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

标题：DMD的反义疗法-AON用于外显子45跳跃的优化和毒理学。 杜氏肌营养不良症（DMD）是最常见的形式的肌营养不良症影响1每3500 活产男婴。这种疾病的特点是严重的肌肉萎缩和虚弱，这在临床上变得 在3到5岁之间很明显。DMD是由肌营养不良蛋白中所谓的移码突变引起的 基因导致过早终止，没有蛋白质合成，而BMD是由突变引起的， 产生截短的，但与部分或几乎全部功能的蛋白质在框内的转录物。反义 寡核苷酸（AON）介导的外显子剪接（反义疗法）已被证明是有效的， 恢复由导致DMD的突变破坏的开放阅读框架。P.I.项目建立 事实上，肌营养不良蛋白的接近正常水平可以恢复和维持在全身骨骼肌 通过在动物模型中定期给予肽标记的吗啉代AON（PPMO）， 毒性我们还讨论了成功应用该疗法的几个重要问题， 诊所。可靠的AON体外筛选系统的开发使我们能够建立高效的AON筛选系统。 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， 临床试验时毒性最小，在这个项目中，我们的目标是将成功的 实验治疗和临床治疗DMD。实验计划以实现明确的 里程碑：1）选择最有效的AON用于靶向人肌营养不良蛋白外显子45，其去除可以 治疗大部分DMD患者。这一目标将通过建立细胞培养系统来实现。（二） 确定所选PPMO在动物模型中的功效以及所选PPMO在细胞中的脱靶效应。 cultures. 3)进行毒理学测试，从而可以向FDA提交研究性新药申请。 反义治疗对DMD动物模型的治疗是有效的，并为治疗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.