateral vessel formation that could be controlled externally with ganciclovir.
Hisaka
Yasuyo
Y
Institute for Advanced Cardiac Therapeutics, Tokyo, Japan.
Ieda
Masaki
M
Nakamura
Toshikazu
T
Kosai
Ken-ichiro
K
Ogawa
Satoshi
S
Fukuda
Keiichi
K
D016475:3T3 Cells; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003097:Collateral Circulation / Q000187:drug effects / Q000235:genetics; D004798:Enzymes / Q000008:administration & dosage; D015774:Ganciclovir / Q000494:pharmacology; D015316:Genetic Therapy / Q000379:methods*; D006133:Growth Substances / Q000008:administration & dosage*; D017228:Hepatocyte Growth Factor / Q000008:administration & dosage*; D006614:Hindlimb / Q000098:blood supply*; D006801:Humans; D007511:Ischemia / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000235:genetics; D013937:Thymidine Kinase / Q000008:administration & dosage
D000998:Antiviral Agents; D004798:Enzymes; D006133:Growth Substances; D017228:Hepatocyte Growth Factor; D013937:Thymidine Kinase; D015774:Ganciclovir