title: Intramuscular injection of AAV-GDNF results in sustained expression of transgenic GDNF, and its delivery to spinal motoneurons by retrograde transport.
abstract: Adeno-associated virus (AAV) vector has been developed as an attractive gene delivery system with proven safety. Glial cell line-derived neurotrophic factor (GDNF) is proposed to be a promising therapeutic agent for amyotrophic lateral sclerosis (ALS) and other motor neuron diseases. The purpose of this report was to investigate transgenic GDNF expression at different time points post AAV mediated GDNF intramuscular delivery. An AAV vector was constructed to encode a recombinant fusion of GDNF tagged with a FLAG sequence at the C-terminal (AAV-GDNF) to distinguish it from its endogenous counterpart. A single intramuscular injection of AAV-GDNF led to substantial expression of transgenic GDNF which remained for at least 10 months in transduced gastrocnemius muscle. This transgenic GDNF was distributed in a large number of myofibers, mainly in the vicinity of the sarcolemma and predominantly concentrated at the sites of neuromuscular junctions (NMJs). Furthermore, transgenic GDNF, but not beta-galactosidase expressed as a control, was detected in the motoneurons that projected axons to the injected muscles, thus, indicating retrograde axonal transportation of the transgenic GDNF. This study provides a basis for a strategy of intramuscular AAV-GDNF delivery to protect motoneurons as a possible means of ALS treatment.
authors.affiliation: Division of Neurology, Department of Medicine, Jichi Medical School, 3311-1 Yakushiji, Minamikawachi-machi, Tochigi 329-0498, Japan.
authors.forename: Yan-Yan
authors.initials: YY
authors.lastname: Lu
authors.forename: Li-Jun
authors.initials: LJ
authors.lastname: Wang
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Muramatsu
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Ikeguchi
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Fujimoto
authors.forename: Takashi
authors.initials: T
authors.lastname: Okada
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mizukami
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsushita
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hanazono
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kume
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Nagatsu
authors.forename: Keiya
authors.initials: K
authors.lastname: Ozawa
authors.forename: Imaharu
authors.initials: I
authors.lastname: Nakano
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006801:Humans; D007150:Immunohistochemistry; D007273:Injections, Intramuscular; D051379:Mice; D009046:Motor Neurons / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism; D009414:Nerve Growth Factors / Q000096:biosynthesis* / Q000235:genetics*; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes
chemical_list: C495039:GDNF protein, human; C495040:Gdnf protein, mouse; D051100:Glial Cell Line-Derived Neurotrophic Factor; D009414:Nerve Growth Factors