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Chemical Genetic Analysis of Glycome Regulation of Vasculogenesis
Author(s) -
Piecewicz Stephanie Marie,
Hentschel Dirk,
Sengupta Shiladitya
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1163
Subject(s) - vasculogenesis , pericyte , microbiology and biotechnology , endothelial stem cell , angiogenesis , biology , embryoid body , zebrafish , matrigel , glycome , embryonic stem cell , extracellular matrix , neovascularization , chemistry , stem cell , progenitor cell , biochemistry , in vitro , cancer research , adult stem cell , glycan , glycoprotein , gene
Limited knowledge exists on the role of the extracellular ‘glyco’ matrix on neovascularization. In this study, we integrated our novel glycome‐sequencing platform with a chemical‐genetic approach to study the role of cell‐surface heparan sulfate glycosaminoglycans (HSGAGs) in vasculogenesis in vitro and in vivo. An embryonic stem cell‐embryoid body was monitored using stemness marker Oct‐4 or endothelial cell markers VEGFR2, vWF, VE‐cadherin and eNOS. RT‐PCR and immunolabeling revealed a progressive differentiation (P<0.01 vs Day 0) into endothelial cells. Compositional analysis revealed an amplification of cell surface HSGAGs matched by an amplification of HSGAG synthetic enzyme, N‐deacytylase/N‐sulfotransferase1 transcripts during differentiation. Inhibitions of sulfation or enzymatic‐disruptions of HSGAGs inhibited the differentiation into endothelial cells, as quantified by decreased levels of endothelial cell markers (P<0.01 vs untreated controls). This was recapitulated in vivo , where morpholino‐injected zebrafish embryos exhibited altered vessel anatomy, hemorrhage in the caudal vein plexus, and lack of lumen formation of intersegmental vessels, which was recovered by the addition of exogenous HSGAGs. Our study demonstrates glycome regulation of vasculogenesis, opening up the possibility of regulating neovascularization through engineered cellular microenvironment.