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，是最常见的儿童致命性肌肉疾病。DMD是由肌营养不良蛋白基因突变引起的，目前尚无治愈方法。腺相关病毒（AAV）介导的微/迷你肌营养不良蛋白基因治疗在改善杜氏骨骼肌病方面显示出巨大的希望。然而，我们最近发现，为治疗骨骼肌疾病而开发的缩短基因可能不能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，这是目前可用的微/小基因中缺失的。通过回顾过去17年杜氏心肌病相关的临床报告，我们在肌营养不良蛋白基因中发现了一个假定的心脏保护结构域。在本提案中，我们将测试是否可以通过在微/小基因中加入假定的心脏保护结构域来实现更好的心脏救援。具体来说，将产生新的携带假定心脏保护域的微/迷你基因。在杜氏心肌病小鼠模型中，AAV将用于将这些微/微小基因传递到心脏。综合解剖、细胞、生化和生理分析将用于监测心脏抢救。新的微/微基因的治疗效果也将与现有的微/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关键步骤是犬DMD模型的临床前评估。我们假设AAV基因治疗可以改善金毛肌营养不良（GRMD）模型的心肌病。在小鼠模型中鉴定出的最佳微/微小基因将通过系统AAV基因转移传递给新生GRMD幼犬。将正常犬和注射生理盐水的GRMD犬作为对照。心脏病的进展以及基因转移效率将通过我们已经开发的解剖学、组织学、细胞、生化和生理学的综合检测来仔细监测。总之，我们的研究将显著推进杜氏心肌病的基因治疗。

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.