Faculty of Medicine, Department of Cellular and Molecular Medicine and Centre for Neuromuscular Disease, University of Ottawa, ON, Canada K1H 8M5.
Hum Mol Genet. 2013 Aug 1;22(15):3093-111. doi: 10.1093/hmg/ddt165. Epub 2013 Apr 10.
Several therapeutic approaches are currently being developed for Duchenne muscular dystrophy (DMD) including upregulating the levels of endogenous utrophin A in dystrophic fibers. Here, we examined the role of post-transcriptional mechanisms in controlling utrophin A expression in skeletal muscle. We show that activation of p38 leads to an increase in utrophin A independently of a transcriptional induction. Rather, p38 controls the levels of utrophin A mRNA by extending the half-life of transcripts via AU-rich elements (AREs). This mechanism critically depends on a decrease in the functional availability of KSRP, an RNA-binding protein known to promote decay of ARE-containing transcripts. In vitro and in vivo binding studies revealed that KSRP interacts with specific AREs located within the utrophin A 3' UTR. Electroporation experiments to knockdown KSRP led to an increase in utrophin A in wild-type and mdx mouse muscles. In pre-clinical studies, treatment of mdx mice with heparin, an activator of p38, causes a pronounced increase in utrophin A in diaphragm muscle fibers. Together, these studies identify a pathway that culminates in the post-transcriptional regulation of utrophin A through increases in mRNA stability. Furthermore, our results constitute proof-of-principle showing that pharmacological activation of p38 may prove beneficial as a novel therapeutic approach for DMD.
目前正在开发几种治疗杜氏肌营养不良症 (DMD) 的方法,包括上调营养不良纤维中内源性 utrophin A 的水平。在这里,我们研究了转录后机制在控制骨骼肌中 utrophin A 表达中的作用。我们表明,p38 的激活可独立于转录诱导而增加 utrophin A。相反,p38 通过富含 AU 的元件 (ARE) 延长转录物的半衰期来控制 utrophin A mRNA 的水平。该机制严重依赖于 KSRP 的功能可用性降低,KSRP 是一种已知可促进 ARE 包含的转录物降解的 RNA 结合蛋白。体外和体内结合研究表明,KSRP 与 utrophin A 3'UTR 内的特定 ARE 相互作用。用于敲低 KSRP 的电穿孔实验导致野生型和 mdx 小鼠肌肉中的 utrophin A 增加。在临床前研究中,用肝素(p38 的激活剂)处理 mdx 小鼠会导致膈肌纤维中 utrophin A 明显增加。总之,这些研究确定了一条途径,通过增加 mRNA 稳定性来实现 utrophin A 的转录后调节。此外,我们的结果证明了原理证明,表明 p38 的药理学激活可能作为 DMD 的一种新的治疗方法是有益的。