DOCSLIB.ORG

Neutrophil Collagenase (Matrixmetalloproteinase-8): Regulatory Roles in Inflammationand Autoimmunity

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Neutrophil Collagenase (Matrixmetalloproteinase-8): Regulatory Roles in Inflammationand Autoimmunity NEUTROPHIL COLLAGENASE (MATRIX METALLOPROTEINASE-8): REGULATORY ROLES IN INFLAMMATION AND AUTOIMMUNITY by Jennifer H. Cox B.Sc., University of Victoria, 2001 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES (Biochemistry and Molecular Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) October 2008 © Jennifer H. Cox, 2008 ABSTRACT Inflammation is an essential process in wound healing and for the elimination of invading pathogens. However, unregulated inflammation can lead to numerous pathologies including autoimmu nities, tu morigenesis, and atherosclerosis. Matrix metalloproteinases (MMPs), once thought to be only extracellular matrix degrading enzymes, are now known to be key regulators of inflammatory and immune responses through proteolysis of bioactive molecules. MMP-8, a neutrophil-specific MMP, is protective in skin cancer models where MMP-8 knockout mice have an initial delay in neutrophil infiltration followed by a massive accumulation at the site of treatment. We investigated this delay in a murine air pouch model of acute inflammation, where MMP-8 deficiency caused decreased neutrophil migration in response to LPS. This was attributed to MMP-8 processing and activation of LPS-inducible CXC chemokine (LIX), a murine neutrophil chemoattractant. Indeed, MMP-8 knockout mice had normal neutrophil infiltration in response to synthetic analogs of cleaved LIX. Furthermore, homologous pathways with human chemokines CXCL5 and CXCL8 were described. In vivo, an indirect interaction between MMP-8 and LIX also occurs, whereby MMP-8 processes and inactivates cLl-proteinase inhibitor causing increased neutrophil elastase activity, which then efficiently cleaves and activates LIX. MMP-8 was protective in a model of rheumatoid arthritis where synovial tissues from MMP-8 deficient mice had an abundance of neutrophils. This prolonged neutrophil accumulation correlated with a loss of caspase-1 1 expression, consequent decreased caspase-3 activity and reduced apoptosis. MMP-8 shedding of TNF-a was also decreased in MMP-8 deficient leukocytes, potentially dampening a key apoptotic pathway in neutrophils. The role of MMPs in processing the Thi cell CXCR3-binding chemokines CXCL9, CXCL1O, and CXCL11 was investigated. The leukocytic MMPs -7, -8, -9, and -12 cleaved CXCL11 at both the amino and carboxy terminus. N-terminal cleavage resulted in the conversion of a receptor agonist to antagonist whereas C-terminal cleavage by MMP-8 caused a significant loss in glycosaminoglycan binding, demonstrating for the first time that direct chemokine proteolysis can regulate the formation of haptotactic gradients. Therefore, MMP-8 is a pivotal regulator in the onset and termination of inflammation, and has multifaceted roles in innate and acquired immunity as well as the autoimmune disorder rheumatoid arthritis. TABLE OF CONTENTS ABSTRACT ii TABLE OF CONTENTS iii LIST OF TABLES vi LIST OF FIGURES vii LIST OF ABBREVIATIONS ix ACKNOWLEDGEMENTS x CO-AUTHORSHIP STATEMENT xi CHAPTER 1. INTRODUCTION 1 The Basics of Matrix Metalloproteinases 1 The Neutrophil Collagenase: MMP-8 3 Cytokine Substrates: MMP Regulation of Inflammatory Signalling Molecules 5 Perspective 5 MMPs and Inflammation 6 Genetic Models of Inflammation 7 MMP Regulation of Cytokines 10 MMP Regulation of Chemokines 17 Future Perspectives 22 Themes and Hypotheses 23 References 25 CHAPTER 2. LPS RESPONSIVENESS AND NEUTROPHIL CHEMOTAXIS 3 IN VIVO REQUIRE PMN MMP-8 ACTIVITY Perspective 35 Introduction 36 Materials and Methods 39 Results 41 III Discussion 54 References 58 CHAPTER 3. MATRIX METALLOPROTEINASE PROCESSING OF CXCL1 1/I-TAG RESULTS IN LOSS OF CHEMOATTRACTANT ACTIVITY 62 AND ALTERED GLYCOSAMINOGLYCAN BINDING Perspective 62 Introduction 63 Materials and Methods 65 Results 70 Discussion 87 References 94 CHAPTER 4. MMP-8 DEFICIENCY DELAYS NEUTROPHIL APOPTOSIS AND EXACERBATES RHEUMATOID ARTHRITIS Perspective gg Introduction 100 Materials and Methods 102 Results 106 Discussion 126 References 131 CHAPTER 5. CONCLUSIONS AND PERSPECTIVES 136 References 143 APPENDIX A. PROCESSING OF ELR+ CHEMOKINES BY 144 NEUTROPHIL SERINE PROTEASES Perspective 144 Results 145 Summary and Future Experiments 154 References 156 iv APPENDIX B. PROTEOMIC ANALYSIS OF MMP-8 CLEAVAGE SPECIFICITY AND SUBSTRATE IDENTIFICATION 157 Perspective 157 Results 158 Summary and Future Experiments 162 References 164 APPENDIX C. UBC RESEARCH ETHICS BOARD CERTIFICATES 165 V LIST OF TABLES Table 1.1. Extracellular matrix and bioactive substrates of MMP-8 4 Table 1.2. Inflammatory and immune phenotypes of MMP-deficient mice 8 Bioactive MMP substrates: The pro-inflammatory and Table 1.3. 12 anti-inflammatory proteolysis events by MMPs vi LIST OF FIGURES Figure 1.1. Schematic structure of MMPs 2 MMP proteolysis of cytokines and chemokines to precisely control Fl g ure 1 2 15 the induction and termination of inflammation Figure 2.1. Impaired PMN responsiveness to LPS in MMP-8 deficient mice 42 Figure 2.2. MMP-8 cleavage of LIX 44 Figure 2.3. LIX is selectively cleaved by multiple MMPs 46 Figure 2.4. In vitro cellular responses to MMP-8 cleaved LIX 47 MMP-8 is required for PMN chemotaxis torard LIX in vivo, but is not Fi g ure 2 5 required for PMN cell migration Figure 2.6. MMP proccessing of CXCL8 and CXCL5 50 Figure 2.7. Enhanced bioactivity of MM P-truncated CXCLS in vitro and in vivo 53 Figure 3.1. MMP processing of CXCR3 ligands 71 Figure 3.2. MMP processing of murine CXCL1 I 74 MMP cleavage sites in CXCL9, CXCL1 0, CXCL1 1 and murine Fi g ure 3 3 CXCLI1 75 Figure 3.4. Titration of MMP processing of CXCLI I 77 Figure 3.5. Exosite interactions with CXCLI 1 79 MMP-mediated cleavages of CXCL1 1 result in loss of agonism and Fi gure 3 6 the generation of antagonism 80 Chemotactic migration is decreased and antagonized in response to Fl g ure 3 7 82 CXCL1 1 (5-73) and (5-58) Figure 3.8. Structural analysis of full length and truncated CXCL1 1 83 Figure 3.9. Heparin binding is altered upon CXCL1I cleavage 85 Model representing the potential role of leukocytic MMPs 7, 8, 9, and Fi g ure 3 10 88 12 in disrupting the haptotactic gradient of CXCL1 1 Figure 4.1. Generation and characterization of MRLJIprx Mmp8-I- mice 107 Figure 4.2. Increased joint swelling in male MMP-8 deficient MRL/lpr mice 109 vii Histological analysis of joints reveals increase neutrophil infiltration Fi g ure 4 3 112 in male MMP-8 deficient mice CLIP-CHIP analysis of bone marrow neutrophils from MMP-8 Fi g ure 44 115 . knockout and wild-type MRL/ipr mice Neutrophils and mononuclear cells from MMP-8 deficient mice have Fi g ure 4 5 117 abolished constitutive and inducible levels of caspase-1 1 MMP-8 deficient neutrophils have unaltered caspase-1 activity and Figure 4.6. 119 IL-I I secretion MMP-8 mediated cleavage of proTNF-a and shedding of TN F-cL from Figure 4.7. 122 neutroph ils Reduced caspase-3 activity and TUNEL staining Figure 4 in MMP-8 knockout 8 neutrophils 124 Proposed mechanisms for MMP-8 promotion of programmed cell Figure 4.9. death through induction of caspase-Il expression and shedding of 125 TNF-a Solving the MMP-8 puzzle: diverse functions of MMP-8 in Fi gure 5 . 1 140 . inflammation and acquired immunity Figure A.1. Neutrophil processing of LIX is independent of MMP-8 146 Figure A.2. Neutrophil-derived serine proteases cleave LIX 147 Figure A.3. Neutrophil elastase processing of murine ELR chemokines 149 Figure A.4. Neutrophil elastase processing of human ELR chemokines 151 Figure A.5. MMP-8 cleavage of cLl-proteinase inhibitor 152 Reduced LIX processing and increased full length al-PI in BALF Figure A.6. 153 from MMP-8 deficient mice Proposed mechanism for MMP-8 regulation of neutrophil elastase- Fi gure A 7 155 mediated LIX activation Figure B.1. Cleavage site specificities of murine and human MMP-8 159 CLIP-TAILS analysis of Mmp8-I- MEF secretome in the presence Fi g ure B 2 163 and absence of murine MMP-8 Viii LIST OF ABBREVIATIONS al-PI al-proteinase inhibitor AEBSF 2-aminoethyl benzenesulfonyl fluoride hydrochloride ADAM a disintigrin and metalloproteinase APMA 4-aminophenylmercuric acetate BALF bronchoalveolar lavage fluid BSA bovine serum albumin CATG cathepsin G CD circular dichroism DMEM dulbecco’s modified essential medium DMSO dimethyl sulfoxide DTT dithiothreitol GRO growth related oncogene HBSS Hank’s balanced salt solution ICE interleu kin-I converting enzyme IL interleukin IP-lO inducible protein 10 l-TAC interferon-gamma inducible T cell alpha chemoattractant LCD linker hemopexin C domain LIX LPS-inducible CXC chemokine LPS lipopolysaccharide MALDI-TOF matrix-assisted laser desorption ionization-time of flight MCP monocyte chemoattractant protein MEF murine embryonic fibroblast MIG monokine induced by gamma interferon MMP matrix metalloproteinase MS mass spectrometry MS/MS tandem mass spectrometry MT membrane type NE neutrophil elastase PAGE polyacrylamide gel electrophoresis PBS phosphate-buffered saline PMA phorbol 12-myristate 13-acetate PMN polymorphonuclear cell RPMI Roswell Park Memorial Institute medium SDF stromal derived factor SDS sodium dodecyl sulfate SLPI secreted leukocyte protease inhibitor TACE TNF alpha converting enzyme TGF transforming growth factor TIMP tissue inhibitor of metalloproteinases TNF tumor necrosis factor ix ACKNOWLEDGEMENTS First and foremost, I would like to thank Dr. Chris Overall for his optimism, encouragement, and genuine enthusiasm for science. I could not have imagined a more supportive supervisor, who always had confidence in me and truly gave me every opportunity to succeed. Also, I would like to acknowledge my supervisory committee for their involvement in this research: Dr. Francois Jean, Dr. Douglas Waterfield, and Dr. Leonard Foster. Furthermore, I would like to give a special thanks to collaborators Dr. Clive Roberts and Rendi Yan for their valuable contributions to this research.
Load more

Recommended publications
  • Structure-Function Relationship of Human Neutrophil Collagenase
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 2569-2573, April 1993 Biochemistry Structure-function relationship of human neutrophil collagenase: Identification of regions responsible for substrate specificity and general proteinase activity (matrix metalloproteinase/stromelysin/extracellular macromolecule) TOMOHIKO HIROSE*, CHRISTY PATTERSON*, TAYEBEH POURMOTABBEDt, CARLO L. MAINARDIt, AND KAREN A. HASTY*t Departments of *Anatomy and Neurobiology, and of tMedicine, University of Tennessee, Memphis, TN 38163 Communicated by Jerome Gross, December 16, 1992 (received for review September 10, 1992) ABSTRACT The family of matrix metalloproteinases is a identity and 70% chemical similarity with human neutrophil family ofclosely related enzymes that play an inportant role in collagenase (MMP-8; NC). Preliminary data from other lab- physiological and pathological processes of matrix degrada- oratories have pointed toward the COOH-terminal domain as tion. The most distinctive characteristic of interstitial collage- a major determinant in substrate specificity. Windsor et al. nases (fibroblast and neutrophil collagenases) is their ability to (16) have demonstrated that the cysteine residue immediately cleave interstitial coilagens at a single peptide bond; however, adjacent to the COOH terminus was essential for colla- the precise region of the enzyme responsible for this substrate genolytic activity of fibroblast collagenase (FC). Recently, specificity remains to be defined. To address this question, we Murphy et al. (17) have suggested that both COOH- and generated truncated mutants of neutrophil collagenase with NH2-terminal domains ofthe active enzyme contribute to the various deletions in the COOH-terminal domain and chimeric substrate specificity of collagenase and proposed a possible molecules between neutrophil collagenase and stromelysin and role of the COOH-terminal domain in substrate binding.
    [Show full text]
  • Inflammation and Extracellular Matrix Degradation Mediated by Activated Transcription Factors Nuclear Factor-Κb and Activator P
    American Journal of Pathology, Vol. 166, No. 6, June 2005 Copyright © American Society for Investigative Pathology Matrix Pathobiology Inflammation and Extracellular Matrix Degradation Mediated by Activated Transcription Factors Nuclear Factor-␬B and Activator Protein-1 in Inflammatory Acne Lesions in Vivo Sewon Kang, Soyun Cho, Jin Ho Chung, Despite its common occurrence, the pathogenesis of Craig Hammerberg, Gary J. Fisher, and acne is not fully understood. Excessive shedding of ep- John J. Voorhees ithelial cells from the walls of follicles combined with increased amounts of sebum produced by associated From the Department of Dermatology, University of Michigan Medical School, Ann Arbor, Michigan sebaceous glands are two important factors that contrib- ute to follicular obstruction.4,5 This obstruction leads to the formation of microcomedos, which are believed to be the precursor lesions of acne.5 These microcomedos may evolve into clinically visible comedos (blackheads Acne is the most common skin disease, causing sig- and whiteheads) or inflammatory lesions (papules, pus- nificant psychosocial problems for those afflicted. tules, or cysts). Propionibacterium acnes (P. acnes) are Currently available agents for acne treatment, such as gram-positive, anaerobic diphtheroids that are part of the oral antibiotics and isotretinoin (Accutane), have lim- 6 normal skin flora. In the generation of inflammatory le- ited use. Thus, development of novel agents to treat sions, proliferation of P. acnes in the obstructed follicles is this disease is needed. However, the pathophysiology believed to be critical.6,7 This is based on in vitro data that of acne inflammation is poorly understood. Before P. acnes stimulate monocytes to produce proinflamma- new therapeutic strategies can be devised, knowledge tory mediators and chemotactic factors,7,8 and time- regarding molecular mechanisms of acne inflamma- tested clinical observations that antibiotics that inhibit P.
    [Show full text]
  • Physical Journal Volume 79 October 2000 2138–2149
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector 2138 Biophysical Journal Volume 79 October 2000 2138–2149 pH- and Temperature-Dependence of Functional Modulation in Metalloproteinases. A Comparison between Neutrophil Collagenase and Gelatinases A and B Giovanni Francesco Fasciglione,* Stefano Marini,* Silvana D’Alessio,† Vincenzo Politi,† and Massimo Coletta* *Department of Experimental Medicine and Biochemical Sciences, University of Roma Tor Vergata, I-00133 Roma, and †PoliFarma, I-00155 Roma, Italy ABSTRACT Metalloproteases are metalloenzymes secreted in the extracellular fluid and involved in inflammatory patholo- gies or events, such as extracellular degradation. A Zn2ϩ metal, present in the active site, is involved in the catalytic mechanism, and it is generally coordinated with histidyl and/or cysteinyl residues of the protein moiety. In this study we have investigated the effect of both pH (between pH 4.8 and 9.0) and temperature (between 15°C and 37°C) on the enzymatic functional properties of the neutrophil interstitial collagenase (MMP-8), gelatinases A (MMP-2) and B (MMP-9), using the same Ϸ synthetic substrate, namely MCA-Pro-Leu-Gly Leu-DPA-Ala-Arg-NH2. A global analysis of the observed proton-linked behavior for kcat/Km, kcat, and Km indicates that in order to have a fully consistent description of the enzymatic action of these metalloproteases we have to imply at least three protonating groups, with differing features for the three enzymes investi- gated, which are involved in the modulation of substrate interaction and catalysis by the enzyme. This is the first investigation of this type on recombinant collagenases and gelatinases of human origin.
    [Show full text]
  • Peptides and Peptidomimetics As Inhibitors of Enzymes Involved in Fibrillar Collagen Degradation
    materials Review Peptides and Peptidomimetics as Inhibitors of Enzymes Involved in Fibrillar Collagen Degradation Patrycja Ledwo ´n 1,2 , Anna Maria Papini 3 , Paolo Rovero 2,* and Rafal Latajka 1,* 1 Department of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Science and Technology, 50-370 Wroclaw, Poland; [email protected] 2 Interdepartmental Research Unit of Peptide and Protein Chemistry and Biology, Department of Neurosciences, Psychology, Drug Research and Child Health-Section of Pharmaceutical Sciences and Nutraceutics, University of Florence, 50019 Sesto Fiorentino, Firenze, Italy 3 Interdepartmental Research Unit of Peptide and Protein Chemistry and Biology, Department of Chemistry “Ugo Schiff”, University of Florence, 50019 Sesto Fiorentino, Firenze, Italy; annamaria.papini@unifi.it * Correspondence: paolo.rovero@unifi.it (P.R.); [email protected] (R.L.) Abstract: Collagen fibres degradation is a complex process involving a variety of enzymes. Fibrillar collagens, namely type I, II, and III, are the most widely spread collagens in human body, e.g., they are responsible for tissue fibrillar structure and skin elasticity. Nevertheless, the hyperactivity of fibrotic process and collagen accumulation results with joints, bone, heart, lungs, kidneys or liver fibroses. Per contra, dysfunctional collagen turnover and its increased degradation leads to wound healing disruption, skin photoaging, and loss of firmness and elasticity. In this review we described the main enzymes participating in collagen degradation pathway, paying particular attention to enzymes degrading fibrillar collagen. Therefore, collagenases (MMP-1, -8, and -13), elastases, and cathepsins, together with their peptide and peptidomimetic inhibitors, are reviewed. This information, related Citation: Ledwo´n,P.; Papini, A.M.; to the design and synthesis of new inhibitors based on peptide structure, can be relevant for future Rovero, P.; Latajka, R.
    [Show full text]
  • Handbook of Proteolytic Enzymes Second Edition Volume 1 Aspartic and Metallo Peptidases
    Handbook of Proteolytic Enzymes Second Edition Volume 1 Aspartic and Metallo Peptidases Alan J. Barrett Neil D. Rawlings J. Fred Woessner Editor biographies xxi Contributors xxiii Preface xxxi Introduction ' Abbreviations xxxvii ASPARTIC PEPTIDASES Introduction 1 Aspartic peptidases and their clans 3 2 Catalytic pathway of aspartic peptidases 12 Clan AA Family Al 3 Pepsin A 19 4 Pepsin B 28 5 Chymosin 29 6 Cathepsin E 33 7 Gastricsin 38 8 Cathepsin D 43 9 Napsin A 52 10 Renin 54 11 Mouse submandibular renin 62 12 Memapsin 1 64 13 Memapsin 2 66 14 Plasmepsins 70 15 Plasmepsin II 73 16 Tick heme-binding aspartic proteinase 76 17 Phytepsin 77 18 Nepenthesin 85 19 Saccharopepsin 87 20 Neurosporapepsin 90 21 Acrocylindropepsin 9 1 22 Aspergillopepsin I 92 23 Penicillopepsin 99 24 Endothiapepsin 104 25 Rhizopuspepsin 108 26 Mucorpepsin 11 1 27 Polyporopepsin 113 28 Candidapepsin 115 29 Candiparapsin 120 30 Canditropsin 123 31 Syncephapepsin 125 32 Barrierpepsin 126 33 Yapsin 1 128 34 Yapsin 2 132 35 Yapsin A 133 36 Pregnancy-associated glycoproteins 135 37 Pepsin F 137 38 Rhodotorulapepsin 139 39 Cladosporopepsin 140 40 Pycnoporopepsin 141 Family A2 and others 41 Human immunodeficiency virus 1 retropepsin 144 42 Human immunodeficiency virus 2 retropepsin 154 43 Simian immunodeficiency virus retropepsin 158 44 Equine infectious anemia virus retropepsin 160 45 Rous sarcoma virus retropepsin and avian myeloblastosis virus retropepsin 163 46 Human T-cell leukemia virus type I (HTLV-I) retropepsin 166 47 Bovine leukemia virus retropepsin 169 48
    [Show full text]
  • Characterization of Staphylococcus Aureus Skin Infection Using a New in Vivo Proliferation Biosensor”
    Characterization of Staphylococcus aureus skin infection using a new in vivo proliferation biosensor Dissertation zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) genehmigt durch die Fakultät für Naturwissenschaften der Otto-von-Guericke-Universität Magdeburg von M. Sc. Elena Anne Seiß (geb. Haas) geb. am 14.03.1988 in Erkelenz Gutachter: Prof. Dr. Andreas J. Müller Prof. Dr. rer. nat. Christiane Wolz eingereicht am: 08.01.2019 verteidigt am: 26.06.2019 ABSTRACT ABSTRACT M. Sc. Elena A. Seiß (née Haas): “Characterization of Staphylococcus aureus skin infection using a new in vivo proliferation biosensor” Distinct bacterial growth rates are decisive for the outcome of infections: First, they impact on the pathogen's susceptibility to both immune effector mechanisms and antibiotic treatment, and can therefore contribute to persistence during chronic or relapsing infections. Second, fast-growing bacteria are a source of more, and different, pathogen-associated molecules than low-proliferating or inactive microbes, thus differentially shaping the immune response. It has however remained difficult to measure bacterial growth rates in the ongoing infection. Here, I established an in vivo biosensor for measuring proliferation of the Gram-positive bacterium Staphylococcus aureus (S. aureus) on a single cell-level in vivo. The method is based on the photoconvertible fluorescence protein mKikume, which can be photoconverted by violet light at any given time point, even during in vivo infection in a non- invasive way. This allows determining bacterial proliferation rates as a function of the recovery from photoconversion. Importantly, I could show that indeed bacterial proliferation, and not protein turnover in nonproliferating bacteria, is mainly responsible for the fluorescence recovery after photoconversion readout of the biosensor.
    [Show full text]
  • Collagenolytic and Gelatinolytic Matrix Metalloproteinases And
    British Journal of Cancer (2000) 82(3), 657–665 © 2000 Cancer Research Campaign DOI: 10.1054/ bjoc.1999.0978, available online at http://www.idealibrary.com on Collagenolytic and gelatinolytic matrix metalloproteinases and their inhibitors in basal cell carcinoma of skin: comparison with normal skin J Varani1, Y Hattori1, Y Chi1, T Schmidt1, P Perone1, ME Zeigler1, DJ Fader2 and TM Johnson2 Departments of 1Pathology and 2Dermatology, The University of Michigan Medical School, 1301 Catherine Road, PO Box 0602, Ann Arbor, MI 48109, USA Summary Tissue from 54 histologically-identified basal cell carcinomas of the skin was obtained at surgery and assayed using a combination of functional and immunochemical procedures for matrix metalloproteinases (MMPs) with collagenolytic activity and for MMPs with gelatinolytic activity. Collagenolytic enzymes included MMP-1 (interstitial collagenase), MMP-8 (neutrophil collagenase) and MMP-13 (collagenase-3). Gelatinolytic enzymes included MMP-2 (72-kDa gelatinase A/type IV collagenase) and MMP-9 (92-kDa gelatinase B/type IV collagenase). Inhibitors of MMP activity including tissue inhibitor of metalloproteinases-1 and -2 (TIMP-1 and TIMP-2) were also assessed. All three collagenases and both gelatinases were detected immunochemically. MMP-1 appeared to be responsible for most of the functional collagenolytic activity while gelatinolytic activity reflected both MMP-2 and MMP-9. MMP inhibitor activity was also present, and appeared, based on immunochemical procedures, to reflect the presence of TIMP-1 but not TIMP-2. As a group, tumours identified as having aggressive-growth histologic patterns were not distinguishable from basal cell carcinomas with less aggressive-growth histologic patterns.
    [Show full text]
  • The Rebirth of Matrix Metalloproteinase Inhibitors: Moving Beyond the Dogma
    cells Review The Rebirth of Matrix Metalloproteinase Inhibitors: Moving Beyond the Dogma Gregg B. Fields 1,2 1 Institute for Human Health & Disease Intervention, Department of Chemistry & Biochemistry, and the Center for Molecular Biology & Biotechnology, Florida Atlantic University, Jupiter, FL 33458, USA; fi[email protected]; Tel.: +1-561-799-8577 2 Department of Chemistry, The Scripps Research Institute/Scripps Florida, Jupiter, FL 33458, USA Received: 2 August 2019; Accepted: 26 August 2019; Published: 27 August 2019 Abstract: The pursuit of matrix metalloproteinase (MMP) inhibitors began in earnest over three decades ago. Initial clinical trials were disappointing, resulting in a negative view of MMPs as therapeutic targets. As a better understanding of MMP biology and inhibitor pharmacokinetic properties emerged, it became clear that initial MMP inhibitor clinical trials were held prematurely. Further complicating matters were problematic conclusions drawn from animal model studies. The most recent generation of MMP inhibitors have desirable selectivities and improved pharmacokinetics, resulting in improved toxicity profiles. Application of selective MMP inhibitors led to the conclusion that MMP-2, MMP-9, MMP-13, and MT1-MMP are not involved in musculoskeletal syndrome, a common side effect observed with broad spectrum MMP inhibitors. Specific activities within a single MMP can now be inhibited. Better definition of the roles of MMPs in immunological responses and inflammation will help inform clinic trials, and multiple studies indicate that modulating MMP activity can improve immunotherapy. There is a U.S. Food and Drug Administration (FDA)-approved MMP inhibitor for periodontal disease, and several MMP inhibitors are in clinic trials, targeting a variety of maladies including gastric cancer, diabetic foot ulcers, and multiple sclerosis.
    [Show full text]
  • A Novel Urinary Proteomics Classifier for Non-Invasive Evaluation
    proteomes Article A Novel Urinary Proteomics Classifier for Non-Invasive Evaluation of Interstitial Fibrosis and Tubular Atrophy in Chronic Kidney Disease Lorenzo Catanese 1,2,3 , Justyna Siwy 4,* , Emmanouil Mavrogeorgis 4,5, Kerstin Amann 6, Harald Mischak 4 , Joachim Beige 7,8,9 and Harald Rupprecht 1,2,3 1 Department of Nephrology, Angiology and Rheumatology, Klinikum Bayreuth GmbH, 95447 Bayreuth, Germany; [email protected] (L.C.); [email protected] (H.R.) 2 Kuratorium for Dialysis and Transplantation (KfH) Bayreuth, 95445 Bayreuth, Germany 3 Friedrich-Alexander-University Erlangen-Nürnberg, 91054 Erlangen, Germany 4 Mosaiques Diagnostics GmbH, 30659 Hannover, Germany; [email protected] (E.M.); [email protected] (H.M.) 5 Institute for Molecular Cardiovascular Research (IMCAR), RWTH Aachen University Hospital, 52074 Aachen, Germany 6 Department of Nephropathology, Institute of Pathology, University of Erlangen-Nürnberg, 91054 Erlangen, Germany; [email protected] 7 Department of Infectious Diseases/Tropical Medicine, Nephrology/KfH Renal Unit and Rheumatology, St. Georg Hospital Leipzig, 04129 Leipzig, Germany; [email protected] 8 Kuratorium for Dialysis and Transplantation (KfH) Renal Unit, Hospital St. Georg, 04129 Leipzig, Germany 9 Department of Internal Medicine II, Martin-Luther-University Halle/Wittenberg, Citation: Catanese, L.; Siwy, J.; 06108 Halle (Saale), Germany Mavrogeorgis, E.; Amann, K.; * Correspondence: [email protected] Mischak, H.; Beige, J.; Rupprecht, H. A Novel Urinary Proteomics Abstract: Non-invasive urinary peptide biomarkers are able to detect and predict chronic kidney Classifier for Non-Invasive disease (CKD). Moreover, specific urinary peptides enable discrimination of different CKD etiologies Evaluation of Interstitial Fibrosis and and offer an interesting alternative to invasive kidney biopsy, which cannot always be performed.
    [Show full text]
  • Chapter 3: Endogenous Collagenase
    Open Access A Historical Review of Enzymatic Debridement: Revisited Chapter 3: Endogenous Collagenase The role of matrix metalloproteinases in wound repair The serine proteinases comprise the largest family of extracellular enzymes and include plasmin, plasminogen activators, and leukocyte elastase, as well as the coagulation and digestive proteinases. Generally, these are potent enzymes with broad catalytic speciicity and are readily available when needed. Plasminogen, the inactive form of plasmin, is present in high concentrations in blood and tissue. Neutrophils store an abundance of leukocyte elastase. In contrast, the metalloproteinases in wounded tissues have more deined substrate speciicity and are generally produced on demand. The structural and functional diversity of matrix metalloproteinases (MMPs) rivals that of the superfamily of collagens (reviewed in the previous chapter). The MMPs belong to a large family of zinc-dependent endopeptidases, the irst of which was described over a half century ago. MMPs were discovered initially as the agents responsible for tail resorption during frog metamorphosis [1-3] and have since been identiied as the main processors of extracellular matrix (ECM) components [4]. MMPs have also been implicated in more sophisticated processes than mere ECM turnover [5,6]. These include the activation or inactivation of other proteins through limited proteolysis of selected bonds, as well as the shedding of membrane- anchored forms into circulation. Substrates include other (pro-)proteases, protease inhibitors, clotting factors, antimicrobial peptides, chemotactic and adhesion molecules, and growth factors, hormones, cytokines, as well as, their receptors and binding proteins. In such shedding functions, MMPs overlap in substrate speciicity, and in spatial and temporal location [4,7,8].
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2011/0044968 A1 Bolotin Et Al
    US 20110044968A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0044968 A1 Bolotin et al. (43) Pub. Date: Feb. 24, 2011 (54) COMPOSITIONS FOR TREATMENT WITH (86). PCT No.: PCT/USO9A36648 METALLOPEPTIDASES, METHODS OF MAKING AND USING THE SAME S371 (c)(1), (2), (4) Date: Nov. 5, 2010 (75) Inventors: Elijah M. Bolotin, Bothell, WA Related U.S. Application Data WA(US); (US); Gerardo Penelope Castillo, Markham, Bothell, (60) Fyal application No. 61/068.896, filed on Mar. Clifton, VA (US); Manshun Lai, s Bothell, WA (US) Publication Classification (51) Int. Cl. Correspondence Address: A638/54 (2006.01) WILSON SONSIN GOODRCH AND ROSAT f CI2N 9/96 (2006.01) PHARMAN LTD A6IP3L/00 (2006.01) 650 PAGE MILL ROAD A6IP 25/28 (2006.01) PALO ALTO, CA 94.304 (US) (52) U.S. Cl. ........................................ 424/94.3; 435/188 (57) ABSTRACT (73)73) AssigneeAssi : WPh re NC orporation,tion. SeattlSealue, The present invention is directed to biocompatible composi tions and the use of metal bridges to connect a back-bone and a metallopeptidase active agent. In certain instances, the Sub (21) Appl. No.: 12/921,670 ject compositions provide a means of achieving Sustained release of the metallopeptidase active agent after administra (22) PCT Filed: Mar. 10, 2009 tion to a subject. Patent Application Publication Feb. 24, 2011 Sheet 1 of 13 US 2011/0044968 A1 Figure 1 (1) Polymeric Backbone (branched or unbranched) 2) Chelating moieties (3) Metalions ( 4) Protective chains (5) Metallopeptidase active agent Patent Application Publication Feb. 24, 2011 Sheet 2 of 13 US 2011/0044968 A1 Figure 2 O 120% poss C D 100% O 80% w 60% s CD 20% n S PGC-NTA-Zn PGC-NTA Lysostaphin Lysostaphin Patent Application Publication Feb.
    [Show full text]
  • Biochem Cv 1/1/96
    THE UNIVERSITY OF BRITISH COLUMBIA Curriculum Vitae for Faculty Members Date: May 14, 2014 1. SURNAME: Overall FIRST NAME: Christopher MIDDLE NAME: Mark 2. DEPARTMENTS: Oral Biological and Medical Sciences Associate Member Biochemistry & Molecular Biology AFFILIATIONS: U.B.C. Life Sciences Institute U.B.C. Centre for Blood Research The Vancouver Prostate Centre U.B.C. Centre for Drug Research and Development U.B.C. Genome Science and Technology Graduate Program Fraser Health Authority affiliated researcher Canadian National Proteomics Network British Columbian Proteomics Network Australian Protease Network CONTACT: 4.401 Life Sciences Institute, 2350 Health Sciences Mall Vancouver, BC V6T 1Z3, Canada +1-604-822-2958, [email protected], www.clip.ubc.ca 3. FACULTY: Dentistry 4. PRESENT RANK: Full Professor SINCE: 01/07/00 Tier I Canada Research Chair in Proteinase Proteomics and Systems Biology SINCE: 01/01/01 Honorary Professor, Institute of Molecular Medicine and Cell Research, Albert-Ludwigs Universität Freiburg SINCE: 1/05/14 Short Biographical Sketch Dr. Overall is a Professor and Tier 1 Canada Research Chair in Proteinase Proteomics and Systems Biology at the University of British Columbia, Centre for Blood Research. He completed his undergraduate, Honors Science and Masters degrees at the University of Adelaide, South Australia; his Ph.D. in Biochemistry at the University of Toronto; and was a MRC Centennial Fellow in his post-doctoral work with Dr. Michael Smith, Nobel Laureate, Biotechnology Laboratory, University of British Columbia. On Sabbatical in 1997-1998 he was a Visiting Senior Scientist at British Biotech Pharmaceuticals, Oxford, UK and in 2004/2008 he was a Visiting Senior Scientist at the Expert Protease Platform, Novartis Pharmaceuticals, Basel, Switzerland where he learned industrial scale concepts and the science of drug development.
    [Show full text]