Duchenne muscular dystrophy (DMD) is an incurable disease caused by out-of-frame DMD gene deletions while in frame deletions lead to the milder Becker muscular dystrophy (BMD). In the last decade several antisense oligonucleotides drugs have been developed to induce a partially functional internally deleted dystrophin, similar to that produced in BMD, and expected to ameliorate the disease course. The pattern of dystrophin expression and functionality in dystrophinopathy patients is variable due to multiple factors, such as molecular functionality of the dystrophin and its distribution. To benchmark the success of therapeutic intervention, a clear understanding of dystrophin expression patterns in dystrophinopathy patients is vital. Recently, several groups have used innovative techniques to quantify dystrophin in muscle biopsies of children but not in patients with milder BMD. This study reports on dystrophin expression using both Western blotting and an automated, high-throughput, image analysis platform in DMD, BMD, and intermediate DMD/BMD skeletal muscle biopsies. Our results found a significant correlation between Western blot and immunofluorescent quantification indicating consistency between the different methodologies. However, we identified significant inter- and intradisease heterogeneity of patterns of dystrophin expression in patients irrespective of the amount detected on blot, due to variability in both fluorescence intensity and dystrophin sarcolemmal circumference coverage. Our data highlight the heterogeneity of the pattern of dystrophin expression in BMD, which will assist the assessment of dystrophin restoration therapies.

中文翻译：

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高通量数字图像分析揭示了肌营养不良患者中肌营养不良蛋白表达的不同模式

杜氏肌营养不良症 (DMD) 是一种由框外 DMD 基因缺失引起的无法治愈的疾病，而框内缺失导致较轻微的贝克尔肌营养不良症 (BMD)。在过去的十年中，已经开发出几种反义寡核苷酸药物来诱导部分功能性的内部缺失的肌营养不良蛋白，类似于 BMD 中产生的肌营养不良蛋白，并有望改善疾病进程。由于多种因素，例如肌营养不良蛋白的分子功能及其分布，肌营养不良蛋白病患者中肌营养不良蛋白的表达和功能模式是可变的。为了衡量治疗干预的成功，清楚了解肌营养不良蛋白病患者的肌营养不良蛋白表达模式至关重要。最近，几个小组已经使用创新技术来量化儿童肌肉活检中的肌营养不良蛋白，但在 BMD 较轻的患者中却没有。本研究报告了在 DMD、BMD 和中间 DMD/BMD 骨骼肌活检中使用蛋白质印迹法和自动化、高通量、图像分析平台来表达抗肌萎缩蛋白。我们的结果发现蛋白质印迹和免疫荧光定量之间存在显着相关性，表明不同方法之间的一致性。然而，由于荧光强度和肌营养不良蛋白肌膜周长覆盖率的变化，我们发现患者中肌营养不良蛋白表达模式的显着疾病间和疾病内异质性，无论在印迹上检测到的量如何。我们的数据突出了 BMD 中肌营养不良蛋白表达模式的异质性，