title: Toxicity and anti-angiogenicity evaluation of Pak1 inhibitor IPA-3 using zebrafish embryo model.
abstract: p21-activated kinase 1 (Pak1)-a key node protein kinase regulating various cellular process including angiogenesis-has been recognised to be a therapeutic target for multitude of diseases, and hence, various small molecule inhibitors targeting its activity have been tested. However, the direct toxic and anti-angiogenic effects of these pharmacologic agents have not been examined. In this study, we evaluate the translational efficacy of Pak1 inhibitor IPA-3 using zebrafish toxicity model system to stratify its anti-angiogenic potential and off-target effects to streamline the compound for further therapeutic usage. The morphometric analysis has shown explicit delay in hatching, tail bending, pericardial sac oedema and abnormal angiogenesis. We provide novel evidence that Pak1 inhibitor could act as anti-angiogenic agents by impeding the development of sub-intestinal vessel (SIV) and intersegmental vessels (ISVs) by suppressing the expression of vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR2), neurophilin 1 (NRP1) and its downstream genes matrix metalloproteinase (MMP)-2 and MMP-9. Knockdown studies using 2-O-methylated oligoribonucleotides targeting Pak1 also revealed similar phenotypes with inhibition of angiogenesis accompanied with deregulation of major angiogenic factor and cardiac-specific genes. Taken together, our findings indicate that Pak1 signalling facilitates enhanced angiogenesis and also advocated the design and use of small molecule inhibitors of Pak1 as potent anti-angiogenic agents and suggest their utility in combinatorial therapeutic approaches targeting anomalous angiogenesis.
authors: Department of Genetics, Dr. ALM PGIBMS, University of Madras, Chennai, 600 113, India.
authors: Sankar
authors: S
authors: Jagadeeshan
authors: Department of Genetics, Dr. ALM PGIBMS, University of Madras, Chennai, 600 113, India.
authors: R Vivek
authors: RV
authors: Sagayaraj
authors: Department of Genetics, Dr. ALM PGIBMS, University of Madras, Chennai, 600 113, India.
authors: Navina
authors: N
authors: Paneerselvan
authors: Prism Foundation, Palissery P.O, Thrissur, 680 027, Kerala, India.
authors: Syed Shabin
authors: SS
authors: Ghouse
authors: Department of Genetics, Dr. ALM PGIBMS, University of Madras, Chennai, 600 113, India. r_malathi@hotmail.com.
authors: Raghunathan
authors: R
authors: Malathi
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D001808:Blood Vessels / Q000187:drug effects / Q000196:embryology; D004220:Disulfides / Q000737:chemistry / Q000633:toxicity*; D004625:Embryo, Nonmammalian / Q000187:drug effects* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D023421:Models, Animal; D009284:Naphthols / Q000737:chemistry / Q000633:toxicity*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018675:Toxicity Tests; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D015027:Zebrafish / Q000196:embryology*; D054462:p21-Activated Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D004220:Disulfides; C541795:IPA-3 compound; D009284:Naphthols; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D054462:p21-Activated Kinases
keywords: Anti-angiogenesis; IPA-3; Pak1; Toxicity; VEGF; Zebrafish embryos