Neurodegenerative diseases

Team:

Anselme L. Perrier : Research Director DR2 (Inserm U861, Group Leader)

Simona Gribaudo: Post-doctoral Fellow (Inserm – ANR/ Labex « Revive »)

Morgane Louessard: Post-doctoral Fellow (Inserm – JPND « ModelpolyQ »)

Marie Michael: Associate engineer (CECS)

Axel Maulet: Associate engineer (CECS)

Julie Bigarreau: PhD Student UEVE (Inserm – ANR/ Labex « Revive »)

Célia Joseph: Master Student (UPMC)

Aims and background:

The group’s objectives are focused on using neural progenies of human Pluripotent Stem (hPS) cells to understand and develop new treatments for Huntington Disease (HD). This devastating neurodegenerative disorder belongs to a family of genetic diseases caused by mutations that expand a coding CAG repeat tract. The mean age of onset is 35, around 6,000 people are affected in France. No disease modifying therapies are available; HD therefore follows a slow progressive evolution leading to death within 15 years of onset of first symptoms. The team is conducting two major research programs dedicated to cell therapy on one hand and to pathological modeling and drug screening on the other hand.

 

Research programs:

Bench-to-bedside cell therapy program for HD

HD is partially amenable to treatment by substitutive cell therapy. However, this technique is marred by logistic problems that restrict considerably the number of patients who may benefit from it. A potent alternative source of cells is therefore acutely needed. Due to their original properties, hPS cells are prime candidates whose relevance has already been demonstrated for cell therapy of Parkinson’s disease. Our long term goal is to promote the clinical application of such cells for HD.

This translational cell therapy program is conducted within a European consortium (FP7 Repair-HD: http://www.repair-hd.eu). The rational is to use hPSCs as potent sources of cells to produce cell therapy product (CTP) for HD patient. We focus more specifically on: (A) GMP translations of CTP production and Quality Control assay development to evaluate such production (in collaboration with Roslin Cells, UK). (B) Pre-clinical in vivo functional testing in rodent and non-human primate models of HD (in collaboration with Dr. Hantraye & Dr R Aron-Badin CEA-MIRCen and Dr. Dunnett, Cardiff University, UK). (C) Immunogenicity of PSC-derived CTP: recipient response to allogeneic CTP in non-human primate models of HD (in collaboration with Dr Hantraye/Aron-Badin and Pr E Cozzi, Padova, Italy)

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Disease modelling with disease or patient-specific hPSC

Even though it may correct existing neuronal losses, cell replacement is not by itself a cure for HD as it cannot stop the progression of the neurodegeneration in the patient’s brain. In addition to substitutive therapy, efficient neuroprotective treatments should therefore be implemented. The search for such a complementary treatment is a major endeavour. Despite a wealth of existing cellular models, this undertaking probably requires additional cellular models of HD more closely replicating the patient’s situation. We postulate that HD mutant hPSC (embryonic or induced) lines can constitute such models and would be key to the exploration of the molecular mechanisms of the disease and ultimately allow the screening of therapeutic compounds endowed with a therapeutic potential.

Our current objectives is to find new insights in both cell autonomous and non-cell autonomous pathological cascades in HD. The rational is to make the most of the key properties of disease-specific hPSCs as versatile and highly relevant tools and biological substrates to study HD. We develop protocols, vector and lines to challenge HD genetic determinant in selected relevant cell populations (cortical or striatal neurons, astrocytes) alone or integrated within in vitro neuronal network in microfluidic chambers. Next we these tools to explore how mutations in the HTT protein impact on human cortical development and human neuronal and astroglial functions. Finally we aim at identifying phenotype specific transcriptional targets of mutant HTT (mut-HTT) or HTT loss of function in relevant neuronal/glial populations.

Drug discovery for HD with hPSC

Our general aim is to identify therapeutically relevant signaling pathways and lead-compound(s) for HD. We more precisely develop molecular and phenotypic screening assays to report HD-mediated functional impairments in HD-hPSC neuronal or glial. Next we use such assays to identify LEAD chemical compound(s) with therapeutic potential for HD or to validate drug or gene-based therapeutic approaches developed by our collaborators (Pr. Nicole Deglon, CHUV, Laussanne Swiss) or industrial partners.

 

 

Publications (here)

 

Patent:

WO/2010/063848 (2010-06-10)

BENCHOUA, Alexandra; (FR). PERRIER, Anselme; (FR). AUBRY, Laetitia; (FR).

Method and medium for neural differentiation of pluripotent cells

 

Collaborations:

 

 

MIRCEN – CEA (Fontenay-aux-Roses) – CEA CNRS UMR9199

Dr. Philippe Hantraye & Dr. Romina ARON-BADIN (FP7-Repair-HD)

Dr. Emmanuel Brouillet

 

CNRS UMR7102, UPMC Paris

Dr. Jean-Michel Peyrin (ERANet Neuron Microdeg)

 

CNRS UMR9197, Gif-sur-Yvette, France

Dr. Ronald Melki

 

CHUV, LABORATOIRE DES NEUROTHÉRAPIES CELLULAIRES ET MOLÉCULAIRES, Lausanne, (Swiss)

Dr. Nicole Deglon

 

Cardiff University, School of Biosciences

Steve Dunnett et Meng Li (FP7-Repair-HD)

 

Roslin Cells Limited, Edinburgh, UK

Dr. Catherine Jomary (FP7-Repair-HD)

 

Brain Repair and Imaging in Neural Systems, Lund University, Sweden

Dr. Deniz Kirk

Latest publication:

 

[Collaboration with Pr Deglon lab, CHUV, Laussanne, Swiss]

The Self-Inactivating KamiCas9 System for the Editing of CNS Disease Genes.

Merienne N, Vachey G, de Longprez L, Meunier C, Zimmer V, Perriard G, Canales M, Mathias A, Herrgott L, Beltraminelli T, Maulet A, Dequesne T, Pythoud C, Rey M, Pellerin L, Brouillet E, Perrier AL, du Pasquier R, Déglon N.

Cell Rep. 2017 Sep 19;20(12):2980-2991. doi: 10.1016/j.celrep.2017.08.075.

The KamiCas9 self-inactivating editing system allows transient expression of the Cas9 protein and high editing efficiency of  Huntington's disease (HD)

 

Preclinical Evaluation of a Lentiviral Vector for Huntingtin Silencing.

Cambon K, Zimmer V, Martineau S, Gaillard MC, Jarrige M, Bugi A, Miniarikova J, Rey M, Hassig R, Dufour N, Auregan G, Hantraye P, Perrier AL, Déglon N.

Mol Ther Methods Clin Dev. 2017 May 11;5:259-276. doi: 10.1016/j.omtm.2017.05.001. eCollection 2017 Jun 16.

PLoS One. 2016 Feb 10;11(2):e0148680. doi: 10.1371/journal.pone.0148680. eCollection 2016.

Dominant-Negative Effects of Adult-Onset Huntingtin Mutations Alter the Division of Human Embryonic Stem Cells-Derived Neural Cells.

Abstract

Mutations of the huntingtin protein (HTT) gene underlie both adult-onset and juvenile forms of Huntington's disease (HD). HTT modulates mitotic spindle orientation and cell fate in mouse cortical progenitors from the ventricular zone. Using human embryonic stem cells (hESC) characterized as carrying mutations associated with adult-onset disease during pre-implantation genetic diagnosis, we investigated the influence of human HTT and of an adult-onset HD mutation on mitotic spindle orientation in human neural stem cells (NSCs) derived from hESCs. The RNAi-mediated silencing of both HTT alleles in neural stem cells derived from hESCs disrupted spindle orientation and led to the mislocalization of dynein, the p150Glued subunit of dynactin and the large nuclear mitotic apparatus (NuMA) protein. We also investigated the effect of the adult-onset HD mutation on the role of HTT during spindle orientation in NSCs derived from HD-hESCs. By combining SNP-targeting allele-specific silencing and gain-of-function approaches, we showed that a 46-glutamine expansion in human HTT was sufficient for a dominant-negative effect on spindle orientation and changes in the distribution within the spindle pole and the cell cortex of dynein, p150Glued and NuMA in neural cells. Thus, neural derivatives of disease-specific human pluripotent stem cells constitute a relevant biological resource for exploring the impact of adult-onset HD mutations of the HTT gene on the division of neural progenitors, with potential applications in HD drug discovery targeting HTT-dynein-p150Glued complex interactions.