title: Tight Long-term dynamic doxycycline responsive nigrostriatal GDNF using a single rAAV vector.
abstract: Glial cell line-derived neurotrophic factor (GDNF) gene transfer is being developed as a treatment for Parkinson's disease (PD). Due to the potential for side effects, external transgene regulation should enhance this strategy's safety profile. Here, we demonstrate dynamic control during long-term expression of GDNF using a recombinant adeno-associated virus (rAAV)-based bicistronic tetracycline (tet)-off construct. Nigrostriatal GDNF overexpression induces body weight alterations in rodents, enabling longitudinal in vivo tracking of GDNF expression after nigral vector delivery. Regulated GDNF expression was highly sensitive to dietary doxycycline (DOX), displaying undetectable striatal GDNF levels at serum DOX levels below those required for antimicrobial activity. However, in the absence of DOX, striatal GDNF levels exceeded levels required for efficacy in PD models. We also demonstrate the absence of a series of known GDNF-associated side effects when using direct intrastriatal vector delivery. Therefore, this single rAAV vector system meets most of the requirements for an experimental reagent for treatment of PD.
authors.affiliation: Department of Neuroscience, Powell Gene Therapy Center, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida 32611, USA.
authors.forename: Fredric P
authors.initials: FP
authors.lastname: Manfredsson
authors.forename: Corinna
authors.initials: C
authors.lastname: Burger
authors.forename: Aaron C
authors.initials: AC
authors.lastname: Rising
authors.forename: Kheir
authors.initials: K
authors.lastname: Zuobi-Hasona
authors.forename: Layla F
authors.initials: LF
authors.lastname: Sullivan
authors.forename: Alfred S
authors.initials: AS
authors.lastname: Lewin
authors.forename: Julia
authors.initials: J
authors.lastname: Huang
authors.forename: Emily
authors.initials: E
authors.lastname: Piercefield
authors.forename: Nicholas
authors.initials: N
authors.lastname: Muzyczka
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Mandel
mesh_terms: D000818:Animals; D001835:Body Weight / Q000187:drug effects / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D003720:Densitometry; D000229:Dependovirus / Q000235:genetics*; D004318:Doxycycline / Q000097:blood / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013752:Tetracycline / Q000494:pharmacology
chemical_list: D051100:Glial Cell Line-Derived Neurotrophic Factor; D013752:Tetracycline; D004318:Doxycycline
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