title: Retargeting of adenoviral vectors to neurons using the Hc fragment of tetanus toxin.
abstract: The Hc fragment of tetanus toxin (Hc) retains the specific nerve cell binding and transport properties of the holotoxin, but lacks any toxicity. We are investigating the potential for utilising its neurotropism for targeted gene delivery to the central nervous system. Previously we reported the use of Hc-polylysine conjugates for selective gene transfer into neuronal cells in vitro. However, as attempts to apply these constructs in vivo were not successful, we have extended these studies to modification of the tropism of adenoviral vectors. Either Hc-polylysine conjugates or the Fab fragment of a neutralising anti-knob antibody covalently bound to Hc were attached to the virus. Infection of neuronal and non-neuronal cell lines with retargeted virus showed highly increased neuronal cell selectivity, but no significant enhancement of gene delivery into these cells. High concentrations of free Hc blocked the infectivity of the retargeted vector efficiently. Intramuscular injection of retargeted virus into mouse tongues resulted in selective gene transfer to the neurons of the hypoglossal nucleus, where no pathological changes were observed. As differentiated neurons do not undergo cell division, appropriate vectors carrying a thymidine kinase gene, which allows selective elimination of dividing cells, may be exploitable for the treatment of tumours of the central nervous system. The demonstrated suitability of the Hc fragment of tetanus toxin as targeting moiety for viral vectors also indicates a potential for gene therapy of inherited neurodegenerative diseases such as spinal muscular atrophy.
authors.affiliation: Division of Biomedical Sciences, Imperial College School of Medicine, London, UK.
authors.forename: H
authors.initials: H
authors.lastname: Schneider
authors.forename: M
authors.initials: M
authors.lastname: Groves
authors.forename: C
authors.initials: C
authors.lastname: Mühle
authors.forename: P N
authors.initials: PN
authors.lastname: Reynolds
authors.forename: A
authors.initials: A
authors.lastname: Knight
authors.forename: M
authors.initials: M
authors.lastname: Themis
authors.forename: J
authors.initials: J
authors.lastname: Carvajal
authors.forename: F
authors.initials: F
authors.lastname: Scaravilli
authors.forename: D T
authors.initials: DT
authors.lastname: Curiel
authors.forename: N F
authors.initials: NF
authors.lastname: Fairweather
authors.forename: C
authors.initials: C
authors.lastname: Coutelle
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002493:Central Nervous System Diseases / Q000628:therapy*; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006651:Histocytochemistry; D006825:Hybridomas; D007267:Injections; D051379:Mice; D008815:Mice, Inbred Strains; D009046:Motor Neurons; D010446:Peptide Fragments; D011107:Polylysine; D051381:Rats; D013744:Tetanus Toxin; D014059:Tongue; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
chemical_list: D010446:Peptide Fragments; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C; D011107:Polylysine; D001616:beta-Galactosidase