Tight Long-term dynamic doxycycline responsive nigrostriatal GDNF using a single rAAV vector.
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.
Manfredsson
Fredric P
FP
Department of Neuroscience, Powell Gene Therapy Center, McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida 32611, USA.
Burger
Corinna
C
Rising
Aaron C
AC
Zuobi-Hasona
Kheir
K
Sullivan
Layla F
LF
Lewin
Alfred S
AS
Huang
Julia
J
Piercefield
Emily
E
Muzyczka
Nicholas
N
Mandel
Ronald J
RJ
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
D051100:Glial Cell Line-Derived Neurotrophic Factor; D013752:Tetracycline; D004318:Doxycycline
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