Limitations of In Vivo Reprogramming to Dopaminergic Neurons via a Tricistronic Strategy
| العنوان: | Limitations of In Vivo Reprogramming to Dopaminergic Neurons via a Tricistronic Strategy |
|---|---|
| المؤلفون: | Benedict Rauser, Jingzhong Zhang, Nilima Prakash, Marina Theodorou, Wolfgang Wurst, Joel A. Schick |
| المصدر: | Human gene therapy / Methods 26(4), 107-122 (2015). doi:10.1089/hgtb.2014.152 |
| بيانات النشر: | Mary Ann Liebert Inc, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | Ascl1 protein, mouse, Male, Somatic cell, Gene Expression, Applied Microbiology and Biotechnology, Lmx1a protein, mouse, Mice, Transduction, Genetic, Gene Order, Nuclear Receptor Subfamily 4, Group A, Member 2, Basic Helix-Loop-Helix Transcription Factors, Transgenes, Homologous Recombination, Genetics (clinical), Genetics, metabolism [Astrocytes], Cell Death, metabolism [Dopaminergic Neurons], Dopaminergic, genetics [Transcription Factors], Cellular Reprogramming, Cell biology, genetics [Cellular Reprogramming], genetics [Cell Death], Molecular Medicine, Female, Nr4a2 protein, mouse, Reprogramming, Programmed cell death, genetics [LIM-Homeodomain Proteins], Genetic Vectors, LIM-Homeodomain Proteins, genetics [Basic Helix-Loop-Helix Transcription Factors], Mice, Transgenic, Substantia nigra, Biology, genetics [Unfolded Protein Response], Cell Line, In vivo, Animals, ddc:610, Transcription factor, Embryonic Stem Cells, genetics [Lentivirus], cytology [Dopaminergic Neurons], Pharmacology, genetics [Nuclear Receptor Subfamily 4, Group A, Member 2], genetics [Genetic Vectors], Dopaminergic Neurons, Lentivirus, Fibroblasts, Embryonic stem cell, nervous system, Astrocytes, Unfolded Protein Response, Transcription Factors |
| الوصف: | 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. |
| تدمد: | 1946-6544 1946-6536 |
| DOI: | 10.1089/hgtb.2014.152 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::710c6b6b8dc78d56ee894acb76f9a916 https://doi.org/10.1089/hgtb.2014.152 |
| Rights: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....710c6b6b8dc78d56ee894acb76f9a916 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19466544 19466536 |
|---|---|
| DOI: | 10.1089/hgtb.2014.152 |