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Molecular Interaction of Neuropilin‐1 and 3‐O‐sulfated Heparan Sulfate Modulates Angiogenesis‐dependent Tumor Growth
Author(s) -
Painter Chelsea D.,
Clausen Thomas M.,
Park Jeeyoung,
Vasquez Gabriel J.,
Liu Jian,
Gordts Philip L.S.M.,
Esko Jeffrey D.
Publication year - 2020
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.2020.34.s1.07258
Subject(s) - heparan sulfate , neuropilin 1 , angiogenesis , perlecan , chemistry , sulfation , microbiology and biotechnology , biochemistry , fibroblast growth factor , syndecan 1 , glycosaminoglycan , receptor , biology , vascular endothelial growth factor , cancer research , cell , vegf receptors
Neuropilin‐1 (Nrp1) acts as a coreceptor with Vascular Endothelial Growth Factor Receptor (VEGFR) to facilitate binding of its ligand, Vascular Endothelial Growth Factor (VEGF). Heparan sulfate (HS) has been shown to be a cofactor facilitating the interaction of Nrp1 with VEGFR/VEGFs, but the functional significance of this interaction has not been established. Previous studies have shown that Nrp1 binding to heparan sulfate is enhanced by 3 ‐O‐ sulfation, a rare modification in which a sulfate group is installed at the C3 position of a glucosamine residue. Nrp1 mediates angiogenesis including branching and organization of vessels through its role in tip cell function and interactions with VEGFR/VEGFs. We hypothesize that heparan sulfate, particularly 3 ‐O ‐sulfation, is critical for Nrp1 function in developmental and pathological angiogenesis. We have created a series of Nrp1 mutants to identify the specific amino acids that interact with heparan sulfate, particularly those that interact with the 3 ‐O‐ sulfate group. Specific mutations in the b1b2 domains of Nrp1 decrease binding to heparin and cell surface heparan sulfate. These mutants also prevent heparin‐induced stabilization of the protein during thermal denaturation. Knock‐in mice expressing Neuropilin‐1 deficient in heparan sulfate binding (Nrp1 HSBD ) were created using CRISPR/Cas9 gene targeting. These mice are currently being evaluated for overall gross phenotype as well as proper branching and organization of developing vessels. Analysis of subcutaneous tumor formation revealed that Nrp1 HSBD mice exhibit a reduction in tumor growth compared to wildtype controls. This finding suggests that the interaction of Neuropilin‐1 with heparan sulfate modulates its function in pathological angiogenesis.