Isabelle de Luzy

Recipient of scholarship
Project:  Improving the safety and function of pluripotent stem cell-derived neural transplants for the treatment of Parkinson's disease.

Human pluripotent cells are a promising tool for ‘cellular replacement therapy’, whereby these cells can replace lost ventral dopaminergic neurons (vmDA) in Parkinson’s disease patients.
Despite the successful generation of bona fide vmDA neurons in vitro, the asynchronous, heterogenous nature of differentiation means in vivo graft populations consist of proliferative, immature and terminally differentiated cells. It is therefore imperative to remove these poorly specified and potentially tumorigenic cells.
I propose to use dual techniques to improve both the safety and function of pluripotent stem cell-derived neural transplants:
1) Utilise cell sorting strategies (FACS) to isolate dopaminergic progenitors. This will be achieved using hESC reporter lines (LMX1A-GFP and PITX3-GFP; to isolate early and late dopamine progenitors) as well as isolation of progenitors based upon expression of cell-surface antigens. Such approaches will standardize and enrich donor preparations for in vivo transplantation, as well as identify cell surface marker candidates to purse.
2) Selective ablation of human embryonic stem cell-derived subpopulations based upon the activation of genetically engineered suicide genes. We will make use of the Thymidine kinase/ganciclovir suicide gene system to generate a number of new human stem cell lines that can be used in transplantation, and for which suicide can be initiated in vivo of select populations of cells (such as dividing cells or non-dopaminergic neurons) following administration of the drug gangciclovi to overcome the highe degree of heterogeneity present in differentiated populations.
Both of these strategies will be trialed using differentiated human stem cell lines implanted into rat models of neurodegenerative disease.
3) Employ neuroprotective interventions to improve the survival of vmDA neurons. We will trial 4 anti-apoptotic agents and assess their impact on vmDA neuron survival after passaging in vitro and post-transplantation in vivo.
Project: Improving the safety and function of pluripotent stem cell-derived neural transplants for the treatment of Parkinson's disease.
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