A drug-based approach for integrin-mediated alleviation of muscular dystrophy

描述(申请人提供)：杜氏肌营养不良症(DMD)是一种严重的肌肉萎缩疾病，会导致过早死亡。目前，这种毁灭性的神经肌肉疾病还没有治愈或有效的治疗方法。在DMD患者和MDX小鼠模型的肌肉中，17整合素的表达增加。缺乏营养不良蛋白和17整合素的小鼠表现出严重的肌肉营养不良，并在4周前死亡。最近，我们证明了在骨骼肌中增强17整合素的转基因表达可以提高营养不良小鼠的生存能力。总之，这些研究表明，17整合素是DMD患者和mdx小鼠肌肉中的主要遗传修饰物，并表明上调17整合素基因表达的化合物可能成为DMD的潜在治疗方法。在这项研究中，我们将启动一个小分子发现计划，以确定针对17整合素基因表达的化合物。我们将使用从转基因小鼠中分离的克隆肌肉细胞系来开发整合素基因表达的荧光检测系统。该系统将允许在其正常的细胞和基因组环境中评估17个整合素启动子的活性。这一基于细胞的系统将用已知能增加17个整合素基因表达的化合物进行测试。然后，经过验证的基于细胞的分析将被应用于自动化的高通量药物筛选，以识别在培养的小鼠肌肉细胞中增加17个整合素基因表达的化合物。阳性化合物将在培养的小鼠和人类肌肉细胞中进行测试，以确定药物的有效性、毒性和转化为人类肌肉细胞的能力。总之，这些目标将使我们能够检验这样一个假设，即小化合物可以增加小鼠和人类肌肉细胞中整合素基因的表达。识别增加17个整合素基因表达的小化合物将为未来的研究奠定基础，这些研究将把我们的基础生物医学研究转化为未来人类临床试验的药物开发。 杜氏肌营养不良症是一种毁灭性的肌肉疾病，在美国有近5万名儿童受到影响。17整合素表达的增加已被证明对这种疾病的小鼠模型有极大的好处。本研究旨在寻找能提高17种整合素基因表达的药物，这些药物可能被证明对患有DMD的患者有治疗价值。

DESCRIPTION (provided by applicant): Duchenne Muscular Dystrophy (DMD) is a severe muscle wasting disease that leads to premature death. Currently, there is no cure or effective treatment for this devastating neuromuscular disease. Expression of the 17 integrin is increased in the muscle of DMD patients and the mdx mouse model. Mice lacking both dystrophin and the 17 integrin exhibit severe muscular dystrophy and die before 4 weeks of age. Recently we demonstrated that enhanced transgenic expression of the 17 integrin in skeletal muscle can increase the viability of dystrophic mice. Together these studies show that the 17 integrin is a major genetic modifier in the muscle of DMD patients and mdx mice and suggest that compounds that up-regulate 17 integrin gene expression may serve as a potential therapy for DMD. In this study we will initiate a small-molecule discovery program to identify compounds that target 17 integrin gene expression. We will use a clonal muscle cell line isolated from transgenic mice to develop a fluorescent assay system for integrin gene expression. This system will allow an assessment of 17 integrin promoter activity in its normal cellular and genomic context. This cell-based system will be tested with compounds known to increase 17 integrin gene expression. The validated cell-based assay will then be applied to an automated high throughput drug screen to identify compounds that increase 17 integrin gene expression in cultured mouse muscle cells. Positive compounds will be tested in cultured murine and human muscle cells to determine drug efficacy, toxicity and ability to translate to human muscle cells. Together these aims will allow us to test the hypothesis that small compounds can increase 17 integrin gene expression in murine and human muscle cells. The identification of small compounds that increase 17 integrin gene expression will form the basis of future studies which will translate our basic biomedical research studies to the development of drugs for future human clinical trails.Project Narrative Duchenne Muscular Dystrophy is a devastating muscle disease that affects nearly 50,000 children in the United States. Increased expression of 17 integrin has been shown to be enormously beneficial to mouse models for this disease. This study aims to identify drugs that increase 17 integrin gene expression which may prove to be of therapeutic value to patients that suffer from DMD.