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DMD pseudoexon mutations: splicing efficiency, phenotype, and potential therapy.
Ann Neurol. 2008 Jan; 63(1):81-9.AN

Abstract

OBJECTIVE

The degenerative muscle diseases Duchenne (DMD) and Becker muscular dystrophy result from mutations in the DMD gene, which encodes the dystrophin protein. Recent improvements in mutational analysis techniques have resulted in the increasing identification of deep intronic point mutations, which alter splicing such that intronic sequences are included in the messenger RNA as "pseudoexons." We sought to test the hypothesis that the clinical phenotype correlates with splicing efficiency of these mutations, and to test the feasibility of antisense oligonucleotide (AON)-mediated pseudoexon skipping.

METHODS

We identified three pseudoexon insertion mutations in dystrophinopathy patients, two of whom had tissue available for further analysis. For these two out-of-frame pseudoexon mutations (one associated with Becker muscular dystrophy and one with DMD), mutation-induced splicing was tested by quantitative reverse transcription polymerase chain reaction; pseudoexon skipping was tested using AONs composed of 2'-O-methyl-modified bases on a phosphorothioate backbone to treat cultured primary myoblasts.

RESULTS

Variable amounts of pseudoexon inclusion correlates with the severity of the dystrophinopathy phenotype in these two patients. AON treatment directed at the pseudoexon results in the expression of full-length dystrophin in a DMD myoblast line.

INTERPRETATION

Both DMD and Becker muscular dystrophy can result from out-of-frame pseudoexons, with the difference in phenotype being due to variable efficiency of the newly generated splicing signal. AON-mediated pseudoexon skipping therapy is a viable approach to these patients and would be predicted to result in increased expression of wild-type dystrophin protein.

Authors+Show Affiliations

Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Case Reports
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18059005

Citation

Gurvich, Olga L., et al. "DMD Pseudoexon Mutations: Splicing Efficiency, Phenotype, and Potential Therapy." Annals of Neurology, vol. 63, no. 1, 2008, pp. 81-9.
Gurvich OL, Tuohy TM, Howard MT, et al. DMD pseudoexon mutations: splicing efficiency, phenotype, and potential therapy. Ann Neurol. 2008;63(1):81-9.
Gurvich, O. L., Tuohy, T. M., Howard, M. T., Finkel, R. S., Medne, L., Anderson, C. B., Weiss, R. B., Wilton, S. D., & Flanigan, K. M. (2008). DMD pseudoexon mutations: splicing efficiency, phenotype, and potential therapy. Annals of Neurology, 63(1), 81-9.
Gurvich OL, et al. DMD Pseudoexon Mutations: Splicing Efficiency, Phenotype, and Potential Therapy. Ann Neurol. 2008;63(1):81-9. PubMed PMID: 18059005.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - DMD pseudoexon mutations: splicing efficiency, phenotype, and potential therapy. AU - Gurvich,Olga L, AU - Tuohy,Therese M, AU - Howard,Michael T, AU - Finkel,Richard S, AU - Medne,Livija, AU - Anderson,Christine B, AU - Weiss,Robert B, AU - Wilton,Steve D, AU - Flanigan,Kevin M, PY - 2007/12/7/pubmed PY - 2008/3/13/medline PY - 2007/12/7/entrez SP - 81 EP - 9 JF - Annals of neurology JO - Ann Neurol VL - 63 IS - 1 N2 - OBJECTIVE: The degenerative muscle diseases Duchenne (DMD) and Becker muscular dystrophy result from mutations in the DMD gene, which encodes the dystrophin protein. Recent improvements in mutational analysis techniques have resulted in the increasing identification of deep intronic point mutations, which alter splicing such that intronic sequences are included in the messenger RNA as "pseudoexons." We sought to test the hypothesis that the clinical phenotype correlates with splicing efficiency of these mutations, and to test the feasibility of antisense oligonucleotide (AON)-mediated pseudoexon skipping. METHODS: We identified three pseudoexon insertion mutations in dystrophinopathy patients, two of whom had tissue available for further analysis. For these two out-of-frame pseudoexon mutations (one associated with Becker muscular dystrophy and one with DMD), mutation-induced splicing was tested by quantitative reverse transcription polymerase chain reaction; pseudoexon skipping was tested using AONs composed of 2'-O-methyl-modified bases on a phosphorothioate backbone to treat cultured primary myoblasts. RESULTS: Variable amounts of pseudoexon inclusion correlates with the severity of the dystrophinopathy phenotype in these two patients. AON treatment directed at the pseudoexon results in the expression of full-length dystrophin in a DMD myoblast line. INTERPRETATION: Both DMD and Becker muscular dystrophy can result from out-of-frame pseudoexons, with the difference in phenotype being due to variable efficiency of the newly generated splicing signal. AON-mediated pseudoexon skipping therapy is a viable approach to these patients and would be predicted to result in increased expression of wild-type dystrophin protein. SN - 1531-8249 UR - https://www.unboundmedicine.com/medline/citation/18059005/DMD_pseudoexon_mutations:_splicing_efficiency_phenotype_and_potential_therapy_ L2 - https://doi.org/10.1002/ana.21290 DB - PRIME DP - Unbound Medicine ER -