Pharmacologically induced angiogenesis in transgenic zebrafish.
The rapid vascularisation of biomaterials and engineered tissue after implantation is a current unmet need. To this end, we explored the pharmacological option of inducing neovascularisation using compounds that inhibit hypoxia-induced factor-1alpha prolyl hydroxylase. This stabilises hypoxia inducible factor-1alpha and therefore de-repress the transcription of various angiogenic genes. In the quest for a small vertebrate model allowing for in vivo screening we exposed (TG(Fli1:EGFP)) transgenic zebrafish embryos exhibiting fluorescent blood vessels to hydralazine hydrochloride and 2,4-pyridine dicarboxylic acid from 6hpf to 72hpf by immersion. Live observation of embryos revealed that the substances induced formation of ectopic blood vessels in the subintestinal vessel basket. We confirmed the HIF-stabilising effects biochemically in human fibroblasts and with an in vitro angiogenesis fibroblast/HUVEC co-culture model. Cross-inhibition of collagen prolyl hydroxylase was confirmed by reduced collagen secretion by fibroblasts and reduced collagen content of zebrafish embryos.
Raghunath
Michael
M
Tissue Modulation Laboratory, NUS Tissue Engineering Programme, Life Science Institute, National University of Singapore, Singapore. bierm@nus.edu.sg
Sy Wong
Yuan
Y
Farooq
Muhammad
M
Ge
Ruowen
R
D043925:Angiogenesis Inducing Agents / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D001808:Blood Vessels / Q000254:growth & development; D002460:Cell Line; D003094:Collagen / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D006830:Hydralazine / Q000494:pharmacology*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D023421:Models, Animal; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011393:Procollagen-Proline Dioxygenase / Q000037:antagonists & inhibitors*; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics; D011725:Pyridines / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000502:physiology
D043925:Angiogenesis Inducing Agents; D004791:Enzyme Inhibitors; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D051789:Proto-Oncogene Protein c-fli-1; D011725:Pyridines; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006830:Hydralazine; C055293:2,4-pyridinedicarboxylic acid; D003094:Collagen; D011393:Procollagen-Proline Dioxygenase