Community Research and Development Information Service - CORDIS

ERC

REPROPARK Report Summary

Project ID: 340527
Funded under: FP7-IDEAS-ERC
Country: Italy

Periodic Report Summary 3 - REPROPARK (New experimental therapeutic approaches for Parkinson’s disease by direct DA neuronal reprogramming)

The overarching goal of this project is to optimize strategies for direct reprogramming of adult skin fibroblasts into functional and transplantable induced dopaminergic (iDA) neurons for establishing a cell therapy approach for Parkinson’s disease (PD). In fact, although mouse cells can be reprogrammed to iDA neurons with high efficiency, the same approach is much less effective in adult human cells. For this, we have implemented new reprogramming strategies combining the activation of genes together with a combination of small molecules able to reprogram adult human fibroblasts into iDA neurons with a robust efficiency up to 70%. The human iDA neurons acquired multiple functional properties including excitable membrane, sustained firing profile, synaptic competence and dopamine production. This reprogramming strategy has been, then, successfully applied to generate iDA neurons from monkey and porcine skin fibroblasts. Next, we asked whether this protocol could be applied to other adult human cell types with higher translational potential. For this, mesenchymal stem cells were isolated from the adipose tissue of adult donors and subjected to cell reprogramming. Interestingly, this procedure enabled the reprogramming of this cell type generating iDA neurons with mature properties with a significant efficiency. These results indicate that this reprogramming method is efficient in generating iDA neurons starting from different adult cell populations from both humans and large animals. Finally, we aimed to increase the safety of this approach enhancing its translational potential by eliminating the use of gene integrating lentiviruses. For this aim, we cloned a multicistronic cassette expressing the reprogramming transgene in an AAV vector. Then, AAV particles were produced and used to reprogram adult human fibroblasts in combination with the small molecule cocktail. Remarkably, this method led to a substantial generation of fully functional iDA neurons without any genomic trace of the reprogramming event. These studies establish a novel procedure for the safe and effective reprogramming of adult human cells into iDA neurons generating a very promising cell source for therapeutic approaches of cell therapy in brain disorders.

Reported by

Ospedale San Raffaele
Italy
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