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Theodorou, M. ; Rauser, B. ; Zhang, J. ; Prakash, N. ; Wurst, W. ; Schick, J.

Limitations of in vivo reprogramming to dopaminergic neurons via a tricistronic strategy.

Hum. Gene Ther. Methods 26, 107-122 (2015)
DOI PMC
Parkinson's Disease is one of the most common neurodegenerative disorders characterized by cell death of dopaminergic neurons in the substantia nigra. Recent research has focused on cellular replacement through lineage reprogramming as a potential therapeutic strategy. This study sought to use genetics to define somatic cell types in vivo amenable to reprogramming. To stimulate in vivo reprogramming to dopaminergic neurons, we generated a Rosa26 knock-in mouse line conditionally overexpressing Mash1, Lmx1a and Nurr1. These proteins are characterized by their role in neuronal commitment and development of midbrain dopaminergic neurons and have previously been shown to convert fibroblasts to dopaminergic neurons in vitro. We show that a tricistronic construct containing these transcription factors can reprogram astrocytes and fibroblasts in vitro. However, cassette overexpression triggered cell death in vivo, in part through endoplasmic reticulum stress, while we also detected uncleaved forms of the polyprotein, suggesting poor cleavage efficiency of the 2A peptides. Based on our results, the cassette overexpression induced apoptosis and precluded reprogramming in our mouse model. Therefore, we suggest that alternatives must be explored to balance construct design with efficacious reprogramming. It is evident that there are still biological obstacles to overcome for in vivo reprogramming to dopaminergic neurons.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Unfolded Protein Response; Embryonic Stem-cells; Human Fibroblasts; Dna Methylation; Gene-expression; Mouse Fibroblasts; Defined Factors; Er Stress; Functional-neurons; Transgenic Mice
ISSN (print) / ISBN 1946-6536
e-ISSN 1946-6544
Zeitschrift Human Gene Therapy : Methods
Quellenangaben Band: 26, Heft: 4, Seiten: 107-122 Artikelnummer: , Supplement: ,
Verlag Mary Ann Liebert
Verlagsort New York, NY
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Developmental Genetics (IDG)