title: Treatment of dermal fibroblasts with GPI-anchored human TIMP-1 protein moderates processes linked to scar formation.
abstract: Tissue inhibitors of metalloproteinases exhibit diverse physiological/biological functions including moderation of the proteolytic processing of growth factors and turnover of extracellular matrix. These various biological activities are linked in part to the stoichiometry of tissue inhibitor of metalloprotein/matrix metalloprotein (TIMP/MMP)/surface protein interactions. TIMP-1, a secreted protein, can be detected on the cell surface only through its interaction with surface-bound proteins. Proteins anchored by glycosylphosphatidylinositol (GPI), when purified and added to cells or tissues, are efficiently incorporated into their surface membranes. A GPI anchor was fused to TIMP-1 to focus defined concentrations of the inhibitory protein independently on the surface of primary dermal fibroblast cells. Exogenously added recombinant TIMP-1-GPI effectively inserted into the cell membrane of fibroblasts blocked the secretion of MMPs and markedly altered the stoichiometry of MMP association with the cell surface. TIMP-1-GPI treatment resulted in inhibition of fibroblast-reduced proliferation, and transiently reduced expression of fibrosis-associated genes. These effects were dose dependent. Treated cells also showed a more proapoptotic phenotype based on apoptotic assays and western blot analysis for apoptosis-associated protein expression. GPI-anchored TIMP-1 may represent a more effective version of the protein for use in therapeutic approaches to help control fibrosis and scar formation.
authors.affiliation: Medizinische Klinik und Poliklinik IV, Campus Innenstadt, Klinikum der Ludwig-Maximilians-Universität München, Munich, Germany.
authors.forename: Roghieh
authors.initials: R
authors.lastname: Djafarzadeh
authors.affiliation: Medizinische Klinik und Poliklinik IV, Campus Innenstadt, Klinikum der Ludwig-Maximilians-Universität München, Munich, Germany.
authors.forename: Susan
authors.initials: S
authors.lastname: Notohamiprodjo
authors.affiliation: Medizinische Klinik und Poliklinik IV, Campus Innenstadt, Klinikum der Ludwig-Maximilians-Universität München, Munich, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Rieth
authors.affiliation: Medizinische Klinik und Poliklinik IV, Campus Innenstadt, Klinikum der Ludwig-Maximilians-Universität München, Munich, Germany.
authors.forename: Monika
authors.initials: M
authors.lastname: Hofstetter
authors.affiliation: Institute of Molecular Immunology, Helmholtz Zentrum München, Munich, Germany.
authors.forename: Elfriede
authors.initials: E
authors.lastname: Noessner
authors.affiliation: Medizinische Klinik und Poliklinik IV, Campus Innenstadt, Klinikum der Ludwig-Maximilians-Universität München, Munich, Germany. Electronic address: peter.nelson@med.uni-muenchen.de.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Nelson
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002921:Cicatrix / Q000503:physiopathology*; D020405:Dermis / Q000473:pathology*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005355:Fibrosis; D058851:GPI-Linked Proteins / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D020782:Matrix Metalloproteinases / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D058851:GPI-Linked Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D014409:Tumor Necrosis Factor-alpha; D020782:Matrix Metalloproteinases