title: Transgene expression in the striatum following intracerebral injections of DNA nanoparticles encoding for human glial cell line-derived neurotrophic factor.
abstract: A goal of our studies is to develop a potential therapeutic for Parkinson's disease (PD) by a human glial cell line-derived neurotrophic factor (hGDNF) expression plasmid administered to the rat striatum as a compacted DNA nanoparticle (DNP) and which will generate long-term hGDNF expression at biologically active levels. In the present study, we used a DNA plasmid encoding for hGDNF and a polyubiquitin C (UbC) promoter that was previously shown to have activity in both neurons and glia, but primarily in glia. A two-fold improvement was observed at the highest plasmid dose when using hGDNF DNA incorporating sequences found in RNA splice variant 1 compared with splice variant 2; of note, the splice variant 2 sequence is used in most preclinical studies. This optimized expression cassette design includes flanking scaffold matrix attachment elements (S/MARs) as well as a CpG-depleted prokaryotic domain and, where possible, eukaryotic elements. Stable long-term GDNF activity at levels 300-400% higher than baseline was observed following a single intracerebral injection. In a previous study, DNP plasmids encoding for reporter genes had been successful in generating long-term reporter transgene activity in the striatum (>365 days) and in this study produced sustained GDNF activity at the longest assessed time point (6 months).
authors.affiliation: Department of Neurosurgery, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Fletcher
authors.forename: T H
authors.initials: TH
authors.lastname: Kowalczyk
authors.forename: L
authors.initials: L
authors.lastname: Padegimas
authors.forename: M J
authors.initials: MJ
authors.lastname: Cooper
authors.forename: D M
authors.initials: DM
authors.lastname: Yurek
mesh_terms: D000818:Animals; D001923:Brain Chemistry / Q000235:genetics*; D003342:Corpus Striatum / Q000378:metabolism*; D004247:DNA / Q000008:administration & dosage*; D004195:Disease Models, Animal; D016503:Drug Delivery Systems / Q000379:methods; D005786:Gene Expression Regulation / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000008:administration & dosage / Q000235:genetics*; D006801:Humans; D008297:Male; D008845:Microinjections / Q000379:methods; D053758:Nanoparticles / Q000008:administration & dosage* / Q000627:therapeutic use; D020734:Parkinsonian Disorders / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D051100:Glial Cell Line-Derived Neurotrophic Factor; D004247:DNA