A functional proteomic screen of myelination

Lead Research Organisation: University of Edinburgh
Department Name: MRC Centre for Regenerative Medicine

Abstract

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Technical Summary

Myelination represents one of the most spectacular cell-cell interactions in biology. Despite
this, we know very few of the signalling molecules involved in axo-glial adhesion and/or in
ensuring that the number of wraps is precisely related to axon diameter. Identification of
these molecules will provide therapeutic targets for remyelination strategies in Multiple
Sclerosis (MS), the failure of which results in the axon loss and progressive disability that
characterises the disease. As well as representing a major disease burden to sufferers and
their families, the chronic progressive nature of MS causes a significant cost to society –
currently estimated at 5-13 billion euros/annum in the EU. Currently there are no therapies
in clinical use or even in trials to promote remyelination, in large part due to the lack of
targets. We propose a novel approach to the problem by using a combination of
monoclonal antibody and recombinant phage display technology with FALI (fluorophoreassisted
light inactivation), a technique to create acute protein knockdown of antibodybinding
targets by singlet oxygen-mediated protein damage when the fluorophore is
exposed to light. We will generate libraries of monoclonal antibodies or single-chain
variable fragments (scFvs) by immunizing mice with oligodendrocytes, the myelin-forming
cells of the CNS. We will screen these for surface binding to oligodendrocytes, then couple
those that bind to a fluorochrome (FITC) and determine which inhibit myelination following
exposure to 490nm light in a neurone/oligodendrocyte myelinating co-culture assay.
Proteomics will then be used to identify the target antigens for those that inhibit
myelination, so defining signalling molecules for validation by conventional transgenic
techniques. The use of monoclonal antibody technology and FALI provides an unbiased
approach for the identification of key signalling molecules quite distinct from conventional
candidate-driven approaches.t therefore provides a method both to understand this
essential part of CNS development and also to identify entirely new targets for
remyelination therapies that will not be detected by conventionally-funded incremental
research programmes.

Publications

10 25 50
 
Description BIRAX
Geographic Reach Europe 
Policy Influence Type Membership of a guideline committee
 
Description Bristol review
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
 
Description 089000/Z/09/Z Wellcome Trust Programme Grant (Axoglial interactions during myelination in the CNS) PIs Brophy and French-Constant
Amount £1,610,619 (GBP)
Funding ID 089000/Z/09/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 08/2008 
End 07/2013
 
Description Are endothelin Receptors suitable targets for remyelination therapies?
Amount £100,706 (GBP)
Funding ID 950 
Organisation Multiple Sclerosis Society 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2012 
End 01/2015
 
Description Canadian MS Society posdoc fellowship
Amount $80,000 (CAD)
Organisation Multiple Sclerosis Society of Canada 
Sector Charity/Non Profit
Country Canada
Start 09/2017 
End 08/2020
 
Description EMBO
Amount £120,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Learned Society
Country European Union (EU)
Start 01/2012 
End 01/2015
 
Description MS Society Research Grants
Amount £242,367 (GBP)
Organisation Multiple Sclerosis Society 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2018 
End 12/2020
 
Description Marie Curie Fellowship
Amount € 195,000 (EUR)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 05/2018 
End 04/2020
 
Description University of Edinburgh - MS Society Clinical Science Centre
Amount £35,000 (GBP)
Organisation Multiple Sclerosis Society 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2011 
End 08/2014
 
Description Wellcome Trust Investigator award
Amount £1,900,000 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2015 
End 02/2020
 
Title CFFC - co-culture protocol that enables myelination signals to be assayed 
Description co-culture protocol that enables myelination signals to be assayed 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2012 
Provided To Others? Yes  
Impact By training post docs and students from labs in Europe in its use (2 in 2012). Publications by other groups and refinement of myelination assays by replacing in vivo assays with an in vitro model 
 
Title CffC Fibre cultures 
Description Nanofibres to assess level of oligodendrocyte myelination 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? Yes  
Impact Novel assay to look at myelination - used by other research groups 
 
Title FALI 
Description Use of fluorescent light to inactivate molecules on the surface of cells based on the release of free radicals by fluorochrome coupled antibodies attached to these cells 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Rapid assay for cell adhesion between axons and glai 
 
Title myelinating co-cultures 
Description A robust myelination assay using purified oligodendrocyte precursors and dorsal root ganglion neurones. Enables drugs etc to be rapidly screened 
Type Of Material Technology assay or reagent 
Year Produced 2009 
Provided To Others? Yes  
Impact Nature neuroscience paper from collaboration with Robin Franklin (Cambridge) using the assay to examine RXR signalling. 
 
Description Axoglial interactions 
Organisation University of Edinburgh
Department Centre for Neuroregeneration (CNR)
Country United Kingdom 
Sector Academic/University 
PI Contribution Analysis of cell adhesion molecules that regulate axoglial contact
Collaborator Contribution Intellectual input and regular meetings
Impact Wellcome Trust Programme Grant Publication 19451276
Start Year 2007
 
Description Braod image analysis 
Organisation Broad Institute
Country United States 
Sector Charity/Non Profit 
PI Contribution Definition of the problem of quantifying myelin sheaths
Collaborator Contribution Development of machine learning image analysi protocols
Impact none to date
Start Year 2017
 
Description Disc1 SC 
Organisation University of Edinburgh
Country United Kingdom 
Sector Academic/University 
PI Contribution New research tools - fibres
Collaborator Contribution Human cells and initial results
Impact none to date
Start Year 2017
 
Description Roche Knussel 
Organisation Roche Pharmaceuticals
Department Neurosciences
Country Global 
Sector Private 
PI Contribution Provision of expertise in fibre cultures
Collaborator Contribution Advice on upscaling to large scale screens for remyelination targets
Impact none so far
Start Year 2018
 
Description Septin Werner 
Organisation Max Planck Society
Department Max Planck Institute for Experimental Medicine
Country Germany 
Sector Public 
PI Contribution Discovery of myelin abnormalities that might be explained by septin mislocalization
Collaborator Contribution expert advice and septin antibodies
Impact none to date
Start Year 2017
 
Description Zebrafish collaboration 
Organisation University of Edinburgh
Department Centre for Neuroregeneration (CNR)
Country United Kingdom 
Sector Academic/University 
PI Contribution expertise in oligodendrocyte morphology during myelination
Collaborator Contribution Publication in Development
Impact Publication in development
Start Year 2010
 
Description joint projects on CNS remyelination 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution Post docs in my lab working on macrophage biology
Collaborator Contribution Specialist knowledge and experimental work on parabiosis model
Impact Co-publications since 2011 23026979 and 21904791 and 21469916 and 21131950
 
Description work on myelination and bilirubin 
Organisation Technical University of Lisbon
Country Portugal 
Sector Academic/University 
PI Contribution Trained student in her lab in cell culture
Collaborator Contribution Student working on project
Impact Co-publications since 2011 23086523
Start Year 2011
 
Description work on polarity 
Organisation University of Edinburgh
Country United Kingdom 
Sector Academic/University 
PI Contribution Post doc in my lab working on Scribble function in myelination
Collaborator Contribution Specialist knowledge and analysis of tissue samples
Impact No outputs in 2012
Start Year 2007
 
Title Patent - TECH ID TEC3348 - Charles ffrench-Constant 
Description BQ - Identification of Activin A as a novel target for remyelination therapy 
IP Reference WO2007083133 
Protection Patent granted
Year Protection Granted 2013
Licensed Commercial In Confidence
Impact Priority
 
Description Charles frrench-Constant lecture at MS Space North annual meeting 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact MS Space North annual meeting, 16 August 2013, Exhibition Centre, Aberdeen, Scotland. Prof Charles ffrench-Constant was invited to give a keynote lecture. 150 patients with MS and their families attended. Lecture was recorded and is available online (214 views so far): http://www.mssociety.org.uk/ms-events/2013/06/ms-space-north-2013-aberdeen.

Part of a general objective of disseminating information concerning regenerative medicine, generating interest in this field.
Year(s) Of Engagement Activity 2013
URL http://www.mssociety.org.uk/ms-events/2013/06/ms-space-north-2013-aberdeen
 
Description Edinburgh science festival 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Questions afterwards

none
Year(s) Of Engagement Activity 2014
 
Description RSE lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Health professionals
Results and Impact Discussion about MS therapies

None
Year(s) Of Engagement Activity 2014
 
Description Talks to MS Society meetings 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Primary Audience Participants in your research and patient groups
Results and Impact Platform presentations at MS Meetings for patients

Dissemination of information, and funding donations to Society
Year(s) Of Engagement Activity 2008,2009