Manumycin A corrects aberrant splicing of Clcn1 in myotonic dystrophy type 1 (DM1) mice

Oana, Kosuke; Oma, Yoko; Suo, Satoshi; Takahashi, Masanori; Nishino, Ichizo; Takeda, Shin’ichi; Ishiura, Shoichi

doi:10.1038/srep02142

Cited by 22 publications

(25 citation statements)

References 44 publications

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“…Manipulation of splicing by different means is considered a promising therapeutic approach to human disorders ( Havens et al, 2013 ; Singh and Cooper, 2012 ), such as for DM1 where target-based identification of active antisense oligonucleotides and rational design of small molecules are actively pursued strategies ( Wheeler et al, 2009 ; Childs-Disney et al, 2013 ). A different strategy is the development of high-throughput screening assays using cell culture systems to search for chemical modulators of specific missplicing events ( Havens et al, 2013 ), a technique hardly tested for DM1 ( O’Leary et al, 2010 ; Oana et al, 2013 ). Focusing on improving the often quite poor quality of hits resulting from traditional in vitro brute-force pharmacological screenings, the present work has established a novel in vivo screening methodology based on an automated platform for large-scale and cost-effective Drosophila -based evaluation of small molecules, pioneering the exploitation of a disease-linked alternative splicing reporter system.…”

Section: Discussionmentioning

confidence: 99%

Development of aDrosophila melanogasterspliceosensor system forin vivohigh-throughput screening in myotonic dystrophy type 1

Garcia-Alcover

Colonques-Bellmunt

Garijo

et al. 2014

Disease Models &Amp; Mechanisms

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Alternative splicing of pre-mRNAs is an important mechanism that regulates cellular function in higher eukaryotes. A growing number of human genetic diseases involve splicing defects that are directly connected to their pathology. In myotonic dystrophy type 1 (DM1), several clinical manifestations have been proposed to be the consequence of tissue-specific missplicing of numerous genes. These events are triggered by an RNA gain-of-function and resultant deregulation of specific RNA-binding factors, such as the nuclear sequestration of muscleblind-like family factors (MBNL1–MBNL3). Thus, the identification of chemical modulators of splicing events could lead to the development of the first valid therapy for DM1 patients. To this end, we have generated and validated transgenic flies that contain a luciferase-reporter-based system that is coupled to the expression of MBNL1-reliant splicing (spliceosensor flies), to assess events that are deregulated in DM1 patients in a relevant disease tissue. We then developed an innovative 96-well plate screening platform to carry out in vivo high-throughput pharmacological screening (HTS) with the spliceosensor model. After a large-scale evaluation (>16,000 chemical entities), several reliable splicing modulators (hits) were identified. Hit validation steps recognized separate DM1-linked therapeutic traits for some of the hits, which corroborated the feasibility of the approach described herein to reveal promising drug candidates to correct missplicing in DM1. This powerful Drosophila-based screening tool might also be applied in other disease models displaying abnormal alternative splicing, thus offering myriad uses in drug discovery.

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Section: Discussionmentioning

confidence: 99%

Development of aDrosophila melanogasterspliceosensor system forin vivohigh-throughput screening in myotonic dystrophy type 1

Garcia-Alcover

Colonques-Bellmunt

Garijo

et al. 2014

Disease Models &Amp; Mechanisms

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“…Generell sind Stabilität, Qualität und Lokalisation einer mRNA, der Export aus dem Kern und die Effizienz der Translation trotz der nicht-kodierenden Funktion dieses Abschnitts stark von der 3'UTR-Region abhängig [35]. Die Myotonie bei der DM1 ist durch gestörtes, aberrantes Splicing des muskulären Chlorid-Kanal-Gens (CLCN1) verursacht [36]. [44].…”

Section: Haut-und Hautanhangsgebildeunclassified

Myotone Dystrophien: Klinik, Pathogenese, Diagnostik und Therapie

Finsterer

Rudnik‐Schöneborn

2015

Fortschr Neurol Psychiatr

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The autosomal-dominant myotonic dystrophies dystrophia myotonica type-1 (DM1, Curschmann-Steinert disease) and dystrophia myotonica type-2 (DM2, proximal myotonic myopathy (PROMM)), are, contrary to the non-dystrophic myotonias, progressive multisystem disorders. DM1 and DM2 are the most frequent of the muscular dystrophies. In both diseases the skeletal muscle is the most severely affected organ (weakness, wasting, myotonia, myalgia). Additionally, they manifest in the eye, heart, brain, endocrine glands, gastrointestinal tract, skin, skeleton, and peripheral nerves. Phenotypes of DM1 may be classified as congenital, juvenile, classical, or late onset. DM2 is a disorder of the middle or older age and usually has a milder course compared to DM1. DM1 is due to a CTG-repeat expansion > 50 repeats in the non-coding 3' UTR of the DMPK-gene. DM2 is caused by a CCTG-repeat expansion to 75 - 11 000 repeats in intron-1 of the CNBP/ZNF9 gene. Mutant pre-mRNAs of both genes aggregate within the nucleus (nuclear foci), which sequester RNA-binding proteins and result in an abnormal protein expression via alternative splicing in downstream effector genes (toxic RNA diseases). Other mechanisms seem to play an additional pathogenetic role. Clinical severity of DM1 increases from generation to generation (anticipation). The higher the repeat expansion the more severe the DM1 phenotype. In DM2 severity of symptoms and age at onset do not correlate with the expansion size. Contrary to DM2, there is a congenital form and anticipation in DM1.

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“…Several other neurological and motor neuron disorders are linked by common dysfunction in RNA processing, including fused in sarcoma (FUS)amyotrophic lateral sclerosis (FUS-ALS), TAR DNA binding protein (TDP43)-ALS, frontotemporal dementia (FTD), spinal muscular atrophy (SMA) and autism spectrum disorders (Achsel et al 2013;Ling et al 2013;Corominas et al 2014;Xiong et al 2015). Approaches to normalize the RNA splicing events are therefore being explored as therapeutic strategies to compensate for diseases such as SMA (Hua et al 2011) and DM1 (Oana et al 2013;Wojciechowska et al 2014).…”

Section: Current Directions: Alternative Splicing Disease and Therapymentioning

confidence: 99%

Tracks through the genome to physiological events

Lipscombe

Pan

Schorge

2015

Experimental Physiology

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New Findings What is the topic of this review? We discuss tools available to access genome‐wide data sets that harbour cell‐specific, brain region‐specific and tissue‐specific information on exon usage for several species, including humans. In this Review, we demonstrate how to access this information in genome databases and its enormous value to physiology. What advances does it highlight? The sheer scale of protein diversity that is possible from complex genes, including those that encode voltage‐gated ion channels, is vast. But this choice is critical for a complete understanding of protein function in the most physiologically relevant context. Many proteins of great interest to physiologists and neuroscientists are structurally complex and located in specialized subcellular domains, such as neuronal synapses and transverse tubules of muscle. Genes that encode these critical signalling molecules (receptors, ion channels, transporters, enzymes, cell adhesion molecules, cell–cell interaction proteins and cytoskeletal proteins) are similarly complex. Typically, these genes are large; human Dystrophin (DMD) encodes a cytoskeletal protein of muscle and it is the largest naturally occurring gene at a staggering 2.3 Mb. Large genes contain many non‐coding introns and coding exons; human Titin (TTN), which encodes a protein essential for the assembly and functioning of vertebrate striated muscles, has over 350 exons and consequently has an enormous capacity to generate different forms of Titin mRNAs that have unique exon combinations. Functional and pharmacological differences among protein isoforms originating from the same gene may be subtle but nonetheless of critical physiological significance. Standard functional, immunological and pharmacological approaches, so useful for characterizing proteins encoded by different genes, typically fail to discriminate among splice isoforms of individual genes. Tools are now available to access genome‐wide data sets that harbour cell‐specific, brain region‐specific and tissue‐specific information on exon usage for several species, including humans. In this Review, we demonstrate how to access this information in genome databases and its enormous value to physiology.

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Manumycin A corrects aberrant splicing of Clcn1 in myotonic dystrophy type 1 (DM1) mice

Cited by 22 publications

References 44 publications

Development of aDrosophila melanogasterspliceosensor system forin vivohigh-throughput screening in myotonic dystrophy type 1

Development of aDrosophila melanogasterspliceosensor system forin vivohigh-throughput screening in myotonic dystrophy type 1

Myotone Dystrophien: Klinik, Pathogenese, Diagnostik und Therapie

Tracks through the genome to physiological events

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