Actinomycin D Specifically Reduces Expanded CUG Repeat RNA in Myotonic Dystrophy Models

Ruth B Siboni; Masayuki Nakamori; Stacey D Wagner; Adam J Struck; Leslie A Coonrod; Shanee A Harriott; Daniel M Cass; Matthew K Tanner; J Andrew Berglund

doi:10.1016/j.celrep.2015.11.028

Actinomycin D Specifically Reduces Expanded CUG Repeat RNA in Myotonic Dystrophy Models

Cell Rep. 2015 Dec 22;13(11):2386-2394. doi: 10.1016/j.celrep.2015.11.028. Epub 2015 Dec 10.

Authors

Ruth B Siboni¹, Masayuki Nakamori², Stacey D Wagner¹, Adam J Struck¹, Leslie A Coonrod¹, Shanee A Harriott³, Daniel M Cass³, Matthew K Tanner¹, J Andrew Berglund⁴

Affiliations

¹ Institute of Molecular Biology and Department of Chemistry and Biochemistry, University of Oregon, Eugene, OR 97403, USA.
² Department of Neurology, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
³ Department of Chemistry, Reed College, Portland, OR 97202, USA.
⁴ Institute of Molecular Biology and Department of Chemistry and Biochemistry, University of Oregon, Eugene, OR 97403, USA; Department of Biochemistry & Molecular Biology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA. Electronic address: [email protected].

Abstract

Myotonic dystrophy type 1 (DM1) is an inherited disease characterized by the inability to relax contracted muscles. Affected individuals carry large CTG expansions that are toxic when transcribed. One possible treatment approach is to reduce or eliminate transcription of CTG repeats. Actinomycin D (ActD) is a potent transcription inhibitor and FDA-approved chemotherapeutic that binds GC-rich DNA with high affinity. Here, we report that ActD decreased CUG transcript levels in a dose-dependent manner in DM1 cell and mouse models at significantly lower concentrations (nanomolar) compared to its use as a general transcription inhibitor or chemotherapeutic. ActD also significantly reversed DM1-associated splicing defects in a DM1 mouse model, and did so within the currently approved human treatment range. RNA-seq analyses showed that low concentrations of ActD did not globally inhibit transcription in a DM1 mouse model. These results indicate that transcription inhibition of CTG expansions is a promising treatment approach for DM1.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Autophagy-Related Proteins
Base Sequence
Calorimetry
Chloride Channels / genetics
Chloride Channels / metabolism
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Dactinomycin / pharmacology*
Disease Models, Animal
Guanine Nucleotide Exchange Factors / genetics
Guanine Nucleotide Exchange Factors / metabolism
HeLa Cells
Humans
Mice
Microscopy, Fluorescence
Myotonic Dystrophy / metabolism
Myotonic Dystrophy / pathology*
RNA / chemistry
RNA / metabolism*
RNA Splicing / drug effects
RNA, Messenger / metabolism
RNA-Binding Proteins / genetics
RNA-Binding Proteins / metabolism
Sarcoplasmic Reticulum Calcium-Transporting ATPases / genetics
Sarcoplasmic Reticulum Calcium-Transporting ATPases / metabolism
Sequence Analysis, RNA
Transcription, Genetic / drug effects
Trinucleotide Repeat Expansion / drug effects*
Trinucleotide Repeat Expansion / genetics
Vesicular Transport Proteins

Substances

Atp2a1 protein, mouse
Autophagy-Related Proteins
CLC-1 channel
Chloride Channels
DNA-Binding Proteins
Guanine Nucleotide Exchange Factors
Mbnl1 protein, mouse
RNA, Messenger
RNA-Binding Proteins
VPS39 protein, mouse
Vesicular Transport Proteins
Dactinomycin
RNA
Sarcoplasmic Reticulum Calcium-Transporting ATPases

Abstract

Publication types

MeSH terms

Substances

Grants and funding