Article thumbnail

Targeted Skipping of Human Dystrophin Exons in Transgenic Mouse Model Systemically for Antisense Drug Development

By Bo Wu, Ehsan Benrashid, Peijuan Lu, Caryn Cloer, Allen Zillmer, Mona Shaban and Qi Long Lu

Abstract

Antisense therapy has recently been demonstrated with great potential for targeted exon skipping and restoration of dystrophin production in cultured muscle cells and in muscles of Duchenne Muscular Dystrophy (DMD) patients. Therapeutic values of exon skipping critically depend on efficacy of the drugs, antisense oligomers (AOs). However, no animal model has been established to test AO targeting human dystrophin exon in vivo systemically. In this study, we applied Vivo-Morpholino to the hDMD/mdx mouse, a transgenic model carrying the full-length human dystrophin gene with mdx background, and achieved for the first time more than 70% efficiency of targeted human dystrophin exon skipping in vivo systemically. We also established a GFP-reporter myoblast culture to screen AOs targeting human dystrophin exon 50. Antisense efficiency for most AOs is consistent between the reporter cells, human myoblasts and in the hDMD/mdx mice in vivo. However, variation in efficiency was also clearly observed. A combination of in vitro cell culture and a Vivo-Morpholino based evaluation in vivo systemically in the hDMD/mdx mice therefore may represent a prudent approach for selecting AO drug and to meet the regulatory requirement

Topics: Research Article
Publisher: Public Library of Science
OAI identifier: oai:pubmedcentral.nih.gov:3096650
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (2009). AartsmaRus A
  2. (2010). Accurate quantification of dystrophin mRNA and exon skipping levels in duchenne muscular dystrophy.
  3. (2007). Antisense oligonucleotide-induced exon skipping across the human dystrophin gene transcript.
  4. (2010). Antisense-mediated modulation of splicing: therapeutic implications for Duchenne muscular dystrophy.
  5. (2001). Antisenseinduced exon skipping and synthesis of dystrophin in the mdx mouse.
  6. (1993). Characterization of translational frame exception patients in Duchenne/Becker muscular dystrophy.
  7. (1987). Dystrophin: the protein product of the Duchenne muscular dystrophy locus.
  8. (2008). Effective rescue of dystrophin improves cardiac function in dystrophin-deficient mice by a modified morpholino oligomer.
  9. (2009). Efficacy of systemic morpholino exon-skipping in Duchenne dystrophy dogs.
  10. (2010). Exonic sequences provide better targets for antisense oligonucleotides than splice site sequences in the modulation of Duchenne muscular dystrophy splicing.
  11. (2003). Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse.
  12. (2005). Functional analysis of 114 exon-internal AONs for targeted DMD exon skipping: indication for steric hindrance of SR protein binding sites.
  13. (2008). Generation and characterization of transgenic mice with the full-length human DMD gene.
  14. (1998). Giant dystrophin deletion associated with congenital cataract and mild muscular dystrophy.
  15. (2010). Guaninie Analogues Enhance Antisense Oligonucleotide-induced Exon Skipping in Dystrophin Gene In Vitro and In Vivo.
  16. (2009). Guidelines for antisense oligonucleotide design and insight into splicemodulating mechanisms.
  17. (2007). Local dystrophin restoration with antisense oligonucleotide PRO051.
  18. (2009). Local restoration of dystrophin expression with the morpholino oligomer AVI-4658 in Duchenne muscular dystrophy: a single-blind, placebo-controlled, dose-escalation, proof-of-concept study.
  19. (2010). mdx mouse model.
  20. (1998). Modification of splicing in the dystrophin gene in cultured Mdx muscle cells by antisense oligoribonucleotides.
  21. (2001). Nuclear antisense effects of neutral, anionic and cationic oligonucleotide analogs.
  22. (2001). Oligonucleotides against a splicing enhancer sequence led to dystrophin production in muscle cells from a Duchenne muscular dystrophy patient.
  23. (1993). Restoration of correct splicing in thalassemic premRNA by antisense of oligonucleotide.
  24. (2011). Systemic administration of PRO051 in Duchenne’s muscular dystrophy.
  25. (2005). Systemic delivery of antisense oligoribonucleotide restores dystrophin expression in bodywide skeletal muscles.
  26. (2006). Systemic delivery of morpholino oligonucleotide restores dystrophin expression bodywide and improves dystrophic pathology.
  27. (2004). Targeted exon skipping in transgenic hDMD mice: A model for direct preclinical screening of human-specific antisense oligonucleotides.
  28. (2011). Targeted Skipping of Human Dystrophin Exons PLoS ONE | www.plosone.org 10
  29. (2003). Therapeutic antisense-induced exon skipping in cultured muscle cells from six different DMD patients.
  30. (2006). Therapeutic modulation of DMD splicing by blocking exonic splicing enhancer sites with antisense oligonucleotides.
  31. (1990). Very mild muscular dystrophy associated with the deletion of 46% of dystrophin.