title: FGF-FGFR signaling mediated through glycosaminoglycans in microtiter plate and cell-based microarray platforms.
abstract: Fibroblast growth factor (FGF) signals cell growth through its interaction with a fibroblast growth factor receptor (FGFR) and a glycosaminoglycn (GAG) coreceptor. Here, we examine the signaling of five different FGFs (1, 2, 6, 8, and 8b) through FGFR3c. A small library of GAG and GAG-derivative coreceptors are screened to understand better the structure-activity relationship of these coreceptors on signaling. Initially, data were collected in a microtiter plate well-based cell proliferation assay. In an effort to reduce reagent requirements and improve assay throughput, a cell-based microarray platform was developed. In this cell-based microarray, FGFR3c-expressing cells were printed in alginate hydrogel droplets of ∼30 nL and incubated with FGF and GAG. Heparin was the most effective GAG coreceptor for all FGFs studied. Other GAGs, such as 2-O-desulfated heparin and chondroitin sulfate B, were also effective coreceptors. Signaling by FGF 8 and FGF 8b showed the widest tolerance for coreceptor structure. Finally, this on-chip cell-based microarray provides comparable data to a microtiter plate well-based assay, demonstrating that the coreceptor assay can be converted into a high-throughput assay.
authors.affiliation: Department of Chemical and Biological Engineering, ‡Department of Biomedical Engineering, §Department of Biology, llDepartment of Material Sciences, and ⊥Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute , Troy, New York 12180, United States.
authors.forename: Eric
authors.initials: E
authors.lastname: Sterner
authors.forename: Luciana
authors.initials: L
authors.lastname: Meli
authors.forename: Seok-Joon
authors.initials: SJ
authors.lastname: Kwon
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
mesh_terms: D000818:Animals; D002236:Carbohydrate Conformation; D049109:Cell Proliferation; D002478:Cells, Cultured; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D051379:Mice; D046228:Microarray Analysis; D008969:Molecular Sequence Data; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D006025:Glycosaminoglycans; D017468:Receptors, Fibroblast Growth Factor; D011994:Recombinant Proteins; D005346:Fibroblast Growth Factors; D006493:Heparin