title: Regulation of human GDNF gene expression in nigral dopaminergic neurons using a new doxycycline-regulated NTS-polyplex nanoparticle system.
abstract: The human glial-cell derived neurotrophic factor (hGDNF) gene transfer by neurotensin (NTS)-polyplex nanoparticles functionally restores the dopamine nigrostriatal system in experimental Parkinson's disease models. However, high levels of sustained expression of GDNF eventually can cause harmful effects. Herein, we report an improved NTS-polyplex nanoparticle system that enables regulation of hGDNF expression within dopaminergic neurons. We constructed NTS-polyplex nanoparticles containing a single bifunctional plasmid that codes for the reverse tetracycline-controlled transactivator advanced (rtTA-Adv) under the control of NBRE3x promoter, and for hGDNF under the control of tetracycline-response element (TRE). Another bifunctional plasmid contained the enhanced green fluorescent protein (GFP) gene. Transient transfection experiments in N1E-115-Nurr1 cells showed that doxycycline (100 ng/mL) activates hGDNF and GFP expression. Doxycycline (5 mg/kg, i.p.) administration in rats activated hGDNF expression only in transfected dopaminergic neurons, whereas doxycycline withdrawal silenced transgene expression. Our results offer a specific doxycycline-regulated system suitable for nanomedicine-based treatment of Parkinson's disease.
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico.
authors.forename: Armando J
authors.initials: AJ
authors.lastname: Espadas-Alvarez
authors.affiliation: Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Bannon
authors.affiliation: CONACYT - Medical Research Unit in Neurological Diseases, National Medical Center "Siglo XXI", IMSS, Mexico City, Mexico.
authors.forename: Carlos E
authors.initials: CE
authors.lastname: Orozco-Barrios
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico.
authors.forename: Lourdes
authors.initials: L
authors.lastname: Escobedo-Sanchez
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico.
authors.forename: Jose
authors.initials: J
authors.lastname: Ayala-Davila
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico.
authors.forename: David
authors.initials: D
authors.lastname: Reyes-Corona
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico.
authors.forename: Guadalupe
authors.initials: G
authors.lastname: Soto-Rodriguez
authors.affiliation: Department of Toxicology, CINVESTAV, Mexico City, Mexico.
authors.forename: Vicente
authors.initials: V
authors.lastname: Escamilla-Rivera
authors.affiliation: Department of Toxicology, CINVESTAV, Mexico City, Mexico.
authors.forename: Andrea
authors.initials: A
authors.lastname: De Vizcaya-Ruiz
authors.affiliation: Department of Biosciences and Engineering, CIIEMAD-IPN, Mexico City, Mexico.
authors.forename: M
authors.initials: M
authors.lastname: Eugenia Gutierrez-Castillo
authors.affiliation: Knowledge transfer and commercialization office, the 3C agency, CINVESTAV, Mexico City, Mexico.
authors.forename: America
authors.initials: A
authors.lastname: Padilla-Viveros
authors.affiliation: Department of Physiology, Biophysics and Neurosciences, CINVESTAV, Mexico City, Mexico; PhD Program on Nanoscience and Nanotechnology (DNyN), CINVESTAV, Mexico City, Mexico. Electronic address: martinez.fong@gmail.com.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martinez-Fong
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D059290:Dopaminergic Neurons / Q000378:metabolism*; D004318:Doxycycline / Q000494:pharmacology*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D009496:Neurotensin / Q000737:chemistry*; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1 / Q000235:genetics* / Q000378:metabolism; D010300:Parkinson Disease / Q000188:drug therapy; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D020218:Response Elements; D014162:Transfection; D019076:Transgenes
chemical_list: C539950:NR6A1 protein, human; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1; D009496:Neurotensin; D004318:Doxycycline
keywords: Bifunctional plasmid; NBRE3x promoter; Tet-on system, gene delivery, nonviral vectors