Edinburgh Research Explorer

Trf of Millstein award from Univ of Cambridge : A Functional Proteomic Screen of Myelination

Project: Research

StatusFinished
Effective start/end date1/09/0831/12/09
Total award£190,333.00
Funding organisationMRC
Funder project referenceG0601744/93956
Period1/09/0831/12/09

Description

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.

Research outputs