Antisense therapy for DMD-Optimization and toxicology of AON for exon 45 skipping

标题：DMD的反义治疗-Aon对外显子45跳过的优化和毒理学。 Duchenne肌营养不良症(DMD)是最常见的肌营养不良症，每3500例中就有1例 活的男婴。这种疾病的特征是严重的肌肉萎缩和虚弱，这在临床上变得更加明显。 在3至5岁之间表现明显。DMD是由dystrophin中的所谓移码突变引起的 导致早产的基因没有蛋白质合成，而骨密度是由突变引起的， 用部分或几乎全部功能的蛋白质创建截短的框内转录本。反义 寡核苷酸(AON)介导的外显子剪接(反义治疗)已被证明对 因引起DMD的突变而中断的开放阅读框架的恢复。P.I.已经建立了一个项目 在全身骨骼肌中可以恢复和保持接近正常水平的抗肌营养不良蛋白 在可耐受的动物模型中常规给予多肽标记的吗啡Aon(PPMO) 毒性。我们还解决了几个重要的问题，这些问题对于成功地将该疗法应用于 诊所。建立一种可靠的体外AON筛选系统使我们能够建立高效的 AON作为靶向单个外显子的特异性药物。我们已经定义了治疗的给药方案 临床试验可达到毒性最小的效果。在这个项目中/我们的目标是将成功的 实验疗法对临床治疗DMD的作用。实验计划的中心是实现明确定义的 里程碑：1)选择最有效的针对人dystrophin外显子45的Aon，去除该外显子可以 治疗很大比例的DMD患者。这一目标将通过已建立的细胞培养系统实现。2) 确定所选PPMO在动物模型中的有效性和所选PPMO在细胞中的非靶向效应 文化。3)进行毒理学测试，从而可以向FDA提交研究性新药申请。 反义治疗在治疗DMD动物模型中是有效的，为治疗DMD提供了现实的希望。 大多数DMD。我们的目标是将这种疗法转化为临床试验。

Title: Antisense therapy for DMD-Optimization and toxicology of AON for exon 45 skipping. Duchenne muscular dystrophy (DMD) is the most common form of muscular dystrophy affecting 1 in every 3500 live male births. The disease is characterized by severe muscle wasting and weakness, which becomes clinically evident between the ages of 3 to 5 years. DMD is caused by so called frame-shift mutations in the dystrophin gene leading to premature termination with no protein synthesis whereas BMD is caused by mutations which create truncated, but in-frame transcripts with proteins of partial or nearly full function. Antisense oligonucleotide (AON)-mediated exon splicing (antisense therapy) has been shown to be effective for restoration of the open reading frame disrupted by mutations that cause DMD. The project P.I. has established the facts that near normal levels of dystrophin can be restored and maintained in the body-wide skeletal muscles through regular administration of a peptide-tagged morpholino AON (PPMO) in animal models with tolerable ¿ toxicity. We have also addressed several important issues critical for successful applications of the therapy to clinics. The development of a reliable in vitro system for AON selection enables us to establish highly effective AONs as specific drugs to target individual exons. We have defined the regimen of administration so therapeutic effect can be achieved with minimum toxicity for clinical trial. In this project/we aim to translate the successful experimental therapy to clinical treatment of DMD. The experimental plan is centered to achieve well-defined milestones: 1) Select most efficacious AON for targeting human dystrophin exon 45, the removal of which can treat a large proportion of DMD patients. This aim will be achieved with eatablished cell culture systems. 2) Determine the efficacy of selected PPMOs in animal models ahd off-target effects of the selected PPMOs in cell cultures. 3) Conduct toxicology tests, thus an investigational new drug application can be submitted to the FDA. Antisense therapy is effective in treating animal models of DMD and provides a realistic hope for treating the majority of DMD. We aim to translate the therapy into clinical trials.