title: Tf-lipoplex-mediated c-Jun silencing improves neuronal survival following excitotoxic damage in vivo.
abstract: Excitotoxicity is one of the main features responsible for neuronal cell death after acute brain injury and in several neurodegenerative disorders, for which only few therapeutic options are currently available. In this work, RNA interference was employed to identify and validate a potential target for successful treatment of excitotoxic brain injury, the transcription factor c-Jun. The nuclear translocation of c-Jun and its upregulation are early events following glutamate-induced excitotoxic damage in primary neuronal cultures. We present evidence for the efficient knockdown of this transcription factor using a non-viral vector consisting of cationic liposomes associated to transferrin (Tf-lipoplexes). Tf-lipoplexes were able to deliver anti-c-Jun siRNAs to neuronal cells in culture, resulting in efficient silencing of c-Jun mRNA and protein and in a significant decrease of cell death following glutamate-induced damage or oxygen-glucose deprivation. This formulation also leads to a significant c-Jun knockdown in the mouse hippocampus in vivo, resulting in the attenuation of both neuronal death and inflammation following kainic acid-mediated lesion of this region. Furthermore, a strong reduction of seizure activity and cytokine production was observed in animals treated with anti-c-Jun siRNAs. These findings demonstrate the efficient delivery of therapeutic siRNAs to the brain by Tf-lipoplexes and validate c-Jun as a promising therapeutic target in neurodegenerative disorders involving excitotoxic lesions.
authors.affiliation: Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal.
authors.forename: A L C
authors.initials: AL
authors.lastname: Cardoso
authors.forename: P
authors.initials: P
authors.lastname: Costa
authors.forename: L P
authors.initials: LP
authors.lastname: de Almeida
authors.forename: S
authors.initials: S
authors.lastname: Simões
authors.forename: N
authors.initials: N
authors.lastname: Plesnila
authors.forename: C
authors.initials: C
authors.lastname: Culmsee
authors.forename: E
authors.initials: E
authors.lastname: Wagner
authors.forename: M C Pedroso
authors.initials: MC
authors.lastname: de Lima
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002784:Cholesterol / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry*; D004339:Drug Compounding; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D020868:Gene Silencing / Q000187:drug effects*; D018698:Glutamic Acid / Q000633:toxicity; D006801:Humans; D007150:Immunohistochemistry; D007608:Kainic Acid / Q000633:toxicity; D008081:Liposomes; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D018696:Neuroprotective Agents / Q000008:administration & dosage* / Q000494:pharmacology / Q000627:therapeutic use; D021381:Protein Transport; D016755:Proto-Oncogene Proteins c-jun / Q000037:antagonists & inhibitors / Q000235:genetics*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000494:pharmacology / Q000627:therapeutic use; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012640:Seizures / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D014168:Transferrin / Q000737:chemistry*
chemical_list: D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D018696:Neuroprotective Agents; D016755:Proto-Oncogene Proteins c-jun; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; D014168:Transferrin; D018698:Glutamic Acid; D002784:Cholesterol; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; D007608:Kainic Acid