HMGB1 : regulation of inflammatory functions and therapeutic blockade
Author: Lundbäck, Peter
Date: 2015-04-24
Location: CMM Lecture Hall, L8:00, Karolinska Sjukhuset, Solna
Time: 09.00
Department: Inst för medicin, Solna / Dept of Medicine, Solna
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Thesis (1.448Mb)
Abstract
The intracellular protein High Mobility Group Box Protein 1 (HMGB1) has been
identified as a pivotal mediator of inflammation. HMGB1 can be released by various
mechanisms and as an inflammatory mediator it induces both migration of
inflammatory cells and cytokine production. Consequently, HMGB1 has been
demonstrated to contribute to pathology in several inflammatory conditions.
Increasing evidence indicate that HMGB1 post-translational modifications (PTMs)
regulate both secretion and function of HMGB1.
The focus of this thesis work has been to investigate how selected PTMs regulate HMGB1 function and release and to define the presence of such modifications on HMGB1 in synovial fluid from patients with juvenile idiopathic arthritis (JIA). Furthermore, I have set the basis for HMGB1-blockade as a clinical treatment option by generating and characterizing the first known chimeric, humanized anti-HMGB1 antibody.
To examine the impact of redox-dependent PTMs on HMGB1 function, we first generated several cysteine redox isoforms of HMGB1. We found that all cysteines residues (C23, C45 and C106) required a defined redox state. A disulfide bridge between C23 and C45 with a concomitant C106 thiol was necessary for HMGB1 mediated cytokine-induction. In this disulfide redox isoform, HMGB1 activates TLR4. Furthermore, I have studied PTMs and their impact on HMGB1 secretion. We demonstrated that NLRC4 inflammasome activation induces hyperacetylation of key lysine stretches known to be associated with HMGB1 secretion, independently of priming signals. Addition of a priming signal induced reactive oxygen species (ROS) that stimulated a structural transition of HMGB1 to its cytokine-inducing, disulfide form. Hyperacetylated HMGB1 correlated significantly with inflammatory HMGB1 redox isoforms in joint fluid from JIA patients, indicating that HMGB1 is actively secreted during JIA and possesses inflammatory properties.
In addition, I recorded beneficial effects of mouse monoclonal anti-HMGB1 antibody (m2G7) treatment in experimental arthritis and in acetaminopheninduced liver injury. Importantly, I could demonstrate that a partly humanized version of the antibody (h2G7) retained its in vitro properties and in vivo therapeutic effects.
In conclusion, this thesis has significantly increased the understanding of the regulation of HMGB1 secretion and function during inflammation. The generation of an anti-HMGB1 chimeric antibody is an important step in development of a clinical anti-HMGB1 treatment.
The focus of this thesis work has been to investigate how selected PTMs regulate HMGB1 function and release and to define the presence of such modifications on HMGB1 in synovial fluid from patients with juvenile idiopathic arthritis (JIA). Furthermore, I have set the basis for HMGB1-blockade as a clinical treatment option by generating and characterizing the first known chimeric, humanized anti-HMGB1 antibody.
To examine the impact of redox-dependent PTMs on HMGB1 function, we first generated several cysteine redox isoforms of HMGB1. We found that all cysteines residues (C23, C45 and C106) required a defined redox state. A disulfide bridge between C23 and C45 with a concomitant C106 thiol was necessary for HMGB1 mediated cytokine-induction. In this disulfide redox isoform, HMGB1 activates TLR4. Furthermore, I have studied PTMs and their impact on HMGB1 secretion. We demonstrated that NLRC4 inflammasome activation induces hyperacetylation of key lysine stretches known to be associated with HMGB1 secretion, independently of priming signals. Addition of a priming signal induced reactive oxygen species (ROS) that stimulated a structural transition of HMGB1 to its cytokine-inducing, disulfide form. Hyperacetylated HMGB1 correlated significantly with inflammatory HMGB1 redox isoforms in joint fluid from JIA patients, indicating that HMGB1 is actively secreted during JIA and possesses inflammatory properties.
In addition, I recorded beneficial effects of mouse monoclonal anti-HMGB1 antibody (m2G7) treatment in experimental arthritis and in acetaminopheninduced liver injury. Importantly, I could demonstrate that a partly humanized version of the antibody (h2G7) retained its in vitro properties and in vivo therapeutic effects.
In conclusion, this thesis has significantly increased the understanding of the regulation of HMGB1 secretion and function during inflammation. The generation of an anti-HMGB1 chimeric antibody is an important step in development of a clinical anti-HMGB1 treatment.
List of papers:
I. Redox modification of cysteine residues regulates the cytokine activity of high mobility group box-1 (HMGB1). Huan Yang, Peter Lundbäck, Lars Ottosson, Helena Erlandsson-Harris, Emilie Venereau, Marco E. Bianchi, Yousef Al-Abed, Ulf Andersson, Kevin J. Tracey, Daniel J. Antoine Molecular Medicine. 2012 Mar 30;18:250-9
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. TLR activation regulates damage-associated molecular pattern isoforms released during pyroptosis. Sanna Nyström, Daniel Antoine, Peter Lundbäck, John G. Lock, Andreia F. Nita, Kari Högstrand, Alf Grandien, Helena Erlandsson-Harris, Ulf Andersson, Steven E. Applequist EMBO Journal. 2013 Jan 9;32(1):86-99
Fulltext (DOI)
Pubmed
View record in Web of Science®
III. Characterization of the inflammatory properties of actively released HMGB1 in juvenile idiopathic arthritis. Peter Lundbäck, Pernilla Stridh, Lena Klevenvall, Rosalind E. Jenkins, Marie Fischer, Erik Sundberg, Ulf Andersson, Daniel J. Antoine, Helena Erlandsson Harris Antioxidants&Redox Signaling. 2014 Dec 22.
Fulltext (DOI)
Pubmed
IV. Monoclonal anti-HMGB1 (high mobility group box chromosomal protein 1) antibody protection in two experimental arthritis models. Hanna Schierbeck, Peter Lundbäck, Karin Palmblad, Lena Klevenvall, Helena Erlandsson-Harris, Ulf Andersson, Lars Ottosson Molecular Medicine. 2011 Sep-Oct;17(9-10):1039-44
Fulltext (DOI)
Pubmed
View record in Web of Science®
V. Successful therapeutic HMGB1 neutralization using a humanized, chimeric monoclonal antibody. Peter Lundbäck, Jonathan D. Lea, Agnieszka Sowinska, Lars Ottosson, Camilla Melin-Fürst, Johanna Steen, Joanna I. Clarke, Anja Kipar, Lena Klevenvall, Huan Yang, Karin Palmblad, B. Kevin Park, Kevin J. Tracey, Anna Blom, Ulf Andersson, Daniel J. Antoine, and Helena Erlandsson Harris. [Manuscript]
I. Redox modification of cysteine residues regulates the cytokine activity of high mobility group box-1 (HMGB1). Huan Yang, Peter Lundbäck, Lars Ottosson, Helena Erlandsson-Harris, Emilie Venereau, Marco E. Bianchi, Yousef Al-Abed, Ulf Andersson, Kevin J. Tracey, Daniel J. Antoine Molecular Medicine. 2012 Mar 30;18:250-9
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. TLR activation regulates damage-associated molecular pattern isoforms released during pyroptosis. Sanna Nyström, Daniel Antoine, Peter Lundbäck, John G. Lock, Andreia F. Nita, Kari Högstrand, Alf Grandien, Helena Erlandsson-Harris, Ulf Andersson, Steven E. Applequist EMBO Journal. 2013 Jan 9;32(1):86-99
Fulltext (DOI)
Pubmed
View record in Web of Science®
III. Characterization of the inflammatory properties of actively released HMGB1 in juvenile idiopathic arthritis. Peter Lundbäck, Pernilla Stridh, Lena Klevenvall, Rosalind E. Jenkins, Marie Fischer, Erik Sundberg, Ulf Andersson, Daniel J. Antoine, Helena Erlandsson Harris Antioxidants&Redox Signaling. 2014 Dec 22.
Fulltext (DOI)
Pubmed
IV. Monoclonal anti-HMGB1 (high mobility group box chromosomal protein 1) antibody protection in two experimental arthritis models. Hanna Schierbeck, Peter Lundbäck, Karin Palmblad, Lena Klevenvall, Helena Erlandsson-Harris, Ulf Andersson, Lars Ottosson Molecular Medicine. 2011 Sep-Oct;17(9-10):1039-44
Fulltext (DOI)
Pubmed
View record in Web of Science®
V. Successful therapeutic HMGB1 neutralization using a humanized, chimeric monoclonal antibody. Peter Lundbäck, Jonathan D. Lea, Agnieszka Sowinska, Lars Ottosson, Camilla Melin-Fürst, Johanna Steen, Joanna I. Clarke, Anja Kipar, Lena Klevenvall, Huan Yang, Karin Palmblad, B. Kevin Park, Kevin J. Tracey, Anna Blom, Ulf Andersson, Daniel J. Antoine, and Helena Erlandsson Harris. [Manuscript]
Institution: Karolinska Institutet
Supervisor: Erlandsson Harris, Helena
Issue date: 2015-04-01
Rights:
Publication year: 2015
ISBN: 978-91-7549-853-9
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