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

描述（由申请人提供）：杜氏肌营养不良症（DMD）是一种严重的肌肉萎缩性疾病，可导致过早死亡。目前，没有治愈或有效治疗这种毁灭性的神经肌肉疾病。17整合素的表达在DMD患者和mdx小鼠模型的肌肉中增加。缺乏肌营养不良蛋白和17整合素的小鼠表现出严重的肌营养不良，并在4周龄前死亡。最近，我们证明了骨骼肌中整合素17的转基因表达增强可以增加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剂，并表明上调17整合素基因表达的化合物可作为DMD的潜在疗法。在这项研究中，我们将启动一个小分子发现计划，以确定目标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细胞和基因组背景下评估17整合素启动子活性。这种基于细胞的系统将用已知增加17整合素基因表达的化合物进行测试。然后将验证的基于细胞的测定应用于自动化高通量药物筛选，以鉴定在培养的小鼠肌肉细胞中增加17种整合素基因表达的化合物。将在培养的鼠和人肌细胞中测试阳性化合物，以确定药物功效、毒性和转化为人肌细胞的能力。总之，这些目标将使我们能够测试的假设，即小化合物可以增加17整合素基因在小鼠和人类肌肉细胞的表达。对增加17种整合素基因表达的小分子化合物的鉴定将成为未来研究的基础，这将使我们的基础生物医学研究转化为未来人类临床试验药物的开发。 杜氏肌营养不良症是一种毁灭性的肌肉疾病，影响了美国近5万名儿童。17整合素的表达增加已被证明对这种疾病的小鼠模型非常有益。本研究的目的是确定增加17整合素基因表达的药物，这可能被证明对DMD患者具有治疗价值。

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.