Duchenne Cardiomyopathy Gene Therapy

描述(由申请人提供)：心肌病是Duchenne肌营养不良症(DMD)的主要死亡原因，Duchenne肌营养不良症是儿童最常见的致命肌肉疾病。DMD是由dystrophin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/微型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缩写基因可能不能完全满足心脏的需要。在这里，我们假设Duchenne心肌病基因治疗可能需要一个特定的dystrophin结构域，而目前可用的微/微型基因中缺少该结构域。在回顾过去17年与杜兴心肌病相关的临床报告后，我们确定了dystrophin基因中一个假定的心脏保护域。在这项提案中，我们将测试是否可以通过将假定的心脏保护结构域包括在微型/微型基因中来实现更好的心脏抢救。具体地说，将产生携带假定的心脏保护结构域的新的微型/微型基因。在Duchenne心肌病小鼠模型中，AAV将被用于将这些微型/微型基因输送到心脏。将使用综合的解剖、细胞、生化和生理分析来监测心脏抢救。新的微型/微型遗传疗法的疗效也将与目前微型/微型遗传疗法的疗效进行比较。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。在开始人类试验之前，关键的一步是对犬DMD模型进行临床前评估。我们假设AAV基因治疗可以改善金毛猎犬肌营养不良症(GRMD)模型的心肌病。在小鼠模型中确定的最好的微型/微型基因将通过系统的AAV基因转移传递给新生GRMD幼犬。正常犬和注射生理盐水的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一套全面的解剖学、组织学、细胞学、生化和生理学测试来仔细监测。综上所述，我们的研究将大大推进杜兴心肌病的基因治疗。 公共卫生相关性：杜氏肌营养不良症(DMD)是一种致命性疾病，影响相当大的患者群体(约每3500名新生儿中就有一名)。与DMD相关的心脏病显著降低了患者的生活质量和寿命。在这里，我们建议开发AAV基因疗法来治疗Duchenne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.