Reprogramming axonal behavior by axon-specific viral transduction

• Published in: Gene therapy Vol. 19; no. 9; pp. 947 - 955
• Main Authors: WALKER, B. A, HENGST, U, KIM, H. J, JEON, N. L, SCHMIDT, E. F, HEINTZ, N, MILNER, T. A, JAFFREY, S. R
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group 01.09.2012
• Subjects: Adenylyl Cyclases - genetics; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animal experimentation; Animals; Applied cell therapy and gene therapy; Axon guidance; Axons; Axons - metabolism; Axons - physiology; Biological and medical sciences; Biotechnology; Care and treatment; Cell body; Epitopes; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Targeting - methods; Gene therapy; Genetic engineering; Genetic Vectors; Genomes; Health aspects; Health. Pharmaceutical industry; Industrial applications and implications. Economical aspects; Injuries; Medical sciences; Mice; Nerve Regeneration; Neurodegeneration; Neurological disorders; Peripheral nerve diseases; Physiological aspects; Protein expression; Proteins; Proteomes; Ribosomes; Ribosomes - virology; RNA viruses; Rodents; Signal transduction; Sindbis virus; Sindbis Virus - genetics; Spinal Cord; Transcription; Transduction; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Translation; Cellular reprogramming; Regeneration; Neuron; Recombinant virus; viral vector gene transfer; Axon; axon regeneration; Transduction; Behavior; Gene therapy; Genetic transfer; Sindbis
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2011.217

The treatment of axonal disorders, such as diseases associated with axonal injury and degeneration, is limited by the inability to directly target therapeutic protein expression to injured axons. Current gene therapy approaches rely on infection and transcription of viral genes in the cell body. Here, we describe an approach to target gene expression selectively to axons. Using a genetically engineered mouse containing epitope-labeled ribosomes, we find that neurons in adult animals contain ribosomes in distal axons. To use axonal ribosomes to alter local protein expression, we utilized a Sindbis virus containing an RNA genome that has been modified so that it can be directly used as a template for translation. Selective application of this virus to axons leads to local translation of heterologous proteins. Furthermore, we demonstrate that selective axonal protein expression can be used to modify axonal signaling in cultured neurons, enabling axons to grow over inhibitory substrates typically encountered following axonal injury. We also show that this viral approach also can be used to achieve heterologous expression in axons of living animals, indicating that this approach can be used to alter the axonal proteome in vivo. Together, these data identify a novel strategy to manipulate protein expression in axons, and provides a novel approach for using gene therapies for disorders of axonal function.