Improved angiogenic response in pig heart following ischaemic injury using human skeletal myoblast simultaneously expressing VEGF165 and angiopoietin-1.
OBJECTIVE
To achieve angiogenic interaction between VEGF(165) and angiopoietin-1 (Ang-1) using a novel adenoviral bicistronic vector (Ad-Bic) encoding the two factors and delivered ex vivo using sex-mismatched human skeletal myoblasts.


RESULTS
A myocardial infarction model was developed in 29 female pigs; randomised into four groups: DMEM (group-1, n=6); Adenovirus null (Ad-null) vector-myoblast (group-2, n=5); Ad-Ang-1 myoblast (group 3, n=7) and Ad-Bic-myoblast (group-4, n=11). Three weeks later, 5 ml DMEM without myoblasts or containing 3 x 10(8) myoblasts carrying lac-z gene and transduced with Ad-null, Ad-Ang-1 or Ad-Bic were injected intra-myocardially in and around the infarct. 2D-echocardiography and fluorescent microsphere studies 6- and 12-weeks post-treatment revealed significantly improved cardiac performance and regional blood flow in groups 3 and 4. Histological studies and Y-chromosome analysis revealed extensive survival of lac-z positive myoblasts staining positive for human proteins in the pig heart. ELISA, immunostaining and RT-PCR revealed that Ad-Bic transduced myoblasts concomitantly but transiently expressed hVEGF(165) and Ang-1 both in vitro and in vivo. Double fluorescent immunostaining of the tissue sections for vWFactor-III and smooth muscle actin showed significantly higher vascular density of mature blood vessels per low power microscopic field in groups 3 and 4 at 6- and 12-weeks.


CONCLUSIONS
Our combined approach led to enhanced angiogenesis with a greater percentage of functionally mature blood vessels in a porcine heart.
Ye
Lei
L
Department of Surgery, National University of Singapore-119074, Singapore.
Haider
Husnain Kh
HKh
Jiang
Shujia
S
Tan
Ru San
RS
Ge
Ruowen
R
Law
Peter K
PK
Sim
Eugene K W
EK
D000256:Adenoviridae; D042683:Angiopoietin-1 / Q000235:genetics*; D000818:Animals; D001706:Biopsy; D001783:Blood Flow Velocity; D003326:Coronary Circulation; D017931:DNA Primers; D004195:Disease Models, Animal; D004562:Electrocardiography; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology*; D032446:Myoblasts / Q000502:physiology* / Q000637:transplantation*; D009203:Myocardial Infarction / Q000628:therapy*; D017202:Myocardial Ischemia / Q000503:physiopathology*; D018919:Neovascularization, Physiologic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
C467630:ANGPT1 protein, human; D042683:Angiopoietin-1; D017931:DNA Primers; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A