Cellular models
In-vitro cell culture (primary cell cultures) of (mouse/rat) cortical and/or striatal neurons.
- Evaluation (through quantification of neuronal survival) of the properties of new molecules in a cell death model induced by different neurotoxins (quinolinate, glutamate, 3-nitropropionate), leading to oxidative stress, mitochondrial impairment and excitotoxicity.
- Evaluation (through quantification of neuronal survival) of the effects of new molecules in a cell death model induced by injection lentiviral vectors encoding different parts of the mutated huntingtin gene.
- Evaluation (through quantification of neuron survival) of the properties of new molecules in a spontaneous cell death model obtained from primary cell cultures (neurons/astrocytes) arising from transgenic mouse models of Huntington’s disease
In-vitro model of human ESC and/or iPS cells obtained from HD patients
- In vitro differentiation of cell lines issued from patients bearing different CAG repeats and from healthy controls
- Identification of cell death events or mechanisms of cellular dysfunction at play in these cells to identify potential new therapeutic targets
- Evaluation (through quantification of neuronal survival) of new drug- or gene-based strategies in this cell death model induced by different neurotoxins, ), leading to oxidative stress, mitochondrial impairment and apoptosis.
Animal models and associated analysis tools
Rodent models
In vivo rodent Models of striatal cell degeneration induced by neurotoxins
Excitotoxic striatal lesions in rat or mice induced by intrastriatal stereotactic injections of quinolinate (or otherglutamate agonists)
- Evaluation of the neuronal cell death, neurochemical deficits, motoric alterations, and neuroinflammation induced in these models,
- Evaluation of potential neuroprotective effects of drug- or gene-based therapeutic strategies on these models,
- Evaluation of potential neurorestorative strategies (cell-, gene- or drug-based) on motor symptoms in chronically lesioned animals
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Chronic systemic 3NP lesion model
- Longitudinal evaluation of potential neuroprotective strategies (either drug- or gene-based)
- Longitudinal evaluation of neurorestorative strategies (either cell-, gene- or drug-based) on motor symptoms in chronically lesioned animals
- follow-up of symptoms in chronically lesioned animals
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Genetic Models
Rodent (mouse/rat) models of progressive striatal degeneration induced by local injection of viral vectors (AAV/lentivirus) encoding various fragments of the mutated huntingtin gene
- Evaluation of neuroprotectrice strategies (drug-based, gene-based) through biochemical, transcriptomics and histological approaches
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Transgenic models of progressive degeneration and dysfunction in mouse (for example, the KI140CAG model)
- Evaluation of neuroprotectrice strategies (drug-based, gene-based) through biochemical, transcriptomics and histological approaches
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Lentiviral and/or AAV vectors to obtain proof of concept on potential therapeutic targets in different genetic models of Huntington’s disease
- Invalidation of target genes (ShRNA strategies) overexpression of recombinant proteins
Non-human Primate models
Neurotoxic lesion models
Excitotoxic striatal lesions in Macaca fascicularis induced by intrastriatal stereotactic injections of quinolinate (or other glutamate agonists)
- Evaluation of the neuronal cell death, neurochemical deficits, motoric alterations, and neuroinflammation induced in these models,
- Evaluation of potential neuroprotective effects of drug- or gene-based therapeutic strategies on these models,
- Evaluation of potential neurorestorative strategies (cell-, gene- or drug-based) on motor symptoms in chronically lesioned animals
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Chronic systemic 3NP lesion model in Macaca fascicularis
- Longitudinal evaluation of potential neuroprotective strategies (either drug- or gene-based)
- Longitudinal evaluation of neurorestorative strategies (either cell-, gene- or drug-based) on motor symptoms in chronically lesioned animals
- follow-up of symptoms in chronically lesioned animals
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Genetic Models
Models of progressive striatal degeneration induced by local injection of viral vectors (AAV/lentivirus) encoding various fragments of the mutated huntingtin gene
- Evaluation of neuroprotectrice strategies (drug-based, gene-based) through biochemical, transcriptomics and histological approaches
- Development of new imaging modalities for the in vivo monitoring of various disease processes (neuro-inflammation, cell death markers, etc...)
Lentiviral and/or AAV vectors to obtain proof of concept on potential therapeutic targets in different genetic models of Huntington’s disease
Invalidation of target genes (ShRNA strategies) overexpression of recombinant proteins
Preclinical materials and methods available
Biochemistry and Proteomics
- « Western blot » Quantitative Analyses
- ELISA
Transcriptomics
- MicroArray
- RNA seq
Microscopy and histological assessments
- Real time video-microscopy
- Automated epi-fluorescence microscopy
- Confocal Microscopy
- Neuropathological evaluation of lesions by a certified anatomo-pathologist (European board bearer)
- Immunohistochemical detection / immunofluorescence microscopy
- Unbiased stereological cell counts, morphometric analyses, 3D-reconstruction of histological serial section stacks
- High through-output multiresolution computer assisted virtual microscope (fluorescence and bright-field)
In vivo evaluation of lesions and therapeutic efficacy
Behavioural analyses
Rodents: :
- Video-based analysis of locomotor behavior (CatWalk, Rotarod) and cognitive evaluation (Morris water maze test, Novelty, Avoidance test, etc...)
Non-human primates: :
- Video-based analysis of locomotor activity (Ethovision, NOLDUS) and semi-quantitative clinical rating scale (The Observer, NOLDUS)
- Battery of cognitive tests using tactile screens on individual cages and/or gang band (social interactions)
* tests include the CANTAB battery of tests for non-human primates (DNMS, DMS, IEDS, CSST, Memory 30, Visual Discrimination Task, etc...)
* as well as build in batteries of tests developed for specific purposes
In vivo Imaging
- PET Imaging
* PET parametric imaging of the regional metabolic rate of glucose : 18F-FDG
* PET parametric imaging of the neuro-inflammation : 18F-DPA714
* Compartmental modeling and parametric imaging, automatized input function, PET/MRI co-registration (neuroFocus 220/ MRI 11.7T)
- Very high field MR imaging / MR spectroscopy of rodents (11.7 T Bruker for rodents, 7T Varian for NHP)
* T2-weighted imaging
* Water diffusion
* Proton NMR spectoscopy, quantification through LC Model
* Glutamate CEST-imaging
* Diffusion NMR Spectroscopy of various NMR detectable metabolites (NAA, Glutamate, Glutamine, etc…).
Post-mortem analyses (rodent & non human primates)
- Immunohistochemistry
- In situ Hybridation
- 3D Reconstruction of whole brain histological volumes (multimodal imaging)
- In vivo/post-mortem 3D co-registration of PET/MRI and 3D-histological volumes
- Biochemical detection of disease markers in various biological fluids (bllod, plasma, CSF, etc…)
- Reglementary anatomo-pathological studies (rodent/primate) by a trained anatomopathologist (European board bearer)