Manufacturing hiPSCs for therapeutic applications in Parkinson’s Disease: a GPC4-targeting approach for efficient generation of "safe" midbrain dopaminergic neurons.
Human induced pluripotent stem cells (hiPSCs) from Parkinson’s disease (PD) patients are expected to have strong therapeutic impact for PD given their application in disease modelling, drug discovery and cell replacement therapy. There are however important constraints to overcome before these hiPSCs can be used at best. Besides showing low propensity to generate midbrain dopamine neurons (mDAn), the mDAn yields are highly variable among different lines, thus limiting development of highly standardised procedures for in vitro application and for clinics.
HomeNeurojobsManufacturing hiPSCs for therapeutic applications in Parkinson’s Disease: a GPC4-targeting approach for efficient generation of “safe” midbrain dopaminergic neurons.
During this internship the student research will address these unmet issues for PD management by focusing on the morphogen regulator GLYPICAN-4 (GPC4). The first goal is to establish an unprecedented strategy for improving mDAn generation from patient-derived hiPSCs. We have previously shown that down-regulation of GPC4 confers to hiPSCs a unique biological state named “safe-PSC state” characterised by enhances propensity for mDAn lineage entry and differentiation. With this project the student will investigate the differentiation properties of patient-derived hiPSCs with down-regulated GPC4 levels to provide “proof-of-principle” that this strategy is compatible with efficient, reproducible and homogeneous mDAn differentiation of hiPSC lines from different origins.
In view of the potential therapeutic application of hiPSCs in a “safe-PSC state”, we have generated GPC4-blocking agents that permit to interfere with GPC4 function without involving genetically manipulation. The second goal of this project is to use these GPC4-blocking agents to develop efficient mDAn differentiation protocols of potential clinical relevant impact.
Major methodologies: 1) hiPSCs cell culture, expansion, and differentiation; 2) RT-qPCR; 3) immunostaining; 4) loss-of-function studies; 5) fluorescence microscopy.