MULTIPLE SCLEROSIS 

In vitro

  • HTS / HCS Assays for the discovery of compounds inducing the differentiation of oligodendrocyte precursor cells (OPCs from mouse and human iPSCs expressing double fluorescence reporters)
  • In vitro system of myelinating co-culture : oligodendrocytes and neuronal (primary cells), brain slices, etc.

Read-out
Differentiation of OPCs in mature oligodendrocytes (switch of green to red fluorescent reporter)
Detection and monitoring of the myelin by quantitative detection of a specific (GFP or B-galactosidase) reporter gene system (transgenic mouse model PLP-GFP, Cnpase-GFP or MBP-LacZ)

Xenopus

  • Live-imaging model of the demyelination/remyelination process
Read-out
Quantification of oligodendrocytes expressing GFP along the optic nerve (transgenic tadpole MBP GFP-NTR), evaluation of efficiency of remyelination through a visual avoidance behavioral test

Rodents

Chemical demyelinating models
  • Lysolecithin local injections
  • Cuprizone diet
MOG-induced focal EAE: model of diffused inflammatory lesions in the spinal cord
MOG-induced EAE: model of focal lesions with inflammatory demyelination and remyelination in brain or spinal cord
Dysmyelinated mutants
Shiverer, PLP overexpression
Read-out
PET imaging (neuroinflammation, myelin, cerebral metabolic rate of glucose), longitudinal study using MRI (11.7T), histology, motor and cognitive behavior, etc.

Non-human primates

  • Model of lysolecithin-induced chemical demyelinisation of the optical nerve
  • EAE model
Read-out
PET imaging (neuroinflammation, myelin, cerebral metabolic rate of glucose), MRI, histology, motor and cognitive behavior, evoked visual potential, hematology, CSF cytokine measurement, etc.
Medium and High throughput genotyping, Histological asessment

Humans

  • Clinical investigation Center (Phase I and II), biological resource center, stratified cohorts of patients, genetic characterization of MS, analyses on tissus issued from MS patients
Read-out
PET and MRI imaging (demyelination/ remyelination, neuroinflammation, neuronal loss),
Medium and High throughput genotyping, Histological asessment
A comprehensive overview of our offer in the field of Multiple Sclerosis :
  • Basic Research

    Cellular models

    Cellular models for high throughput screen of molecules inducing the differentiation of oligodendrocyte precursor (OPC) cells (cell line CG4 with double fluorescence)

    - High throughput scree of molecules inducing the differentiation of oligodendrocyte precursor cells (OPCs) in mature oligodendrocytes on the basis of the expression of specific fluorescent reporter genes

    - Analysis of the intrinsic capabilities of putative candidates molecules inducing human OPCs differentiation (derived from human neural precursors and/or iPS cells

     

    In vitro system of myelinating co-culture : oligodendrocytes and neuronal (primary cells)

    - Detection and quantitative monitoring of myelin production by myelinating oligodendrocytes by assay of the enzymatic activity of a specific reporter gene (transgenic mouse model MBP-LacZ)

    - Detection and monitoring of the myelin by quantitative detection of a specific (GFP) reporter gene system (transgenic mouse model PLP-GFP)

    - Identification of hemato-encephalic barrier neuroprotective agents

    - Identification of permeabilizing agents of the hematoencephalic barrier and of the blood to CSF barrier

    References :

    - Stankoff B., Multiple sclerosis 1996 : vol. 2 p.125-132

    - Demerens, Neurology 1999 : vol.52 p. 346-350

    - Spassky et al. (2001) Dev. Neurosci. 23:318-326

    - Buchet et al. 2011, Brain134, 1168-1183

     

    Animal Models

    The xenopuse Model: Medium through-put screening of molecules favoring myelin repair by live-imaging the remyelinating process.

    - Quantification of oligodendrocytes expressing GFP along the optic nerve (transgenic tadpole pMBP-eGFP-NTR) in collaboration with Watchfrog (Abdelkrim Mannioui)

    References : Kaya F., Journal of Neuroscience 2012 : vol.32 p. 12885-12895

    - Evaluation of efficiency of remyelination through a visual avoidance behavioral test (under development).

     

    Murine models

    Chemical demyelinating Models

    - Lysolecithine local injections (corpus callosum and spinal cord tracts)

    - Cuprizone chronic diet inducing preferential lesions in the corpus callosumx

     

    Model of diffuse Experimental autoimmune encephalomyelitis (EAE MOG)

    - Leading to diffuse inflammatory lesions in the spinal cord.

    - Therapeutic efficacy of potential candidate molecules can be tested through the use of clinical rating scores correlated with histological analyses

     

    Model of focal Experimental autoimmune encephalomyelitis (MOG EAE-focal)

    - The model induces focal lesions with demyelination and inflammatory reaction to study the remyelination process

    - Consists in a pre-injection of a pro-inflammatory agent (IFNγ) followed by a TNFα injection into the corpus callosum or the dorsal tract of the spinal cord (Anne Baron)

     

    Dysmyelinate mutants

    Shiverer, PLP (PLOA) to screenmolecules in the context of intracerebral transplantation.

    Model of Experimental autoimmune encephalomyelitis through transfert of encephalitogenic lymphocytes of various polarities inducing spinal or cerebral demyelinating lesions

    References :

    - Kerschensteiner Martin, American Journal of Pathology 2004: vol. 164 p. 1455-1469

    - Tepavcevic Vanja, Journal of Clinical Investigation 2011: vol. 121 p. 4722-4734

    - Blanchard et al. J Neurosci. 2013 Jul 10;33(28):11633-42

    - El Behi Mohamed, Nat. Immunol. 2011 Jun;12(6) :568-75

    - El Behi Mohamed, J Neuroimmune Pharmacol. 2010 Jun;5(2) : 189-97

     

    Biobanks

    Biological ressource center CRB-REFGENSEP

    DNAs and PBMCs from 2700 clinically annotated patients and apparented (age of disease onset, form of disease, EDSS, treatments) et 700 healthy controls assessed for all 110 genetic factors known to predispose to the disease (available at BioCollections)

    (Bertrand Fontaine, isabelle Rebeix, Léna Guillot-Noël)

     

    Analyzing tools

    Histological analyses

    Immunohistochemistry

    Electron Microscopy

     

    Imaging tools

    - In vivo imaging :

    * micro-PET (Myelin sheath)

    * 2-photon microscopy (live imaging)

    * MRI 11.7T & 7T : T2-weighted imaging, DTI, localised proton spectroscopy, quantification through LC Model, glutamate CEST-imaging, Diffusion NMR spectroscopy of differents NMR detectable metabolites (NAA, Glu, Gln…etc)

    - Ex vivo Imaging : histological analyses

  • Preclinical Research
  • Clinical Research