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Treatment of DMD 5’ Mutations through Two Different Exon 2 Skipping Strategies: rAAV9.U7snRNA Mediated Skipping and Antisense Morpholino Oligomers

Simmons, Tabatha Renee
http://orcid.org/0000-0002-4457-3623
http://rave.ohiolink.edu/etdc/view?acc_num=osu1469122227

Abstract Details

2016, Doctor of Philosophy, Ohio State University, Molecular, Cellular and Developmental Biology.
Duchenne muscular dystrophy (DMD) is an X-linked progressive disease characterized by loss of dystrophin protein as a result of truncating mutations in the DMD gene. Initial exon-skipping therapies have sought to treat deletion mutations that abolish an open reading frame (ORF) because when skipped, an ORF can be restored and produce an internally deleted, yet partially functional protein. In contrast, skipping of one copy of a duplicated exon would be expected to result in a full-length transcript and production of a wild type protein. To test this approach, we have developed a mouse model which contains a duplication of exon 2 (Dup2), the most common single exon duplication in patients. We are developing exon-skipping therapies for duplication mutations, which account for around 6%-11% of all duplication mutations by testing both virally (AAV) mediated skipping induced by a modified U7snRNA (rAAV9.U7.ACCA) and antisense oligomer-induced skipping in the Dup2 mouse. Dose escalation studies of both intramuscular and systemic delivery of scAAV9.U7.ACCA show efficient skipping of exon 2, along with increased expression of properly localized dystrophin that restores muscle function. Furthermore, intramuscular injections of antisense oligomers results in widespread skipping and properly localized dystrophin protein. These data suggest that skipping of a duplicated exon 2 may be a feasible therapeutic approach, particularly because skipping of exon 2 may be associated with an apparently unlimited therapeutic window. Complete skipping of exon 2 results in activation of an internal ribosome entry site (IRES) located in exon 5 of dystrophin allowing for cap-independent translation from an alternative initiation site within exon 6. Each of the approximately 5% of patients harboring mutations 5’ of the IRES in exon 5 have the potential to benefit from this highly functional dystrophin isoform.
Kevin Flanigan (Advisor)
Scott Harper (Committee Member)
Louise Rodino-Klapac (Committee Member)
Denis Guttridge (Committee Member)
125 p.
Biomedical Research
Duchenne Muscular Dystrophy, Exon Skipping, U7snRNA, Antisense Oligomer

Recommended Citations

Citations

  • Simmons, T. R. (2016). Treatment of DMD 5’ Mutations through Two Different Exon 2 Skipping Strategies: rAAV9.U7snRNA Mediated Skipping and Antisense Morpholino Oligomers [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1469122227

    APA Style (7th edition)

  • Simmons, Tabatha. Treatment of DMD 5’ Mutations through Two Different Exon 2 Skipping Strategies: rAAV9.U7snRNA Mediated Skipping and Antisense Morpholino Oligomers. 2016. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1469122227.

    MLA Style (8th edition)

  • Simmons, Tabatha. "Treatment of DMD 5’ Mutations through Two Different Exon 2 Skipping Strategies: rAAV9.U7snRNA Mediated Skipping and Antisense Morpholino Oligomers." Doctoral dissertation, Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1469122227

    Chicago Manual of Style (17th edition)

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This open access ETD is published by The Ohio State University and OhioLINK.