Functional identification of molecules promoting midbrain dopaminergic fate and neuritogenesis from embryonic stem cells

Lead Research Organisation: Imperial College London
Department Name: Institute of Clinical Sciences

Abstract

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Technical Summary

Parkinson‘s disease (PD) is a terminal progressive neurodegenerative disorder that affects 120,000 individuals in the UK alone. Most therapeutic strategies attempt to augment declining nigrostriatal dopamine function with drug or cell-based dopamine replacement strategies. However, the development of therapeutic paradigms via either approach is limited by a lack of sufficient understanding of the cellular and molecular mechanisms underlying differential vulnerability of substantia nigra dopaminergic (DA) neurons.

My objectives therefore are to understand the molecular make up of the midbrain DA lineage, to investigate the roles played by these molecules in midbrain development, and to apply these molecules in promoting DA fate determination and axon growth from stem cells. This knowledge would facilitate the development of stem cell based therapeutic strategies for treating PD and may provide insight into development of pharmaceutical drug targets. The experimental design is based on DA differentiation using embryonic stem cell model system and modern mouse genetic approach. We have generated ES cells and mice that express eGFP specifically in midbrain dopaminergic neurons via gene targeting in the midbrain DA specific Pitx3 gene. We have shown that these ES cells serve as a valuable cellular system to track midbrain DA differentiation and functional evaluation of potential DA regulators. We have carried out a microarray analysis comparing the expression profile of embryonic day 12 DA neurons with that of Pitx3-deficient DA neurons and midbrain non-DA cells, using FACS purified cell population. This study identified candidate transcription factors and axon guidance cues which provide the fundation for works proposed in this application.

The anticipated outcome of this application is the identification of key determinants for midbrain DA neurons and factors regulating neuritogenesis of these cells. From a therapeutic stand point, this work will identify nov target to push stem cells towards a dopaminergic fate in vitro which provides as a basis for their potential in developing pharmaceutical and cell based therapies for treating Parkinson‘s disease. Furthermore, the proposed study also allows further development of the Pitx3-GFP, Pitx3-taulacZ and Pitx3-/- ES cell systems and this in turn would facilitate the future exploitation of these technologies in Parkinson‘s related pharmaceutical drug screen program.

Publications

10 25 50
 
Description EC FP7 funding
Amount £500,000 (GBP)
Organisation European Commission 
Department Seventh Framework Programme (FP7)
Sector Public
Country European Union (EU)
Start 12/2008 
End 11/2012
 
Description Project Grant - Generation and characterisation of cortical interneurons from human pluripotent stem cells
Amount £171,841 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 10/2012 
End 09/2016
 
Title transgenic mouse line 
Description A mouse line that express a reporter in midbrain dopamine neurons and their axons 
Type Of Material Technology assay or reagent 
Year Produced 2009 
Provided To Others? Yes  
Impact N/A 
 
Description Wnt 
Organisation University of Edinburgh
Department Centre for Integrative Physiology
Country United Kingdom of Great Britain & Northern Ireland (UK) 
Sector Academic/University 
PI Contribution Wnt and neural differentiation
Collaborator Contribution Data and conceptual exchange
Impact Publication 18315501
 
Description Lab visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Primary Audience Public/other audiences
Results and Impact Talked about potential stem cell application to family members of Parkinson's disease patients

not known
Year(s) Of Engagement Activity 2008