title: Combinatorial Screening Identifies Novel Promiscuous Matrix Metalloproteinase Activities that Lead to Inhibition of the Therapeutic Target IL-13.
abstract: The practical realization of disease modulation by catalytic degradation of a therapeutic target protein suffers from the difficulty to identify candidate proteases, or to engineer their specificity. We identified 23 measurable, specific, and new protease activities using combinatorial screening of 27 human proteases against 24 therapeutic protein targets. We investigate the cleavage of monocyte chemoattractant protein 1, interleukin-6 (IL-6), and IL-13 by matrix metalloproteinases (MMPs) and serine proteases, and demonstrate that cleavage of IL-13 leads to potent inhibition of its biological activity in vitro. MMP-8 degraded human IL-13 most efficiently in vitro and ex vivo in human IL-13 transgenic mouse bronchoalveolar lavage. Hence, MMP-8 is a therapeutic protease lead against IL-13 for inflammatory conditions whereby reported genetic and genomics data suggest an involvement of MMP-8. This work describes the first exploitation of human enzyme promiscuity for therapeutic applications, and reveals both starting points for protease-based therapies and potential new regulatory networks in inflammatory disease.
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune, Granta Park, Cambridge CB21 6GH, UK. Electronic address: urbachc@medimmune.com.
authors.forename: Carole
authors.initials: C
authors.lastname: Urbach
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune, Granta Park, Cambridge CB21 6GH, UK; Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK.
authors.forename: Nathaniel C
authors.initials: NC
authors.lastname: Gordon
authors.affiliation: Department of Respiratory, Inflammation and Autoimmunity, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Strickland
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: David
authors.initials: D
authors.lastname: Lowne
authors.affiliation: Department of Oncology, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Cathy
authors.initials: C
authors.lastname: Joberty-Candotti
authors.affiliation: Department of Respiratory, Inflammation and Autoimmunity, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: May
authors.affiliation: Non Clinical Biostatistics, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Athula
authors.initials: A
authors.lastname: Herath
authors.affiliation: Department of Dermatology, University Medical Center, 3508 GA Utrecht, the Netherlands.
authors.forename: DirkJan
authors.initials: D
authors.lastname: Hijnen
authors.affiliation: Department of Dermatology, University Medical Center, 3508 GA Utrecht, the Netherlands.
authors.forename: Judith L
authors.initials: JL
authors.lastname: Thijs
authors.affiliation: Department of Dermatology, University Medical Center, 3508 GA Utrecht, the Netherlands.
authors.forename: Carla A
authors.initials: CA
authors.lastname: Bruijnzeel-Koomen
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Ralph R
authors.initials: RR
authors.lastname: Minter
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK.
authors.forename: Florian
authors.initials: F
authors.lastname: Hollfelder
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Lutz
authors.initials: L
authors.lastname: Jermutus
mesh_terms: D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism; D003876:Dermatitis, Atopic / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D018793:Interleukin-13 / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D007700:Kinetics; D020784:Matrix Metalloproteinase 8 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D015202:Protein Engineering; D059748:Proteolysis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D018932:Chemokine CCL2; D018793:Interleukin-13; D015850:Interleukin-6; D011994:Recombinant Proteins; D020782:Matrix Metalloproteinases; D020784:Matrix Metalloproteinase 8