Validation of a Muscle-Specific Tissue Image Analysis Tool for Quantitative Assessment of Dystrophin Staining in Frozen Muscle Biopsies
Validation of a Muscle-Specific Tissue Image Analysis Tool for Quantitative Assessment of Dystrophin Staining in Frozen Muscle Biopsies
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DOI：
10.5858/arpa.2017-0536-oa
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发表时间：
2019-02-01
影响因子：
4.6
通讯作者：
Milici, Anthony, I
中科院分区：
文献类型：
--
作者：
Aeffner, Famke;Faelan, Crystal;Milici, Anthony, I
上下文。杜氏肌营养不良症是一种罕见的、进行性的、致命的神经肌肉疾病，由肌营养不良蛋白丢失引起。常见的研究性治疗方法旨在增加病变肌肉中肌营养不良蛋白的表达。一些临床试验包括评估新的肌营养不良蛋白的产生，作为疗效的替代生物标志物，它可以预测治疗的临床益处。建立免疫荧光扫描和数字图像分析工作流程，为肌肉冷冻切片中生物标志物阳性纤维百分比的染色强度评估提供客观方法。-通过严格合格的荧光扫描过程实现了最佳和可重复的数字图像捕获。扫描确认后，通过将训练有素的病理学家获得的高倍显微镜视野总数和肌营养不良蛋白阳性纤维计数与MuscleMap获得的数据进行比较，对MuscleMap （Flagship Biosciences, Westminster, Colorado）算法进行验证。接下来，该算法在Duchenne肌营养不良症、Becker肌营养不良症和对照患者免疫荧光标记的肌肉切片的整片图像上进行测试。-当在训练有素的病理学家的指导下使用时，数字图像分析工具符合预定义的验证标准，并证明与手动评估在功能和统计上等效。据我们所知，这项工作是第一次分别对免疫荧光扫描和数字组织图像分析工作流程进行鉴定和验证，以支持临床试验环境所需的严格性。-MuscleMap可以分析整个肌肉活检切片中的所有纤维，并提供逐纤维的数据。这将使未来的临床试验能够在更高的一致性和细节水平上客观地研究肌纤维的肌营养不良蛋白表达。
Context.-Duchenne muscular dystrophy is a rare, progressive, and fatal neuromuscular disease caused by dystrophin protein loss. Common investigational treatment approaches aim at increasing dystrophin expression in diseased muscle. Some clinical trials include assessments of novel dystrophin production as a surrogate biomarker of efficacy, which may predict a clinical benefit from treatment.Objectives.-To establish an immunofluorescent scanning and digital image analysis workflow that provides an objective approach for staining intensity assessment of the tion of the percentage of biomarker-positive fibers in muscle cryosections.Design.-Optimal and repeatable digital image capture was achieved by a rigorously qualified fluorescent scanning process. After scanning qualification, the MuscleMap (Flagship Biosciences, Westminster, Colorado) algorithm was validated by comparing high-power microscopic field total and dystrophin-positive fiber counts obtained by trained pathologists to data derived by MuscleMap. Next, the algorithm was tested on whole-slide images of immunofluorescent-labeled muscle sections from Duchenne muscular dystrophy, Becker muscular dystrophy, and control patients.Results.-When used under the guidance of a trained pathologist, the digital image analysis tool met predefined validation criteria and demonstrated functional and statistical equivalence with manual assessment. This work is the first, to our knowledge, to qualify and validate immunofluorescent scanning and digital tissue imageanalysis workflow, respectively, with the rigor required to support the clinical trial environments.Conclusions.-MuscleMap enables analysis of all fibers within an entire muscle biopsy section and provides data on a fiber-by-fiber basis. This will allow future clinical trials to objectively investigate myofibers' dystrophin expression at a greater level of consistency and detail.