Strategy for treating motor neuron diseases using a fusion protein of botulinum toxin binding domain and streptavidin for viral vector access: Work in progress

B. Daniel Drachman, Robert N. Adams, Uma Balasubramanian, Lu Yang

Research output: Contribution to journalArticle

Abstract

Although advances in understanding of the pathogenesis of amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) have suggested attractive treatment strategies, delivery of agents to motor neurons embedded within the spinal cord is problematic. We have designed a strategy based on the specificity of botulinum toxin, to direct entry of viral vectors carrying candidate therapeutic genes into motor neurons. We have engineered and expressed fusion proteins consisting of the binding domain of botulinum toxin type A fused to streptavidin (SAv). This fusion protein will direct viral vectors carrying therapeutic genes into motor nerve terminals where they biotinylated can enter the acidified endosomal compartments, be released and undergo retrograde transport, to deliver the genes to motor neurons. Both ends of the fusion proteins are shown to be functionally intact. The binding domain end binds to mammalian nerve terminals at neuromuscular junctions, ganglioside GT1b (a target of botulinum toxin), and a variety of neuronal cells including primary chick embryo motor neurons, N2A neuroblastoma cells, NG108-15 cells, but not to NG CR72 cells, which lack complex gangliosides. The streptavidin end binds to biotin, and to a biotinylated Alexa 488 fluorescent tag. Further studies are in progress to evaluate the delivery of genes to motor neurons in vivo, by the use of biotinylated viral vectors.

Original languageEnglish (US)
Pages (from-to)2872-2889
Number of pages18
JournalToxins
Volume2
Issue number12
DOIs
StatePublished - Dec 2010

Keywords

  • Als
  • Binding domain
  • Botulinum toxin
  • Gene transfer
  • Motor neuron diseases
  • Motor neurons
  • Sma
  • Therapeutic targeting
  • Viral vectors

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

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