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P3‐430: Gene and cell therapy approaches for prion diseases
Author(s) -
Gabelle Audrey,
Gabelle Audrey,
RelanoGines Aroa,
Hamela Claire,
Perrier Véronique,
Belondrade Maxime,
Lin Y.L,
De Vidi Isabelle,
Mettling Clément,
Corbeau Pierre,
Lehmann Sylvain,
Crozet Carole
Publication year - 2011
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2011.05.1874
Subject(s) - biology , genetic enhancement , neural stem cell , caudate nucleus , mutant , cell , stem cell , hippocampus , microbiology and biotechnology , gene , neuroscience , genetics
Background: Prion diseases are fatal neurodegenerative disorders for which there is currently no effective treatment. They are characterized by neuronal loss, vacuolation, astrocyte proliferation and by the accumulation in the brain of a pathologic protein called PrP. The PrPisoform results from the PrPc host-encoded protein trans-conformation. Since the potential for late-stage therapies using chemical molecules seems limited, therapeutic cell replacement strategies have raised optimistic promises. The conceptual idea underlying stem cell graft approaches is to replace non-functional cells by healthy and resistant ones that could deliver anti-prionmolecules of interest. Our objective is to combine cell and gene therapy. Our strategy is to use genetically modified ES cells to make grafts inscrapie infected mouse brain. In order to orchestrate the brain repair, our transplanted ES cells were engaged in early neuronal differentiation stage and were genetically modified by introducing “dominant negative” PrP mutants (DN-PrP) usinglentiviral vectors. The DN-PrP cells were produced by taking advantage of prion “resistant” polymorphisms Q171R and E219K that naturally exist in sheep and humans, respectively. Methods: To determine the best ES neuronal differentiation protocols, we quantitatively and qualitatively compared three major protocols (Mc Kay, Smith, Benninger). These protocols lead to neural stem cell called ES-NSC. By using FIV derivedlentivirus, we succeeded in transducing the ES-NSC with lentivirus carrying the DN-PrP mutants. Stereotaxic grafts were performed in wild type and also in prioninfected C57bl/6 mice. Specific sites of injection were selected such as the hippocampus, the caudate nucleus, and the lateral ventricles. We compared the duration of the disease and the neuropath logical profil in grafted or nongrafted mice. Results: The best characterization of neuronal engaged ES cells is obtained with the Smith differentiation protocol when comparing with the Mc Kay and Benninger protocols. We are now assessing the effect of the transplantation on the development of the disease. Conclusions: Our preliminary results of this combined gene and stem cell therapy based on Dominant Negative effecting Prions diseases appear to be promising.