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Second-generation compound for the modulation of utrophin in the therapy of DMD.

Guiraud S, Squire SE, Edwards B, Chen H, Burns DT, Shah N, Babbs A, Davies SG, Wynne GM, Russell AJ, Elsey D, Wilson FX, Tinsley JM, Davies KE - Hum. Mol. Genet. (2015)

Bottom Line: These studies in the mdx mouse demonstrate that oral administration of SMT022357 leads to increased utrophin expression in skeletal, respiratory and cardiac muscles.This results in improved sarcolemmal stability and prevents dystrophic pathology through a significant reduction of regeneration, necrosis and fibrosis.This detailed evaluation of the SMT C1100 drug series strongly endorses the therapeutic potential of utrophin modulation as a disease modifying therapeutic strategy for all DMD patients irrespective of their dystrophin mutation.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT, UK, simon.guiraud@dpag.ox.ac.uk kay.davies@dpag.ox.ac.uk.

No MeSH data available.


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In vitro activity of SMT022357. (A) SMT022357 in vitro dose response in murine H2k-mdx utrnA-luc cells expressing the human utrophin promoter linked to a firefly luciferase reporter gene. Cells were treated with compound for 24 h in standard growth medium containing 1% DMSO. The chart shows relative luminescence (RLU) in relation to five different doses (0.1, 0.3, 1, 3 and 10 µM) of SMT022357. (B) SMT022357 significantly increased mRNA copy number of the utrophin transcript in murine myoblast cells. Cells were exposed to different dose of SMT022357 in standard media with 0.1% DMSO (vehicle) for 24 hours with six biological replicates. Utrophin transcripts were normalised with 18s and b2m. Values are mean ± SEM of n = 6 per condition; *P < 0.05, **P < 0.01, ***P < 0.001. (C) Relative utrophin protein expression in murine cells treated with SMT022357 was determined by western blot and standardized for α-actinin loading. Western blots showing 2.5-fold increase of utrophin protein expression with 10 µM of SMT022357 when compared with vehicle. Relative utrophin expression is shown as mean ± SEM of n = 3 per condition.
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DDV154F1: In vitro activity of SMT022357. (A) SMT022357 in vitro dose response in murine H2k-mdx utrnA-luc cells expressing the human utrophin promoter linked to a firefly luciferase reporter gene. Cells were treated with compound for 24 h in standard growth medium containing 1% DMSO. The chart shows relative luminescence (RLU) in relation to five different doses (0.1, 0.3, 1, 3 and 10 µM) of SMT022357. (B) SMT022357 significantly increased mRNA copy number of the utrophin transcript in murine myoblast cells. Cells were exposed to different dose of SMT022357 in standard media with 0.1% DMSO (vehicle) for 24 hours with six biological replicates. Utrophin transcripts were normalised with 18s and b2m. Values are mean ± SEM of n = 6 per condition; *P < 0.05, **P < 0.01, ***P < 0.001. (C) Relative utrophin protein expression in murine cells treated with SMT022357 was determined by western blot and standardized for α-actinin loading. Western blots showing 2.5-fold increase of utrophin protein expression with 10 µM of SMT022357 when compared with vehicle. Relative utrophin expression is shown as mean ± SEM of n = 3 per condition.

Mentions: SMT022357 shows a maximal increase of 3-fold in luciferase compared with vehicle (Fig. 1A). No stabilization or inhibition of the luciferase activity was noted (data not shown). Furthermore, no change in proliferation was observed after a treatment with SMT022357 (Supplementary Material, Fig. S1). In vitro dosing of murine myoblasts with 3 µm of SMT022357 led to a 50% increase in utrophin mRNA when compared with vehicle after 2 days of treatment (Fig. 1B). Treatment of murine DMD cells with SMT022357 showed a 2.5-fold increase in utrophin protein level at an optimal concentration of 10 µm after 3 days of treatment (Fig. 1C). In comparison, SMT C1100 led to a 30% increase in Utrn mRNA level and resulted in a 2.0-fold increase in UTRN protein level (41). These data demonstrate the in vitro potential of SMT022357 to increase the level of utrophin expression.Figure 1.


Second-generation compound for the modulation of utrophin in the therapy of DMD.

Guiraud S, Squire SE, Edwards B, Chen H, Burns DT, Shah N, Babbs A, Davies SG, Wynne GM, Russell AJ, Elsey D, Wilson FX, Tinsley JM, Davies KE - Hum. Mol. Genet. (2015)

In vitro activity of SMT022357. (A) SMT022357 in vitro dose response in murine H2k-mdx utrnA-luc cells expressing the human utrophin promoter linked to a firefly luciferase reporter gene. Cells were treated with compound for 24 h in standard growth medium containing 1% DMSO. The chart shows relative luminescence (RLU) in relation to five different doses (0.1, 0.3, 1, 3 and 10 µM) of SMT022357. (B) SMT022357 significantly increased mRNA copy number of the utrophin transcript in murine myoblast cells. Cells were exposed to different dose of SMT022357 in standard media with 0.1% DMSO (vehicle) for 24 hours with six biological replicates. Utrophin transcripts were normalised with 18s and b2m. Values are mean ± SEM of n = 6 per condition; *P < 0.05, **P < 0.01, ***P < 0.001. (C) Relative utrophin protein expression in murine cells treated with SMT022357 was determined by western blot and standardized for α-actinin loading. Western blots showing 2.5-fold increase of utrophin protein expression with 10 µM of SMT022357 when compared with vehicle. Relative utrophin expression is shown as mean ± SEM of n = 3 per condition.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4492389&req=5

DDV154F1: In vitro activity of SMT022357. (A) SMT022357 in vitro dose response in murine H2k-mdx utrnA-luc cells expressing the human utrophin promoter linked to a firefly luciferase reporter gene. Cells were treated with compound for 24 h in standard growth medium containing 1% DMSO. The chart shows relative luminescence (RLU) in relation to five different doses (0.1, 0.3, 1, 3 and 10 µM) of SMT022357. (B) SMT022357 significantly increased mRNA copy number of the utrophin transcript in murine myoblast cells. Cells were exposed to different dose of SMT022357 in standard media with 0.1% DMSO (vehicle) for 24 hours with six biological replicates. Utrophin transcripts were normalised with 18s and b2m. Values are mean ± SEM of n = 6 per condition; *P < 0.05, **P < 0.01, ***P < 0.001. (C) Relative utrophin protein expression in murine cells treated with SMT022357 was determined by western blot and standardized for α-actinin loading. Western blots showing 2.5-fold increase of utrophin protein expression with 10 µM of SMT022357 when compared with vehicle. Relative utrophin expression is shown as mean ± SEM of n = 3 per condition.
Mentions: SMT022357 shows a maximal increase of 3-fold in luciferase compared with vehicle (Fig. 1A). No stabilization or inhibition of the luciferase activity was noted (data not shown). Furthermore, no change in proliferation was observed after a treatment with SMT022357 (Supplementary Material, Fig. S1). In vitro dosing of murine myoblasts with 3 µm of SMT022357 led to a 50% increase in utrophin mRNA when compared with vehicle after 2 days of treatment (Fig. 1B). Treatment of murine DMD cells with SMT022357 showed a 2.5-fold increase in utrophin protein level at an optimal concentration of 10 µm after 3 days of treatment (Fig. 1C). In comparison, SMT C1100 led to a 30% increase in Utrn mRNA level and resulted in a 2.0-fold increase in UTRN protein level (41). These data demonstrate the in vitro potential of SMT022357 to increase the level of utrophin expression.Figure 1.

Bottom Line: These studies in the mdx mouse demonstrate that oral administration of SMT022357 leads to increased utrophin expression in skeletal, respiratory and cardiac muscles.This results in improved sarcolemmal stability and prevents dystrophic pathology through a significant reduction of regeneration, necrosis and fibrosis.This detailed evaluation of the SMT C1100 drug series strongly endorses the therapeutic potential of utrophin modulation as a disease modifying therapeutic strategy for all DMD patients irrespective of their dystrophin mutation.

View Article: PubMed Central - PubMed

Affiliation: Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT, UK, simon.guiraud@dpag.ox.ac.uk kay.davies@dpag.ox.ac.uk.

No MeSH data available.


Related in: MedlinePlus