title: Novel anti IGFBP2 single chain variable fragment inhibits glioma cell migration and invasion.
abstract: Insulin like growth factor binding protein 2 (IGFBP2) is highly up regulated in glioblastoma (GBM) tissues and has been one of the prognostic indicators. There are compelling evidences suggesting important roles for IGFBP2 in glioma cell proliferation, migration and invasion. Extracellular IGFBP2 through its carboxy terminal arginine glycine aspartate (RGD) motif can bind to cell surface α5β1 integrins and activate pathways downstream to integrin signaling. This IGFBP2 activated integrin signaling is known to play a crucial role in IGFBP2 mediated invasion of glioma cells. Hence a molecular inhibitor of carboxy terminal domain of IGFBP2 which can inhibit IGFBP2-cell surface interaction is of great therapeutic importance. In an attempt to develop molecular inhibitors of IGFBP2, we screened single chain variable fragment (scFv) phage display libraries, Tomlinson I (Library size 1.47 × 10(8)) and Tomlinson J (Library size 1.37 × 10(8)) using human recombinant IGFBP2. After screening we obtained three IGFBP2 specific binders out of which one scFv B7J showed better binding to IGFBP2 at its carboxy terminal domain, blocked IGFBP2-cell surface association, reduced activity of matrix metalloprotease 2 in the conditioned medium of glioma cells and inhibited IGFBP2 induced migration and invasion of glioma cells. We demonstrate for the first time that in vitro inhibition of extracellular IGFBP2 activity by using human scFv results in significant reduction of glioma cell migration and invasion. Therefore, the inhibition of IGFBP2 can serve as a potential therapeutic strategy in the management of GBM.
authors.affiliation: Department of Molecular Reproduction Development and Genetics, Indian Institute of Science, Bangalore, 560012, India.
authors.forename: Shilpa S
authors.initials: SS
authors.lastname: Patil
authors.forename: Reema
authors.initials: R
authors.lastname: Railkar
authors.forename: Monalisa
authors.initials: M
authors.lastname: Swain
authors.forename: Hanudatta S
authors.initials: HS
authors.lastname: Atreya
authors.forename: Rajan R
authors.initials: RR
authors.lastname: Dighe
authors.forename: Paturu
authors.initials: P
authors.lastname: Kondaiah
mesh_terms: D017209:Apoptosis; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D002448:Cell Adhesion; D002465:Cell Movement; D049109:Cell Proliferation; D004797:Enzyme-Linked Immunosorbent Assay; D015972:Gene Expression Regulation, Neoplastic; D005910:Glioma / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D006801:Humans; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000037:antagonists & inhibitors* / Q000276:immunology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D009361:Neoplasm Invasiveness; D019151:Peptide Library; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D057127:Single-Chain Antibodies / Q000276:immunology / Q000494:pharmacology*; D020349:Surface Plasmon Resonance; D014407:Tumor Cells, Cultured
chemical_list: D018971:Insulin-Like Growth Factor Binding Protein 2; D019151:Peptide Library; D012333:RNA, Messenger; D057127:Single-Chain Antibodies; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2