Duchenne Cardiomyopathy Gene Therapy

描述（由申请人提供）：心肌病是杜氏肌营养不良症（DMD）的主要死亡原因，DMD是最常见的儿童致死性肌肉疾病。DMD是由肌营养不良蛋白基因突变引起的，目前还没有治愈方法。腺相关病毒（AAV）介导的微小肌营养不良蛋白基因治疗在改善Duchenne骨骼肌疾病方面显示出巨大的前景。然而，我们最近发现，为治疗骨骼肌疾病而开发的简短基因可能无法完全满足心脏的需要。在这里，我们假设杜氏心肌病基因治疗可能需要一个特定的肌营养不良蛋白结构域，是目前可用的微/minigenes失踪。回顾过去17年来Duchenne心肌病相关的临床报告，我们确定了一个假定的心脏保护结构域的dystrophin基因。在这个提议中，我们将测试我们是否可以通过在微基因中包括推定的心脏保护结构域来实现更好的心脏拯救。具体地，将产生携带推定的心脏保护结构域的新的微小基因。AAV将用于将这些微小基因递送至杜氏心肌病小鼠模型的心脏。将使用全面的解剖、细胞、生化和生理分析来监测心脏抢救。新的微/微基因的治疗效果也将与当前的微/微基因进行比较。我们的长期目标是开发一种有效的AAV基因疗法来治疗患者。在开始人体试验之前的一个关键步骤是在犬DMD模型中进行临床前评价。我们假设AAV基因治疗可以改善金毛犬肌营养不良（GRMD）模型中的心肌病。在鼠模型中鉴定的最佳微小基因将通过系统性AAV基因转移递送至新生GRMD幼犬。正常犬和注射盐水的GRMD犬将作为对照。心脏病的进展以及基因转移效率将使用我们已经开发的解剖学，组织学，细胞学，生物化学和生理学分析的全面面板进行仔细监测。综上所述，我们的研究将显著推进Duchenne心肌病基因治疗。 公共卫生相关性：杜氏肌营养不良症（DMD）是一种致死性疾病，影响相当大的患者群体（约3,500名新生儿中有1名）。DMD相关的心脏病显著降低患者的生活质量和寿命。因此，我们建议发展腺相关病毒基因疗法来治疗杜氏心脏病。我们的发现将为最终治愈DMD铺平道路。

DESCRIPTION (provided by applicant): Cardiomyopathy is a leading cause of death in Duchenne muscular dystrophy (DMD), the most common childhood lethal muscle disease. DMD is caused by dystrophin gene mutation and there is currently no cure. Adeno-associated virus (AAV)-mediated micro/mini-dystrophin gene therapy has shown great promise in ameliorating Duchenne skeletal muscle disease. However, we recently found that the abbreviated genes that were developed for treating skeletal muscle disease may not completely fulfill the needs of the heart. Here, we hypothesize that Duchenne cardiomyopathy gene therapy may require a specific dystrophin domain that is missing in the current available micro/minigenes. On reviewing Duchenne cardiomyopathy-related clinical reports over the last 17 years, we identified a putative heart protection domain in the dystrophin gene. In this proposal, we will test whether we can achieve better cardiac rescue by including the putative heart protection domain in the micro/minigenes. Specifically, novel micro/minigenes carrying the putative heart protection domain will be generated. AAV will be used to deliver these micro/minigenes to the heart in the mouse models of Duchenne cardiomyopathy. Comprehensive anatomic, cellular, biochemical, and physiological assays will be used to monitor cardiac rescue. The therapeutic efficacy of new micro/minigenes will also be compared to that of the current micro/minigenes. Our long-term goal is to develop an effective AAV gene therapy to treat patients. A critical step before initiating human trial is preclinical evaluation in the canine DMD model. We hypothesize that AAV gene therapy can ameliorate cardiomyopathy in the golden retriever muscular dystrophy (GRMD) model. The best micro/minigenes identified in the murine model will be delivered to neonatal GRMD puppy by systemic AAV gene transfer. Normal dogs and saline injected GRMD dogs will be included as controls. Progression of the heart disease as well as gene transfer efficiency will be carefully monitored using a comprehensive panel of anatomic, histological, cellular, biochemical, and physiological assays we already developed. Taken together, our study will significantly advance Duchenne cardiomyopathy gene therapy. PUBLIC HEALTH RELEVANCE: Duchenne muscular dystrophy (DMD) is a lethal disease affecting a fairly large population of patients (~one in 3,500 newborn boys). DMD related heart disease significantly reduces the life quality and life span of patients. Here we propose to develop AAV gene therapy to treat Duchenne heart disease. Our findings will pave the way to eventually cure DMD.