FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2616392786> ?p ?o ?g. }

• W2616392786 endingPage "474" @default.
• W2616392786 startingPage "465" @default.
• W2616392786 abstract "Myotonic dystrophy type 1 (DM1), a dominant hereditary muscular dystrophy, is caused by an abnormal expansion of a (CTG)n trinucleotide repeat in the 3′ UTR of the human dystrophia myotonica protein kinase (DMPK) gene. As a consequence, mutant transcripts containing expanded CUG repeats are retained in nuclear foci and alter the function of splicing regulatory factors members of the MBNL and CELF families, resulting in alternative splicing misregulation of specific transcripts in affected DM1 tissues. In the present study, we treated DMSXL mice systemically with a 2′-4′-constrained, ethyl-modified (ISIS 486178) antisense oligonucleotide (ASO) targeted to the 3′ UTR of the DMPK gene, which led to a 70% reduction in CUGexp RNA abundance and foci in different skeletal muscles and a 30% reduction in the heart. Furthermore, treatment with ISIS 486178 ASO improved body weight, muscle strength, and muscle histology, whereas no overt toxicity was detected. This is evidence that the reduction of CUGexp RNA improves muscle strength in DM1, suggesting that muscle weakness in DM1 patients may be improved following elimination of toxic RNAs. Myotonic dystrophy type 1 (DM1), a dominant hereditary muscular dystrophy, is caused by an abnormal expansion of a (CTG)n trinucleotide repeat in the 3′ UTR of the human dystrophia myotonica protein kinase (DMPK) gene. As a consequence, mutant transcripts containing expanded CUG repeats are retained in nuclear foci and alter the function of splicing regulatory factors members of the MBNL and CELF families, resulting in alternative splicing misregulation of specific transcripts in affected DM1 tissues. In the present study, we treated DMSXL mice systemically with a 2′-4′-constrained, ethyl-modified (ISIS 486178) antisense oligonucleotide (ASO) targeted to the 3′ UTR of the DMPK gene, which led to a 70% reduction in CUGexp RNA abundance and foci in different skeletal muscles and a 30% reduction in the heart. Furthermore, treatment with ISIS 486178 ASO improved body weight, muscle strength, and muscle histology, whereas no overt toxicity was detected. This is evidence that the reduction of CUGexp RNA improves muscle strength in DM1, suggesting that muscle weakness in DM1 patients may be improved following elimination of toxic RNAs. Myotonic dystrophy type 1 (DM1), the most common form of muscular dystrophy in adults, with an incidence of 1:15,000, is a multisystemic disorder characterized by myotonia, progressive muscle wasting, cardiac conduction defects, as well as endocrine deficiencies and cognitive impairments.1Harper P.S. Myotonic Dystrophy. Saunders, 2001Google Scholar A severe congenital form (CDM1) of this disease is characterized by neonatal hypotonia, facial weakness, respiratory and feeding difficulties, and high neonatal mortality (16% versus 19:1,000 in the population). Available data suggest that CDM1 seems to be caused by delayed skeletal muscle and brain maturation,1Harper P.S. Myotonic Dystrophy. Saunders, 2001Google Scholar, 2Farkas-Bargeton E. Barbet J.P. Dancea S. Wehrle R. Checouri A. Dulac O. Immaturity of muscle fibers in the congenital form of myotonic dystrophy: its consequences and its origin.J. Neurol. Sci. 1988; 83: 145-159Abstract Full Text PDF PubMed Scopus (66) Google Scholar, 3Furling D. Coiffier L. Mouly V. Barbet J.P. St Guily J.L. Taneja K. Gourdon G. Junien C. Butler-Browne G.S. Defective satellite cells in congenital myotonic dystrophy.Hum. Mol. Genet. 2001; 10: 2079-2087Crossref PubMed Scopus (97) Google Scholar, 4Sarnat H.B. Silbert S.W. Maturational arrest of fetal muscle in neonatal myotonic dystrophy. A pathologic study of four cases.Arch. Neurol. 1976; 33: 466-474Crossref PubMed Scopus (110) Google Scholar whereas the adult form of DM1 appears to result from a degenerative process. DM1 is caused by an expanded (CTG)n repeat, where n varies from 50 to several thousand, in the 3′ UTR of the dystrophia myotonica protein kinase (DMPK) gene.5Thornton C.A. Myotonic dystrophy.Neurol. Clin. 2014; 32: 705-719Abstract Full Text Full Text PDF PubMed Scopus (230) Google Scholar The most commonly accepted mechanistic explanation is that the nuclear accumulation of transcripts containing CUG expansions sequesters the RNA-binding protein MBNL1 and stabilizes CELF1 via hyperphosphorylation.6Taneja K.L. McCurrach M. Schalling M. Housman D. Singer R.H. Foci of trinucleotide repeat transcripts in nuclei of myotonic dystrophy cells and tissues.J. Cell Biol. 1995; 128: 995-1002Crossref PubMed Scopus (500) Google Scholar, 7Kalsotra A. Xiao X. Ward A.J. Castle J.C. Johnson J.M. Burge C.B. Cooper T.A. A postnatal switch of CELF and MBNL proteins reprograms alternative splicing in the developing heart.Proc. Natl. Acad. Sci. USA. 2008; 105: 20333-20338Crossref PubMed Scopus (367) Google Scholar, 8Timchenko N.A. Cai Z.J. Welm A.L. Reddy S. Ashizawa T. Timchenko L.T. RNA CUG repeats sequester CUGBP1 and alter protein levels and activity of CUGBP1.J. Biol. Chem. 2001; 276: 7820-7826Crossref PubMed Scopus (251) Google Scholar, 9Paul S. Dansithong W. Kim D. Rossi J. Webster N.J. Comai L. Reddy S. Interaction of muscleblind, CUG-BP1 and hnRNP H proteins in DM1-associated aberrant IR splicing.EMBO J. 2006; 25: 4271-4283Crossref PubMed Scopus (119) Google Scholar, 10Mankodi A. Lin X. Blaxall B.C. Swanson M.S. Thornton C.A. Nuclear RNA foci in the heart in myotonic dystrophy.Circ. Res. 2005; 97: 1152-1155Crossref PubMed Scopus (89) Google Scholar, 11Kuyumcu-Martinez N.M. Wang G.S. Cooper T.A. Increased steady-state levels of CUGBP1 in myotonic dystrophy 1 are due to PKC-mediated hyperphosphorylation.Mol. Cell. 2007; 28: 68-78Abstract Full Text Full Text PDF PubMed Scopus (350) Google Scholar The deregulated expression and activity of these RNA-binding proteins in individuals affected by DM1 leads to perturbations in the alternative splicing of key genes.7Kalsotra A. Xiao X. Ward A.J. Castle J.C. Johnson J.M. Burge C.B. Cooper T.A. A postnatal switch of CELF and MBNL proteins reprograms alternative splicing in the developing heart.Proc. Natl. Acad. Sci. USA. 2008; 105: 20333-20338Crossref PubMed Scopus (367) Google Scholar, 12Kuyumcu-Martinez N.M. Cooper T.A. Misregulation of alternative splicing causes pathogenesis in myotonic dystrophy.Prog. Mol. Subcell. Biol. 2006; 44: 133-159Crossref PubMed Scopus (62) Google Scholar The extent of the contribution of spliceopathy to CDM1 pathogenesis, however, remains unclear. Transgenic mouse models have been developed to investigate DM1 etiology.13Mankodi A. Logigian E. Callahan L. McClain C. White R. Henderson D. Krym M. Thornton C.A. Myotonic dystrophy in transgenic mice expressing an expanded CUG repeat.Science. 2000; 289: 1769-1773Crossref PubMed Scopus (566) Google Scholar, 14Ho T.H. Bundman D. Armstrong D.L. Cooper T.A. Transgenic mice expressing CUG-BP1 reproduce splicing mis-regulation observed in myotonic dystrophy.Hum. Mol. Genet. 2005; 14: 1539-1547Crossref PubMed Scopus (198) Google Scholar, 15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar Among these models, the HSALR mouse expressing 250 CTG repeats under the human skeletal actin promoter13Mankodi A. Logigian E. Callahan L. McClain C. White R. Henderson D. Krym M. Thornton C.A. Myotonic dystrophy in transgenic mice expressing an expanded CUG repeat.Science. 2000; 289: 1769-1773Crossref PubMed Scopus (566) Google Scholar and the DMSXL mouse, which harbors a mutant form of the human DMPK gene that carries >1,000 CTG repeats and is under the regulation of its own promoter. Homozygous DMSXL mice display several phenotypical manifestations of the disease and provide the only available DM1 model for screening of ASOs targeting DMPK regions outside the repeats.15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar, 16Hernández-Hernández O. Guiraud-Dogan C. Sicot G. Huguet A. Luilier S. Steidl E. Saenger S. Marciniak E. Obriot H. Chevarin C. et al.Myotonic dystrophy CTG expansion affects synaptic vesicle proteins, neurotransmission and mouse behaviour.Brain. 2013; 136: 957-970Crossref PubMed Scopus (56) Google Scholar It was previously shown that a steric blocking ASO targeting the CUGexp repeats in HSALR mice could reverse a subset of mis-spliced RNAs.17Wheeler T.M. Sobczak K. Lueck J.D. Osborne R.J. Lin X. Dirksen R.T. Thornton C.A. Reversal of RNA dominance by displacement of protein sequestered on triplet repeat RNA.Science. 2009; 325: 336-339Crossref PubMed Scopus (318) Google Scholar Using the ASO-induced RNA cleavage strategy, the same group showed that targeting the chimeric actin gene in HSALR mice could correct myotonia and also that targeting the DMPK gene was possible following systemic injections in asymptomatic DM1 mice.18Wheeler T.M. Leger A.J. Pandey S.K. MacLeod A.R. Nakamori M. Cheng S.H. Wentworth B.M. Bennett C.F. Thornton C.A. Targeting nuclear RNA for in vivo correction of myotonic dystrophy.Nature. 2012; 488: 111-115Crossref PubMed Scopus (384) Google Scholar We hypothesized the reduction in the level of mutant transcripts by targeting the DMPK transcript with an ASO-inducing RNA cleavage would alleviate pathological phenotypic traits in a mouse model of DM1, such as the loss of strength. We recently identified two ASOs, a 2′-4′-constrained ethyl (cEt) and a 2′-O-methoxyethyl (MOE) gapmers, which were able to achieve strong knockdown of DMPK mutant transcripts in DM1 cells and in a mouse model of DM1.18Wheeler T.M. Leger A.J. Pandey S.K. MacLeod A.R. Nakamori M. Cheng S.H. Wentworth B.M. Bennett C.F. Thornton C.A. Targeting nuclear RNA for in vivo correction of myotonic dystrophy.Nature. 2012; 488: 111-115Crossref PubMed Scopus (384) Google Scholar, 19Pandey S.K. Wheeler T.M. Justice S.L. Kim A. Younis H.S. Gattis D. Jauvin D. Puymirat J. Swayze E.E. Freier S.M. et al.Identification and characterization of modified antisense oligonucleotides targeting DMPK in mice and nonhuman primates for the treatment of myotonic dystrophy type 1.J. Pharmacol. Exp. Ther. 2015; 355: 329-340Crossref PubMed Scopus (92) Google Scholar Over 3,000 ASOs containing either MOE, cEt, or a combination of both chemistries were previously screened for inhibition of human DMPK expression.19Pandey S.K. Wheeler T.M. Justice S.L. Kim A. Younis H.S. Gattis D. Jauvin D. Puymirat J. Swayze E.E. Freier S.M. et al.Identification and characterization of modified antisense oligonucleotides targeting DMPK in mice and nonhuman primates for the treatment of myotonic dystrophy type 1.J. Pharmacol. Exp. Ther. 2015; 355: 329-340Crossref PubMed Scopus (92) Google Scholar We identified two ASOs, the first bearing MOE modifications complementary to a region in exon 15 upstream to the (CUG)n repeats within the 3′ UTR of DMPK, and the second bearing cEt modifications complementary to a region in exon 15 downstream to the aforementioned (CUG)n repeat, which exhibited particularly high potency (Figure 1A). ISIS 445569 and ISIS 486178 are 100% complementary to human DMPK gene; however, ISIS 486178 is also 100% complementary to the monkey and mouse Dmpk gene. Human DM1 muscle satellite cells were transfected with different concentrations (1, 5, 10, 20, 50, 125, 250, 500, and 1,000 nM) of each oligonucleotide in the presence of Lipofectamine 2000 transfection reagent (Invitrogen). Blind analysis for RNA foci was performed after 2 days of differentiation. Quantification of nuclear foci, as identified by fluorescence in situ hybridization (FISH), revealed a dose-dependent disappearance of nuclear foci (Figure 1B) in human DM1 myotubes, with a maximum effect (>90% of foci disappearance; Figure 1C) occurring at 20 nM for both ASOs (Figure S1). Northern blot analysis confirmed that both ASOs induced a 90% reduction of the mutant transcripts (Figures 1D and 1E). As expected, because both ASOs were not specific for the mutant transcripts, a 70% reduction of the wild-type DMPK transcripts was also observed. Nuclear redistribution of MBNL1 also occurred after treatment with both ASOs (Figure S2), indicating that CUGexp repeats are effectively degraded after cleavage events at either the 5′ or 3′ side of the repeat tract at the site targeted by the respective ASOs. In a previous study, we identified a set of mis-spliced RNAs present in human DM1 muscle satellite cell cultures20Klinck R. Fourrier A. Thibault P. Toutant J. Durand M. Lapointe E. Caillet-Boudin M.L. Sergeant N. Gourdon G. Meola G. et al.RBFOX1 cooperates with MBNL1 to control splicing in muscle, including events altered in myotonic dystrophy type 1.PLoS ONE. 2014; 9: e107324Crossref PubMed Scopus (32) Google Scholar that can be used to monitor alternative splicing corrections 48 hr after transfection. Treatment of DM1 cells with both ASOs consistently corrected the mis-splicing of SORBS1, CAMK2G, DMD, PDLIM3, and TTN transcripts (Figure 1F). In order to assess the potential cellular toxicity of the two ASOs under study, a microarray was performed in normal myotubes treated with these ASOs at ten times their optimal concentration. Using gene ontology (GO) assignments to categorize genes for their involvement in apoptosis, cytotoxicity, and inflammation processes, no genes assigned to these GO categories were significantly affected (i.e., by a change ≥2-fold). Other genes affected by ISIS 486178 are listed in Table S1. Moreover, we performed a cell apoptosis and necrosis assay (Figure S3) and were unable to detect any significant cell-death-associated event. Therefore, ISIS 445569 and ISIS 486178 appeared to display efficacy and tolerability in cell culture, supporting advancing into in vivo studies. On the basis of previous data, experimental procedures were done in homozygous female mice rather than in males because muscle function analysis was more consistent.15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar We evaluated these two ASOs in homozygous DMSXL mice by subcutaneous (s.c.) injection of 25 mg/kg (ISIS 486178) and 50 mg/kg (ISIS 445569) of ASO biweekly for the first 4 weeks, and then once a week for the following 5 weeks. Dosage was chosen based on dose-response experiments previously performed in heterozygous DMSXL mice.19Pandey S.K. Wheeler T.M. Justice S.L. Kim A. Younis H.S. Gattis D. Jauvin D. Puymirat J. Swayze E.E. Freier S.M. et al.Identification and characterization of modified antisense oligonucleotides targeting DMPK in mice and nonhuman primates for the treatment of myotonic dystrophy type 1.J. Pharmacol. Exp. Ther. 2015; 355: 329-340Crossref PubMed Scopus (92) Google Scholar After 9 weeks of administration (13 injections), ASOs reduced the number of nuclear foci by 70% and 40% in the quadriceps (Figures 2A and S4) for ISIS 486178 and ISIS 445569, respectively. Furthermore, RT-qPCR confirmed these numbers, with an average of 66% (ISIS 486178) and 41% (ISIS 445569) reduction in human mutant DM1 transcripts in six skeletal muscles (tibialis anterior, soleus, quadriceps, latissimus dorsi, triceps, and diaphragm) (Figure 2B). Only ISIS 486178 had an effect in the heart, with a 31% decrease in mutant transcripts, and no significant decrease was observed in the brain with either ASO, as expected based on poor blood-brain barrier penetration of ASOs.21Geary R.S. Yu R.Z. Watanabe T. Henry S.P. Hardee G.E. Chappell A. Matson J. Sasmor H. Cummins L. Levin A.A. Pharmacokinetics of a tumor necrosis factor-alpha phosphorothioate 2′-O-(2-methoxyethyl) modified antisense oligonucleotide: comparison across species.Drug Metab. Dispos. 2003; 31: 1419-1428Crossref PubMed Scopus (143) Google Scholar These data indicate that the cEt-containing DMPK ASO ISIS 486178 is more efficient than the MOE ASO ISIS 445569 for targeting mutant transcripts in muscle tissues after systemic injection. We have previously shown that homozygous DMSXL female mice have a reduced body weight compared to age-matched wild-type mice.15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar To determine whether the decrease in CUGexp RNAs by ASOs attenuate this loss in body weight, DMSXL mice were treated with 25 mg/kg of ISIS 486178 twice a week for 4 weeks. A significant increase in body weight was observed after 4 weeks of treatment (p = 0.016). On the other hand, no significant effect was observed with the less potent ASO ISIS 445569 (Table 1).Table 1Body Weight in Female DMSXL Mice Treated for 2 Months with ASOsMean Body Weight (g) ± SDDay 0Day 30Day 60Δ60p ValueDMSXL18.1 ± 1.918.4 ± 1.818.2 ± 2.40.10.5ISIS 48617817.2 ± 1.918.3 ± 1.618.7 ± 1.71.50.016ISIS 44556917.4 ± 1.518.2 ± 2.018.4 ± 3.11.00.2Wild-type22.8 ± 1.623.9 ± 1.223.8 ± 1.71.70.06(n = 10; ISIS 445569, n = 8) p values by unpaired two-tailed t test. Open table in a new tab (n = 10; ISIS 445569, n = 8) p values by unpaired two-tailed t test. Ideally, a therapeutic reduction of CUGexp RNAs would lead to increased muscle strength in the DMSXL mice. We have previously shown that these mice exhibit a 30% reduction in forelimb strength compared to age-matched wild-type mice using the grip test.15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar During the 2-month treatment period, untreated homozygous DMSXL mice lost 5.9 g in force, whereas treatment with ASO ISIS 486178 significantly increased muscle strength by 2.6 g (p = 0.003). A partial improvement of 0.5 g was seen following ISIS 445569 treatment, but was not significant when compared to average force variation in the DXSXL mice (Figure 2C). As in a previous characterization of these mice,15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar the correlation between strength phenotype and specific force was not possible (data not shown). Studies on rat soleus muscle development revealed an early muscle fiber distribution, with mixed fibers type 1/2a (2c) transitioning to type 1 after a few months of life.22Ansved T. Larsson L. Effects of ageing on enzyme-histochemical, morphometrical and contractile properties of the soleus muscle in the rat.J. Neurol. Sci. 1989; 93: 105-124Abstract Full Text PDF PubMed Scopus (71) Google Scholar Four-month-old DMSXL mice have been reported to exhibit a higher proportion of fibers 1/2a in soleus muscle than have age-matched wild-type mice,15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar which might be linked to delayed muscle fiber differentiation, as found in CDM1 patients.2Farkas-Bargeton E. Barbet J.P. Dancea S. Wehrle R. Checouri A. Dulac O. Immaturity of muscle fibers in the congenital form of myotonic dystrophy: its consequences and its origin.J. Neurol. Sci. 1988; 83: 145-159Abstract Full Text PDF PubMed Scopus (66) Google Scholar This prompted us to investigate the possibility of reversing the persistent fiber immaturity profile following treatment with ASOs. Immunostaining of soleus muscle fibers, which express both myosin heavy chain 1 and 2a in 7-month-old homozygous DMSXL revealed a 2.7% abundance of type 1/2a fibers, whereas only 0.85% of these fibers can be observed in age-matched wild-type mice. Treatment of DMSXL mice with ASO ISIS 486178 restored 95% of the normal profile of type 1/2a fibers in DMSXL mice (Figure 2E). Treatment of DMSXL mice with ISIS 486178 and ISIS 445569 for 9 weeks was not associated with any mortality. One untreated DMSXL mouse died a few hours prior to necropsy, and another died before the first injection. No DMSXL mice treated with ASO ISIS 486178 and ISIS 445569 died during the experiment. No liver anomalies were visually detected during necropsy, whereas alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate transaminase (AST) blood levels all remained within normal limits, without any significant variation (Figure 2D). Likewise, creatine kinase (CPK) and creatinine (CRE) levels, which are markers for tissue and kidney damage, respectively, were shown to stay within the normal thresholds for the course of the experiment. A microarray on tibialis anterior muscle samples from ISIS-486178-treated DMSXL mice provided a good opportunity to examine its effect on the expression inflammation, necrosis, apoptosis, and other cell death genes using GO analysis. Only five endogenous genes were found to be downregulated by treatment (i.e., by ≥2-fold change; Table S2), namely Ibsp, Atp2a2, Meg3, Nnat, and Tpm3 (Figure 2F). The microarray also corroborated our RT-qPCR results for Dmpk gene expression, with a 73% reduction. Because the ISIS 486178 sequence also targets the endogenous mouse Dmpk transcripts, we next analyzed its effect in wild-type mice following s.c. injection. After 9 weeks of administration, a strong decrease in endogenous Dmpk mRNA was detected (Figure S5). Despite the strong downregulation of Dmpk expression, no significant changes in growth, mortality, or muscle strength were detectable after treatment (data not shown). These results indicate that the decrease in endogenous Dmpk mRNA by ISIS 486178 had no effect on the DM1 phenotype, in good agreement with our previous data19Pandey S.K. Wheeler T.M. Justice S.L. Kim A. Younis H.S. Gattis D. Jauvin D. Puymirat J. Swayze E.E. Freier S.M. et al.Identification and characterization of modified antisense oligonucleotides targeting DMPK in mice and nonhuman primates for the treatment of myotonic dystrophy type 1.J. Pharmacol. Exp. Ther. 2015; 355: 329-340Crossref PubMed Scopus (92) Google Scholar and with data showing that knockout of the Dmpk gene in transgenic mice induce only a late-onset progressive myopathy.23Reddy S. Smith D.B. Rich M.M. Leferovich J.M. Reilly P. Davis B.M. Tran K. Rayburn H. Bronson R. Cros D. et al.Mice lacking the myotonic dystrophy protein kinase develop a late onset progressive myopathy.Nat. Genet. 1996; 13: 325-335Crossref PubMed Scopus (288) Google Scholar The latter observations suggest that the residual Dmpk mRNA levels might be sufficient to counteract these effects and sustain normal function. Homozygous DMSXL mice display high mortality during the first month of life, as well as growth retardation, decrease in muscle strength, motor performance, and cognitive impairments, but no or only mild myotonia.16Hernández-Hernández O. Guiraud-Dogan C. Sicot G. Huguet A. Luilier S. Steidl E. Saenger S. Marciniak E. Obriot H. Chevarin C. et al.Myotonic dystrophy CTG expansion affects synaptic vesicle proteins, neurotransmission and mouse behaviour.Brain. 2013; 136: 957-970Crossref PubMed Scopus (56) Google Scholar Histological features of the skeletal muscles revealed a decrease in the mean muscle fiber cross-sectional area, an increase in the number of oxidative fibers, and an increase in mixed fibers 1/2a, which may point to a potential delay in muscle maturation. The clinical and histological manifestations observed in DMSXL mice are characteristically closer to those observed in the congenital rather than the adult form of the disease. The absence of the classical histological features observed in human DM1 skeletal muscle, such as type 1 fiber atrophy and type 2 fiber hypertrophy, ringed fibers, sarcoplasmic mass, and mild splicing abnormalities are also consistent with this observation.1Harper P.S. Myotonic Dystrophy. Saunders, 2001Google Scholar There is indeed a lack of data for the involvement of the spliceopathy in the pathogenesis of the congenital form of the disease. Very mild and variable mis-splicings have been characterized in DMSXL mice15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar but did not prove to represent a significant measure of pathological outcome in this model for ASO screening. Insertional effects in DMSXL mice can be ruled out because parental homozygous mice carrying 500–600 repeats (DM300) have very mild or no detectable phenotype.15Huguet A. Medja F. Nicole A. Vignaud A. Guiraud-Dogan C. Ferry A. Decostre V. Hogrel J.Y. Metzger F. Hoeflich A. et al.Molecular, physiological, and motor performance defects in DMSXL mice carrying >1,000 CTG repeats from the human DM1 locus.PLoS Genet. 2012; 8: e1003043Crossref PubMed Scopus (87) Google Scholar, 24Seznec H. Agbulut O. Sergeant N. Savouret C. Ghestem A. Tabti N. Willer J.C. Ourth L. Duros C. Brisson E. et al.Mice transgenic for the human myotonic dystrophy region with expanded CTG repeats display muscular and brain abnormalities.Hum. Mol. Genet. 2001; 10: 2717-2726Crossref PubMed Scopus (194) Google Scholar, 25Panaite P.A. Kielar M. Kraftsik R. Gourdon G. Kuntzer T. Barakat-Walter I. Peripheral neuropathy is linked to a severe form of myotonic dystrophy in transgenic mice.J. Neuropathol. Exp. Neurol. 2011; 70: 678-685Crossref PubMed Scopus (14) Google Scholar Short-term therapy with ASOs produced a sustained phenotypic disease reversal, including gain of body weight, increase in muscle strength, and restoration in skeletal muscle maturation. These results show that muscle weakness in DM1 can be improved using a targeted approach to eliminate toxic RNAs. This indicates that muscle weakness may be reversible in a DM1 patient. In this report, we demonstrate that a cEt-modified ASO (ISIS 486178) targeting DMPK mRNA significantly reduced mutant transcripts in cells and generates a reproducible and robust reduction in skeletal muscle DMPK mRNAs in DM1 mice following s.c. administration. The MOE gapmer ASOs have been shown to reduce mutant DMPK transcripts in the skeletal muscles of DMSXL mice following s.c. administration, but at higher doses than those required for the cEt-modified ASOs. A 50% and 75% reduction in DMPK RNAs were observed in skeletal muscles at 50 and 75 mg/kg, respectively, twice a week for 4 weeks using MOE gapmer ASOs.18Wheeler T.M. Leger A.J. Pandey S.K. MacLeod A.R. Nakamori M. Cheng S.H. Wentworth B.M. Bennett C.F. Thornton C.A. Targeting nuclear RNA for in vivo correction of myotonic dystrophy.Nature. 2012; 488: 111-115Crossref PubMed Scopus (384) Google Scholar The potent activity of cEt-modified oligonucleotides in skeletal muscle has also been demonstrated for an androgen receptor (AR) targeting ASO in AR transgenic mice.26Lieberman A.P. Yu Z. Murray S. Peralta R. Low A. Guo S. Yu X.X. Cortes C.J. Bennett C.F. Monia B.P. et al.Peripheral androgen receptor gene suppression rescues disease in mouse models of spinal and bulbar muscular atrophy.Cell Rep. 2014; 7: 774-784Abstract Full Text Full Text PDF PubMed Scopus (118) Google Scholar Activity in skeletal muscle has also been described for a peptide-PMO conjugate, Pip5e-PMO, with a single dose of 12–25 mg/kg and showed highly efficient exon skipping and dystrophin production in mdx mice, with >80% dystrophin-positive fibers measured in all skeletal muscle groups after systemic injection.27Yin H. Saleh A.F. Betts C. Camelliti P. Seow Y. Ashraf S. Arzumanov A. Hammond S. Merritt T. Gait M.J. et al.Pip5 transduction peptides direct high efficiency oligonucleotide-mediated dystrophin exon skipping in heart and phenotypic correction in mdx mice.Mol. Ther. 2011; 19: 1295-1303Abstract Full Text Full Text PDF PubMed Scopus (102) Google Scholar In contrast with what was observed with the cEt-modified ASO in DMSXL mice, no phenotypic reversion was observed following systemic injection of 12 mg/kg Pip6a-PMO (more efficient peptide ASO than Pip5e-PMO) likely due to uncompromised muscle cell membranes in DMSXL mice (J. Puymirat, unpublished data). The difference between the dose of ASOs used in mdx and DMSXL mice may be explained by more than one factor. The physical chemical behavior of the neutral PMO to the negatively charged cEt ASO will affect cell intake. Second, there is the matter of the cell-penetrating peptide attached to the PMO, which may improve cell penetration. Lastly, it has been shown that the muscle membrane is more permeable in mdx but not DMSXL mice,28González-Barriga A. Kranzen J. Croes H.J. Bijl S. van den Broek W.J. van Kessel I.D. van Engelen B.G. van Deutekom J.C. Wieringa B. Mulders S.A. et al.Ce" @default.
• W2616392786 created "2017-05-26" @default.
• W2616392786 creator A5016432689 @default.
• W2616392786 creator A5026191697 @default.
• W2616392786 creator A5036673188 @default.
• W2616392786 creator A5037948824 @default.
• W2616392786 creator A5039206363 @default.
• W2616392786 creator A5043092391 @default.
• W2616392786 creator A5047626547 @default.
• W2616392786 creator A5056021094 @default.
• W2616392786 creator A5058301400 @default.
• W2616392786 creator A5070676987 @default.
• W2616392786 creator A5083296071 @default.
• W2616392786 creator A5086348646 @default.
• W2616392786 creator A5087711568 @default.
• W2616392786 date "2017-06-01" @default.
• W2616392786 modified "2023-10-18" @default.
• W2616392786 title "Targeting DMPK with Antisense Oligonucleotide Improves Muscle Strength in Myotonic Dystrophy Type 1 Mice" @default.
• W2616392786 cites W1212882679 @default.
• W2616392786 cites W1510644906 @default.
• W2616392786 cites W162861282 @default.
• W2616392786 cites W1738068946 @default.
• W2616392786 cites W1879316584 @default.
• W2616392786 cites W1955360692 @default.
• W2616392786 cites W1964514885 @default.
• W2616392786 cites W1968949098 @default.
• W2616392786 cites W1969986482 @default.
• W2616392786 cites W1975664052 @default.
• W2616392786 cites W1992481904 @default.
• W2616392786 cites W1992609622 @default.
• W2616392786 cites W1999613141 @default.
• W2616392786 cites W2004824383 @default.
• W2616392786 cites W2008897130 @default.
• W2616392786 cites W2017774656 @default.
• W2616392786 cites W2030823729 @default.
• W2616392786 cites W2036075451 @default.
• W2616392786 cites W2039944783 @default.
• W2616392786 cites W2050679356 @default.
• W2616392786 cites W2059459126 @default.
• W2616392786 cites W2078217107 @default.
• W2616392786 cites W2079759083 @default.
• W2616392786 cites W2081098333 @default.
• W2616392786 cites W2081748748 @default.
• W2616392786 cites W2084091637 @default.
• W2616392786 cites W2091704340 @default.
• W2616392786 cites W2099039583 @default.
• W2616392786 cites W2099191309 @default.
• W2616392786 cites W2109385021 @default.
• W2616392786 cites W2110640321 @default.
• W2616392786 cites W2112137134 @default.
• W2616392786 cites W2122776931 @default.
• W2616392786 cites W2123557978 @default.
• W2616392786 cites W2131447660 @default.
• W2616392786 cites W2143228863 @default.
• W2616392786 cites W2145025818 @default.
• W2616392786 cites W2153583721 @default.
• W2616392786 cites W2158442258 @default.
• W2616392786 cites W2165510835 @default.
• W2616392786 cites W2170989872 @default.
• W2616392786 cites W2239874339 @default.
• W2616392786 cites W2587419486 @default.
• W2616392786 cites W4211120586 @default.
• W2616392786 doi "https://doi.org/10.1016/j.omtn.2017.05.007" @default.
• W2616392786 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5453865" @default.
• W2616392786 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/28624222" @default.
• W2616392786 hasPublicationYear "2017" @default.
• W2616392786 type Work @default.
• W2616392786 sameAs 2616392786 @default.
• W2616392786 citedByCount "67" @default.
• W2616392786 countsByYear W26163927862018 @default.
• W2616392786 countsByYear W26163927862019 @default.
• W2616392786 countsByYear W26163927862020 @default.
• W2616392786 countsByYear W26163927862021 @default.
• W2616392786 countsByYear W26163927862022 @default.
• W2616392786 countsByYear W26163927862023 @default.
• W2616392786 crossrefType "journal-article" @default.
• W2616392786 hasAuthorship W2616392786A5016432689 @default.
• W2616392786 hasAuthorship W2616392786A5026191697 @default.
• W2616392786 hasAuthorship W2616392786A5036673188 @default.
• W2616392786 hasAuthorship W2616392786A5037948824 @default.
• W2616392786 hasAuthorship W2616392786A5039206363 @default.
• W2616392786 hasAuthorship W2616392786A5043092391 @default.
• W2616392786 hasAuthorship W2616392786A5047626547 @default.
• W2616392786 hasAuthorship W2616392786A5056021094 @default.
• W2616392786 hasAuthorship W2616392786A5058301400 @default.
• W2616392786 hasAuthorship W2616392786A5070676987 @default.
• W2616392786 hasAuthorship W2616392786A5083296071 @default.
• W2616392786 hasAuthorship W2616392786A5086348646 @default.
• W2616392786 hasAuthorship W2616392786A5087711568 @default.
• W2616392786 hasBestOaLocation W26163927861 @default.
• W2616392786 hasConcept C104317684 @default.
• W2616392786 hasConcept C126322002 @default.
• W2616392786 hasConcept C129312508 @default.
• W2616392786 hasConcept C134018914 @default.
• W2616392786 hasConcept C2778063965 @default.
• W2616392786 hasConcept C2779030066 @default.
• W2616392786 hasConcept C2779959927 @default.
• W2616392786 hasConcept C2780304057 @default.

• next