title: An iminosugar-based heparanase inhibitor heparastatin (SF4) suppresses infiltration of neutrophils and monocytes into inflamed dorsal air pouches.
abstract: Local infiltration of inflammatory cells is regulated by a number of biological steps during which the cells likely penetrate through subendothelial basement membranes that contain heparan sulfate proteoglycans. In the present study, we examined whether administration of heparastatin (SF4), an iminosugar-based inhibitor of heparanase, could suppress local inflammation and degradation of heparan sulfate proteoglycans in basement membranes. In a carrageenan- or formyl peptide-induced dorsal air pouch inflammation model, the number of infiltrated neutrophils and monocytes was significantly lower in mice after topical administration of heparastatin (SF4). The concentration of chemokines MIP-2 and KC in pouch exudates of drug-treated mice was similar to control. In a zymosan-induced peritonitis model, the number of infiltrated cells was not altered in drug-treated mice. To further test how heparastatin (SF4) influences transmigration of inflammatory neutrophils, its suppressive effect on migration and matrix degradation was examined in vitro. In the presence of heparastatin (SF4), the number of neutrophils that infiltrated across a Matrigel-coated polycarbonate membrane was significantly lower, while the number of neutrophils passing through an uncoated membrane was not altered. Lysate of bone marrow-derived neutrophils released sulfate-radiolabeled macromolecules from basement membrane-like extracellular matrix, which was suppressed by heparastatin (SF4). Heparan sulfate degradation activity was almost completely abolished after incubation of lysate with protein G-conjugated anti-heparanase monoclonal antibody, strongly suggesting that the activity was due to heparanase-mediated degradation. Taken together, in a dorsal air pouch inflammation model heparastatin (SF4) potentially suppresses extravasation of inflammatory cells by impairing the degradation of basement membrane heparan sulfate.
authors.affiliation: Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Sue
authors.affiliation: Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan; One-stop Sharing Facility Center for Future Drug Discoveries, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan. Electronic address: higashi@mol.f.u-tokyo.ac.jp.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Higashi
authors.affiliation: Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shida
authors.affiliation: Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kogane
authors.affiliation: Institute of Microbial Chemistry (BIKAKEN), Kamiosaki 3-14-23, Shinagawa-ku, Tokyo 141-0021, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Nishimura
authors.affiliation: Institute of Microbial Chemistry (BIKAKEN), Kamiosaki 3-14-23, Shinagawa-ku, Tokyo 141-0021, Japan.
authors.forename: Hayamitsu
authors.initials: H
authors.lastname: Adachi
authors.affiliation: Institute of Zoology, Jagiellonian University, ul. Gronostajowa 9, 30-387 Krakow, Poland.
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Kolaczkowska
authors.affiliation: Institute of Zoology, Jagiellonian University, ul. Gronostajowa 9, 30-387 Krakow, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kepka
authors.affiliation: SBI Pharmaceuticals Co., Ltd., 1-6-1, Roppongi, Minato-ku, Tokyo 106-6019, Japan.
authors.forename: Motowo
authors.initials: M
authors.lastname: Nakajima
authors.affiliation: Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Biochemistry, Juntendo University School of Medicine, 2-1-1, Hongo, Bunkyo-ku, Tokyo 104-8560, Japan; Department of Breast and Endocrine Surgery, Juntendo University School of Medicine, 2-1-1, Hongo, Bunkyo-ku, Tokyo 104-8560, Japan. Electronic address: t-irimura@juntendo.ac.jp.
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Irimura
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000187:drug effects*; D002351:Carrageenan / Q000276:immunology; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000627:therapeutic use*; D005966:Glucuronidase / Q000037:antagonists & inhibitors*; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D050111:Imino Sugars / Q000138:chemical synthesis / Q000627:therapeutic use*; D007249:Inflammation / Q000188:drug therapy* / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D009000:Monocytes / Q000187:drug effects* / Q000502:physiology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000276:immunology; D009504:Neutrophils / Q000187:drug effects* / Q000502:physiology; D009557:Nipecotic Acids / Q000138:chemical synthesis / Q000627:therapeutic use*; D000099175:Heparanase
chemical_list: D002351:Carrageenan; D004791:Enzyme Inhibitors; D005966:Glucuronidase; D006497:Heparan Sulfate; D050111:Imino Sugars; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009557:Nipecotic Acids; D000099175:Heparanase; C099337:6-(trifluoroacetamido)-4,5-dihydroxypiperidine-3-carboxylic acid
keywords: Chemokines; Extracellular matrix; Heparanase; Heparastatin (SF4)