Functional Neuroprotection and Efficient Regulation of GDNF Using Destabilizing Domains in a Rodent Model of Parkinson’s Disease
| Autor: | Christina Isaksson, Erika Elgstrand Wettergren, Cecilia Lundberg, Giuseppe Manfré, Luis Quintino, Angrit Namislo |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Recombinant Fusion Proteins
animal diseases Genetic Vectors Neuroprotection Trimethoprim Rats Sprague-Dawley Neurotrophic factors Cell Line Tumor Drug Discovery Gene expression Escherichia coli Genetics Glial cell line-derived neurotrophic factor Animals Humans Glial Cell Line-Derived Neurotrophic Factor Molecular Biology Furin Neurons Pharmacology Regulation of gene expression biology urogenital system Escherichia coli Proteins Lentivirus HEK 293 cells Reproducibility of Results Parkinson Disease Genetic Therapy Molecular biology Rats Cell biology Disease Models Animal Tetrahydrofolate Dehydrogenase HEK293 Cells Neuroprotective Agents Gene Expression Regulation nervous system biology.protein Molecular Medicine Original Article Female GDNF family of ligands |
| Zdroj: | Molecular Therapy. 21:2169-2180 |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2013.169 |
| Popis: | Glial cell line-derived neurotrophic factor (GDNF) has great potential to treat Parkinson's disease (PD). However, constitutive expression of GDNF can over time lead to side effects. Therefore, it would be useful to regulate GDNF expression. Recently, a new gene inducible system using destabilizing domains (DD) from E. coli dihydrofolate reductase (DHFR) has been developed and characterized. The advantage of this novel DD is that it is regulated by trimethoprim (TMP), a well-characterized drug that crosses the blood-brain barrier and can therefore be used to regulate gene expression in the brain. We have adapted this system to regulate expression of GDNF. A C-terminal fusion of GDNF and a DD with an additional furin cleavage site was able to be efficiently regulated in vitro, properly processed and was able to bind to canonical GDNF receptors, inducing a signaling cascade response in target cells. In vivo characterization of the protein showed that it could be efficiently induced by TMP and it was only functional when gene expression was turned on. Further characterization in a rodent model of PD showed that the regulated GDNF protected neurons, improved motor behavior of animals and was efficiently regulated in a pathological setting. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|