IDENTIFICATION OF MEMBRANE-TYPE 1 MATRIX METALLOPROTEINASE AS A NOVEL THERAPEUTIC TARGET OF RHEUMATOID ARTHRITIS

Lead Research Organisation: University of Oxford
Department Name: Kennedy Institute

Abstract

Rheumatoid arthritis (RA) is a systemic autoimmune disease whose key pathological features is destruction of joint tissues including cartilage. Our recent data indicated that a cell membrane-anchored protein degrading enzyme termed, MT1-MMP, plays an essential role in cartilage erosion in RA. We hypothesize that specific inhibition of MT1-MMP can be a potential RA treatment. We propose to address this by assessing the effect of MT1-MMP inhibition in RA progression in a collagen-induced arthritis (CIA) in transgenic mice that express a dominant negative inhibitor of MT1-MMP. We will also analyze the effect of a MT1-MMP-specific inhibitory antibody in mice with CIA. Completing the proposed work would establish MT1-MMP as a novel therapeutic target of RA treatment, which may lead into development of novel RA therapy in the future.

Technical Summary

Rheumatoid arthritis (RA) is a systemic autoimmune disease whose key pathological features is destruction of joint tissues by inflamed synovial pannus tissue leading to a loss of joint function. We have recently discovered that one of the membrane-anchored matrix metalloproteinases (MT-MMPs), MT1-MMP, plays an essential role in promoting pannus invasion into cartilage in RA. MT1-MMP is highly expressed in the synovial cells forming a cartilage invasion front and inhibition of MT1-MMP effectively abrogates synovial cell invasion into cartilage matrix. MT1-MMP has also been shown to be an important molecule in the process of angiogenesis, as well as monocyte endothelial transmigration, both of which are essential steps in RA progression. Therefore, we hypothesize that specific inhibition of MT1-MMP can be a potential therapeutic intervention for RA treatment. To test the effect of MT1-MMP inhibition in RA progression, we have developed useful tools: transgenic mice that express a dominant negative form of MT1-MMP in fibroblasts and an MT1-MMP-specific inhibitory antibody. We will challenge the transgenic mice with collagen-induced arthritis (CIA) to examine the role of MT1-MMP in synovial fibroblasts during development of arthritis in vivo. We will also administer the MT1-MMP inhibitory antibody to mice with CIA in order to examine the role of MT1-MMP in broader cell populations during RA development, including endothelial cells and macrophages. Completing the proposed work would establish MT1-MMP as a novel therapeutic target of RA treatment, which may lead into development of novel RA therapy in the future.

Publications

10 25 50
 
Description Autofluorescence lifetime metrology for label-free readouts of heart disease and arthritis
Amount £223,407 (GBP)
Funding ID EP/I02770X/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 04/2011 
End 12/2014
 
Description LAB282
Amount £516,431 (GBP)
Funding ID OUI16295 
Organisation University of Oxford 
Department Oxford University Innovation
Sector Private
Country United Kingdom
Start 02/2019 
End 02/2020
 
Description The role of cell surface metalloproteinases in the development of rheumatoid arthritis
Amount £300,000 (GBP)
Funding ID 19797 
Organisation Versus Arthritis 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2012 
End 09/2015
 
Title MT1-MMP highly selective inhibitor, E2_C6 
Description This inhibitor was developed by collaboration with Prof Gillian Murphy at University of Cambridge. Our involvement was to screen the candidate inhibitors. This is an biologic inhibitor screened by phage-display library. It inhibits MT1-MMP-depedent cellular invasion and ECM degradation. 
Type Of Material Biological samples 
Year Produced 2013 
Provided To Others? Yes  
Impact This is highly selective biological inhibitor. 
 
Title Method to activate DDRs with cartilage matrix 
Description Assay method to examine cartilage can stimulate DDR2 by fibroblasts 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Bovine nasal cartilage is sliced into 2 mm thickness and punched with biopsy punch with 4 mm diameter. The cartilage was placed in the 48 well plate and culture the cell atop of the tissue for 6h. Cartilage with cells are then transferred to new 48 well to remove cells that did not attached. Culture another 40h to see the down stream event of DDR2 activation. 
 
Title Soluble DDR2-Fc fusion protein 
Description This is a recombinant DDR2-Fc fusion protein. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact This is very useful to study interaction of DDR2 with collagen. 
 
Description DDR-selective RTK inhibitors 
Organisation Harvard University
Country United States 
Sector Academic/University 
PI Contribution We examine the effect of DDR-selective inhibitors in our assay system.
Collaborator Contribution Dr. Nathanael Gray provides us DDR-selective inhibitors.
Impact We obtained positive data and submitted programme grant to Arthritis Research UK.
Start Year 2014
 
Description Dr. Richard Williams 
Organisation University of Oxford
Department Kennedy Institute of Rheumatology
Country United Kingdom 
Sector Academic/University 
PI Contribution This is a direct collaboration on collagen-induced arthritis mouse model. We test the role of MT1-MMP in arthritis development.
Collaborator Contribution Dr. Williams provides us his expertise on the mouse model of arthritis.
Impact Paper is in preparation
Start Year 2009
 
Description Dyax 
Organisation Dyax
Country United States 
Sector Private 
PI Contribution Dyax provided us DX-2400 to test it on collagen induced arthritis model, and discovered it is effective.
Collaborator Contribution Dyax provided DX-2400
Impact As it is, it is still active, but getting uncertain as the company has changed their direction. They layed off the team I was working with.
Start Year 2009
 
Description Kadmon 
Organisation Kadmon Corporation
Country United States 
Sector Private 
PI Contribution Kadmon made exclusive deal with Dyax to develop DX-2400, MT1-MMP highly selective antibody inhibitor. As Dyax has been supplying DX-2400, I initiated forming a contract with Kadmon. It is almost end stage to finalise MTA.
Collaborator Contribution Kadmon provides us DX-2400 for our research now.
Impact Kadmon has just sent us 100 mg of DX-2400 they inherited from Dyax.
Start Year 2014
 
Description Kenneth Alrø Bøtkjær 
Organisation Aarhus University
Country Denmark 
Sector Academic/University 
PI Contribution Kenneth provides us antibody based highly selective MT1-MMP inhibitors. I was one of the collaborators to develop these inhibitors.
Collaborator Contribution He has provided us two inhibitors for cell culture studies.
Impact Kenneth has screened MT1-MMP inhibitors using phage display antibody library in University of Cambridge. He successfully obtained two strong candidates and optimised. They are single chain Fv-Fc fusion protein, and have very high affinity with high selectivity. He is now scaling up the production of these inhibitors to use for animal studies and cell culture models.
Start Year 2012
 
Description MT1-MMP inhibitor development 
Organisation Bicycle Therapeutics
Country United Kingdom 
Sector Private 
PI Contribution We are seeking to develop selective MT1-MMP inhibitors and made a collaboration with Bicycle therapeutics to develop such inhibitors. Our role is to screen the inibitors.
Collaborator Contribution They have developed candidate conpounds and we are now testing.
Impact we have now some lead and backup compounds that binds to hemopexin domain. They are now screening to identify the one which binds to catalytic domain. We are now in process of testing the compounds in our biological assays.
Start Year 2012
 
Description MT1-MMP inhibitory antibody/Prof Gillian Murphy 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution We examine the efficacy of MT1-MMP inhibitory antibodies for mouse model of arthritis as well as in vitro cell based assay systems.
Collaborator Contribution Professor Gillian Murphy at university of Cambridge provides us highly selective MT1-MMP inhibitory antibodies.
Impact This collaboration resulted MRC grant award in 2009.
Start Year 2009
 
Description Small molecule MMP/ADAM inhibitors 
Organisation University of Pisa
Country Italy 
Sector Academic/University 
PI Contribution We test small molecule MMP inhibitor in mouse model of arthritis and in vitro assays.
Collaborator Contribution Prof. Armando Rossello provides us small molecule MMP inhibitors.
Impact This collaboration resulted award of ERASMUS grant to the PhD student in the University of PISA.
Start Year 2015
 
Title DX-2400 
Description Dyax made DX-2400 which is a selective MT1-MMP inhibitory antibody. We collaborated with Dyax and tested efficacy of DX-2400 in collagen induced arthritis model. As our original antooibody inhibitor development was much delayed, we employed DX-2400 to carry our our projects. We discovered DX-2400 to suppress arthritis development. Especially it has significant effect when it si co-administrated with anti- TNF blockade. 
Type Therapeutic Intervention - Drug
Current Stage Of Development Refinement. Non-clinical
Year Development Stage Completed 2014
Development Status On hold
Impact Dyax licensed DX-2400 to Kadon, and I am now dealing with Kadmon to test DX-2400 further. 
 
Title MT1-MMP inhibitors 
Description We are still screening the compounds in collaboration with Bicycle therapeutics. 
Type Therapeutic Intervention - Drug
Current Stage Of Development Initial development
Year Development Stage Completed 2014
Development Status Under active development/distribution
Impact It is still not known yet. 
 
Description A session Chair for Metalloproteinase Gordon Research Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I chaired one of the sessions in the metalloproteinase Gordon Research Conference. The session was very active and many stimulated discussions.
Year(s) Of Engagement Activity 2019
URL https://www.grc.org/metalloproteases-conference/2019/
 
Description An Invited speaker for Matrix Biology Europe meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I was invited speaker in one of the session. I presented our recent study on role of MT1-MMP in rheumatoid arthritis in front of around 100 people who are international academic scientists and graduate students. I received several questions in discussion and after the session as well.
Year(s) Of Engagement Activity 2016
URL http://www.mbe2016.upatras.gr
 
Description An invited Keynote speaker for ShRImP Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact approximately 30 people attended from Germany and Italy, who are academics, medical doctors and post graduate students. Upon talk was given, active discussion took place. With this, I have engaged collaborations with German group from TUM.
Year(s) Of Engagement Activity 2015
 
Description An invited speaker for 47th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact There were over 100 audiences listening to my talk. The discussion was very active and many expressed their interests on my talk.
Year(s) Of Engagement Activity 2018
URL http://www.sbbq.org.br/reuniao/2019/images/livreto_2018_internet.pdf
 
Description An invited speaker for International Symposium of Matrix Biology 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The talk provoked active discussion.

It may result in new collaborative project.
Year(s) Of Engagement Activity 2015
 
Description An invited speaker for MMP Gordon Research Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I took a role of session chair and gave a talk as an invited speaker.around 95 scientists from all over the world have attended the meeting. My presentation provoked active discussion in the meeting, and I was appraised by the audience.
Year(s) Of Engagement Activity 2015
URL https://www.grc.org/programs.aspx?id=12358
 
Description An invited speaker for Symposium for "Future of Extracellular Matrix Research" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact There were over 80 scientists from Japan as audiences. The symposium was dedicated to Dr. Akira Ito's retirement.
"Future of Extracellular Matrix Research" (Tokyo, Japan, 22 March 2014)
Title of the talk: MT1-MMP: A Molecular Target in Rheumatoid Arthritis


I received over 10 positive questions/comments from audiences.
Year(s) Of Engagement Activity 2014
 
Description Euroscicon meeting 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Discussion workshop: Live Cell Imaging 17/06/2011
EuroSciCon
Title: Imaging Cellular Invasion: Role of the Cell Surface Collagen Degrading Enzyme
Author: Yoshifumi Itoh
I have discussed how we perform cellular invasion.

There are about 50 people attended.

Many discussions were made how we perform our invasion analyses. There are PhD students and Postdocs from achademics and some from bio-tech companies.
Year(s) Of Engagement Activity 2012
 
Description Invited as a speaker for MMP Gordon research conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I was one of the invited speakers in the MMP Gordon Research Conference. My talk was 15 min and there were active discussions and many question.
Year(s) Of Engagement Activity 2017
URL https://www.grc.org/matrix-metalloproteinases-conference/2017/
 
Description Invited as a speaker for Nordic Proteoglycan meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact around 50 international people attended to this meeting. I was one of the invited speakers and was given 3the longest slot for my talk. It triggered discussion and many questions were asked during and after the discussions.
Year(s) Of Engagement Activity 2018
URL http://norheart.no/nordic-proteoglycan-meeting-2018/
 
Description Invited keynote lecture for annual meeting of Japan Connective Tissue Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact My talk provoked active discussion.

The talk disseminated our findings to academic researchers working in the field of Matrix Biology in Japan.
Year(s) Of Engagement Activity 2014
URL http://www.jsctr.org/science_rally.htm
 
Description Invited speaker at Proteoglycan 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Over 150 attendees from different countries attended. I have given a 25 min seminar. I had good discussions.

I received good responses from different people.
Year(s) Of Engagement Activity 2013
URL http://www.proteoglycans2013.com
 
Description Invited speaker for FEBS Advanced Lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact I gave two lectures in this instance. In both lectures, I received many questions with interests during discussion times allocated and after lectures such as during lunch/dinner times.
Year(s) Of Engagement Activity 2017
URL http://www.febs-mpst2017.upatras.gr
 
Description Invited speaker of XVII meeting of the Brazilian Society for Cell Biology 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact I received several questions from audiences, and had a many other questions afterwards.

Many people gave me a very positive feed backs and had many questions afterwards with potential collaborations.
Year(s) Of Engagement Activity 2014
URL http://www.interevent.com.br/evento/sbbc2014
 
Description Invited speaker to 46th Sandbjerg Meeting on Membrane Transport 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Schools
Results and Impact 46th Sandbjerg Meeting on Membrane Transport
Date: Monday May 12th, 1.00 pm - Wednesday, May 14th, 2.00 pm 2014
Place Sandbjerg Estate, Sønderborg, Denmark
Topics:
1. Regulation and roles of proteases at the plasma membrane I
2. Regulation and roles of proteases at the plasma membrane II
3. How lipids cross cellular membranes
4. Free communications in membrane transport
My talk of title was: Spatiotemporal regulation of MT1-MMP determines cellular invasion.

Attending this meeting, I am initiating two collaborations with University of Copenhagen. The response of the talk by the audience was excellent.
Year(s) Of Engagement Activity 2014
URL http://membranes.au.dk/sandbjerg-meeting/
 
Description MMP Gordon Research conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other audiences
Results and Impact MMP Gordon Research Conference (Rhode Island, USA, August 2011)
Title: Invadopodia or not invadopodia? MT1-MMP localization to the substrate attachment site regulates cellular invasion.
Authors: Yoshifumi Itoh
I have presented our recent data how MT1-MMP is localized at invasion front of invading cells.

There aremany discussions made on this topics.
Year(s) Of Engagement Activity 2011
 
Description MMP gordon research conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Poster Presentation
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact 170 academics have attended the meeting.

The poster was selected as an oral presentation.
Year(s) Of Engagement Activity 2013
 
Description Meeting presentation at Winterschool of proteolytic enzymes and their inhibitors 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Paper Presentation
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Winter school of Proteiolytic enzymes and their inhibitors
Title: MT1-MMP activity is modified by other MT-MMPs
Authors: Kazuyo Kaneko and Yoshifumi Itoh

About 100 PhD and MSc students attended.


Many questions were raised.
Year(s) Of Engagement Activity 2012
 
Description Vice chancellors meeting at Oxford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact All high gear University of Oxford achademics have presented in this conference.
"Vice Chancellor's Symposium: Inflammatory mechanisms and inflammatory diseases"
Title: Cell trafficking: Role of cell surface metalloproteinase, MT1-MMP
Authors: Yoshifumi Itoh

I received may feed back indicating their interests to the works.
Year(s) Of Engagement Activity 2011