title: Early Secretory Antigenic Target-6 Drives Matrix Metalloproteinase-10 Gene Expression and Secretion in Tuberculosis.
abstract: Tuberculosis (TB) causes disease worldwide, and multidrug resistance is an increasing problem. Matrix metalloproteinases (MMPs), particularly the collagenase MMP-1, cause lung extracellular matrix destruction, which drives disease transmission and morbidity. The role in such tissue damage of the stromelysin MMP-10, a key activator of the collagenase MMP-1, was investigated in direct Mycobacterium tuberculosis (Mtb)-infected macrophages and in conditioned medium from Mtb-infected monocyte-stimulated cells. Mtb infection increased MMP-10 secretion from primary human macrophages 29-fold, whereas Mtb-infected monocytes increased secretion by 4.5-fold from pulmonary epithelial cells and 10.5-fold from fibroblasts. Inhibition of MMP-10 activity decreased collagen breakdown. In two independent cohorts of patients with TB from different continents, MMP-10 was increased in both induced sputum and bronchoalveolar lavage fluid compared with control subjects and patients with other respiratory diseases (both P < 0.05). Mtb drove 3.5-fold greater MMP-10 secretion from human macrophages than the vaccine strain bacillus Calmette-Guerin (P < 0.001), whereas both mycobacteria up-regulated TNF-α secretion equally. Using overlapping, short, linear peptides covering the sequence of early secretory antigenic target-6, a virulence factor secreted by Mtb, but not bacillus Calmette-Guerin, we found that stimulation of human macrophages with a single specific 15-amino acid peptide sequence drove threefold greater MMP-10 secretion than any other peptide (P < 0.001). Mtb-driven MMP-10 secretion was inhibited in a dose-dependent manner by p38 and extracellular signal-related kinase mitogen-activated protein kinase blockade (P < 0.001 and P < 0.01 respectively), but it was not affected by inhibition of NF-κB. In summary, Mtb activates inflammatory and stromal cells to secrete MMP-10, and this is partly driven by the virulence factor early secretory antigenic target-6, implicating it in TB-associated tissue destruction.
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.|2 Centre for Inflammation and Tissue Repair, Respiratory Medicine, University College London, London, United Kingdom.
authors.forename: Sara
authors.identifier: http://orcid.org/0000-0003-2608-4444
authors.initials: S
authors.lastname: Brilha
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.
authors.forename: Tarangini
authors.initials: T
authors.lastname: Sathyamoorthy
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.
authors.forename: Laura H
authors.initials: LH
authors.lastname: Stuttaford
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.|3 Clinical Infectious Diseases Research Initiative, Institute of Infectious Diseases and Molecular Medicine, University of Cape Town, Cape Town, South Africa.|4 Department of Clinical Research, London School of Hygiene and Tropical Medicine, London, United Kingdom.
authors.forename: Naomi F
authors.initials: NF
authors.lastname: Walker
authors.affiliation: 3 Clinical Infectious Diseases Research Initiative, Institute of Infectious Diseases and Molecular Medicine, University of Cape Town, Cape Town, South Africa.|5 Department of Medicine, Imperial College London, London, United Kingdom.|6 The Francis Crick Institute, London, United Kingdom.|7 Wellcome Trust Imperial College Centre for Global Health, Imperial College London, London, United Kingdom; and.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Wilkinson
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.
authors.forename: Shivani
authors.initials: S
authors.lastname: Singh
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.
authors.forename: Rachel C
authors.initials: RC
authors.lastname: Moores
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.|8 National Institute of Health Research Respiratory Biomedical Research Unit, Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
authors.forename: Paul T
authors.initials: PT
authors.lastname: Elkington
authors.affiliation: 1 Infectious Diseases and Immunity, Imperial College London, London, United Kingdom.|7 Wellcome Trust Imperial College Centre for Global Health, Imperial College London, London, United Kingdom; and.
authors.forename: Jon S
authors.initials: JS
authors.lastname: Friedland
mesh_terms: D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008168:Lung / Q000473:pathology; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D053506:Matrix Metalloproteinase 10 / Q000235:genetics* / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000472:pathogenicity; D016328:NF-kappa B / Q000378:metabolism; D014376:Tuberculosis / Q000235:genetics* / Q000382:microbiology*; D032581:Tuberculosis Vaccines / Q000276:immunology; D014774:Virulence
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C093276:ESAT-6 protein, Mycobacterium tuberculosis; D016328:NF-kappa B; D032581:Tuberculosis Vaccines; D020928:Mitogen-Activated Protein Kinases; D053506:Matrix Metalloproteinase 10; D020781:Matrix Metalloproteinase 1
keywords: early secretory antigenic target-6; matrix metalloproteinases; tuberculosis