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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

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

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

- Cuprizone chronic diet inducing preferential lesions in the corpus callosum

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 mutantsinic

- Dysmyelinic 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

Non-human Primate Models

Model of lysolecithin-induced chemical demyelinisation of the optical nerve (Anne Baron)

References :

- Bachelin et al., 2005 Brain 128 :540-9

- Lachapelle et al., 2005 J. Brain Pathol 15 :198-207


Analyzing tools : Measures of therapeutic efficacy of new agents

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

- Immunohistochemistry

- Electron Microscopy

Other analyzing tools

- Behavioural locomotor tests (rotarod, catwalk etc…)

- Clinical scoring

Clinical Research

Phase 1 and Phase 2 clinical trials

Clinical investigation Center of Pitié-Salpétrière

Genetic Characterization of MS. Search for genetic, transcriptomics and immunologic biomarkers of disease progression.

Medium and High throughoutput Genotyping

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 BioCollection)

Analyses on samples from MS patients ( in collaboration with the MS/NeuroCEB brain bank: Neurobiotech® )


Imaging Tools

Demyelinisation/remyelinisation by PET and MR imaging

Evaluation demyelination/remyelination through indirect (MRI) and direct (PET) imaging methods of analysis

Analyses on tissus issued from MS patients

Analysis of signaling pathways and/or therapeutic targets through histological asessment of MS tissus, in collaboration with the MS/NeuroCEB brain bank

Clinical PET imaging (on the SHFJ site)

Marker of the neuroinflammation : 18F-DPA714

In vivo evaluation of dysmyelinisation and remyelinisation :11C-PIB

Neuronal loss quantification : 11C-Flumazenil

References :

- Hammond et al. Neuron (2014)

- Huang et al. Nature Neuroscience (2011)

- Tepavcevic et al. (Annals Neurol, soumis)

- Stankoff et al. PNAS, Annals of Neurology

- Damotte et al. Genes Immun. 2014 Mar ;15(2) :126-32

- International Multiple Scleroris Genetics Consortium (IMSGC), Nat Genet. 2013 Nov. 45(11):1353-60