In vivo molecular validation of VEGF inhibitor extracted from marine Actinomycetes in zebrafish embryos

Abstract

Angiogenesis, the formation of new blood vessels by sprouting from pre-existing endothelium, is a significant component in tumor progression. Marine natural products have been proved to be a rich source of novel compounds in drug discovery especially in the area of small molecule based targeted anticancer chemotherapeutics identification. Symbiotic actinomycetes from marine tunicates are having unique environmental niche that arise as a new and valuable source of inhibitors of angiogenesis. In the present study, actinomycetes strains were isolated from the areas of Muttom, Kanyakumari, Tamil Nadu, India and it was screened for the production of bioactive compounds. The bioactive compounds obtained were treated on zebrafish embryos at 50 % epiboly stage and maintained for 72 hours post fertilization (hpf). RBC staining was done at 72 hpf to study the formation of blood vessel in embryos treated with biomolecules and RT-PCR was carried out to study mRNA expression of vascular endothelial growth factor (<em>vegfaa</em>) at 72 hpf. Biomolecules (T1 and T2) extracted from marine actinomyetes showed anti-angiogenic property by reducing the Intersegmental vessels (ISVs) (main angiogenic blood vessel) at 5 µM concentration. RT-PCR study reveals the reduction in mRNA expression of <em>vegfaa</em>. Our study emphasizes the importance of potential of anti-angiogenic compounds from marine tunicate symbiotic actinomycetes using zebrafish as <em>in vivo</em> model system.