title: The promotion of functional recovery and nerve regeneration after spinal cord injury by lentiviral vectors encoding Lingo-1 shRNA delivered by Pluronic F-127.
abstract: Lingo-1 is selectively expressed on both oligodendrocytes and neurons in the central nervous system (CNS) and serves as a key negative regulator of nerve regeneration, implying a therapeutic target for spinal cord injury (SCI). Here we described a strategy to knock-down Lingo-1 expression in vivo using lentiviral vectors encoding Lingo-1 short harpin interfering RNA (shRNA) delivered by Pluronic F-127 (PF-127) gel, a non-cytotoxic scaffold and gene delivery carrier, after the complete transection of the T10 spinal cord in adult rats. We showed administration of PF-127 encapsulating Lingo-1 shRNA lentiviral vectors efficiently down-regulated the expression of Lingo-1, and exhibited transduction efﬁciency comparable to using vectors alone in oligodendrocyte culture in vitro. Furthermore, similar silencing effects and higher transfection efficiency were observed in vivo when Lingo-1 shRNA was co-delivered to the injured site by PF-127 gel with lower viral concentrations. Cografting of gel and Lingo-1 RNAi significantly promoted functional recovery and nerve regeneration, enhanced neurite outgrowth and synapses formation, preserved myelinated axons, and induced the proliferation of glial cells. In addition, the combined implantation also improved neuronal survival and inhibited cell apoptosis, which may be associated with the attenuation of endoplasmic reticulum (ER) stress after SCI. Together, our data indicated that delivering Lingo-1 shRNA by gel scaffold was a valuable treatment approach to SCI and PF-127 delivery of viral vectors to the spinal cord may provide strategy to study and develop therapies for SCI.
authors.affiliation: Research Center of Translational Medicine, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
authors.forename: Hong-Fu
authors.initials: HF
authors.lastname: Wu
authors.forename: Jing-Sheng
authors.initials: JS
authors.lastname: Cen
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhong
authors.forename: Luming
authors.initials: L
authors.lastname: Chen
authors.forename: Jue
authors.initials: J
authors.lastname: Wang
authors.forename: David Y B
authors.initials: DY
authors.lastname: Deng
authors.forename: Yong
authors.initials: Y
authors.lastname: Wan
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D005071:Evoked Potentials; D005260:Female; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D009416:Nerve Regeneration; D009419:Nerve Tissue Proteins / Q000235:genetics*; D020442:Poloxamer / Q000008:administration & dosage*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000008:administration & dosage*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013119:Spinal Cord Injuries / Q000503:physiopathology*
chemical_list: D017931:DNA Primers; C525164:LINGO1 protein, rat; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D020442:Poloxamer; D012313:RNA