Molecular Characterization of Extraocular Muscle (EOM)

描述（由申请方提供）：长期目标是在分子水平上全面表征眼外肌（EOM）。EOM的精确功能是人类获得最佳视力的绝对要求。眼外肌是高度专业化的，并经历广泛的反射/运动，并且眼外肌的详细特性不同于其他肌肉的特性。令人费解的是，眼外肌对某些疾病具有不同的敏感性。眼外肌在重症肌无力、格雷夫斯病和线粒体肌病中有突出的参与;然而，它们在杜氏肌营养不良症（DMD）中幸免，尽管广泛参与所有其他骨骼肌群。我们的中心论点是，眼外肌是从根本上不同于其他骨骼肌;我们可以跟踪其独特的性质和疾病易感性的独特模式，其分子成分;一个“分子条形码”。在过去的资助期间，我们已经全面定义了EOM的“转录组”和“蛋白质组”。这些研究为我们提供了许多关于EOM功能的见解，并为我们提供了EOM独特分子特征的全球图景。虽然支持我们的中心论点，分子表征是尚未完成。关于新发现的调节分子（例如microRNA）类别的EOM中的差异表达、亚细胞水平的表达差异（例如突触下或NMJ区域表达）或EOM与肢体肌肉中差异基因表达的实际功能后果（例如改变的钙处理）的许多假设仍未得到解决。为了验证这些假设，我们提出：a）定义EOM“miRNA组”，验证miRNA表达差异并创建EOM mRNA-miRNA相互作用组数据库; B）定义EOM突触下（NMJ）区域的转录组并鉴定由于视觉剥夺引起的突触下转录组的改变，以及c）表征EOM中钙调节分子的表达和钙稳态机制。该项目的健康相关性在于，通过完成这些目标，我们将确保更清楚地了解EOM的分子和功能多样性，它们的生物力学特性，对这些独特肌肉的病理生理学和疾病治疗发展的见解。公共卫生相关性：该项目的健康相关性在于，通过完成我们的目标，我们将确保更清楚地了解EOM的分子和功能多样性以及它们的生物力学特性，并深入了解这些独特的肌肉。

DESCRIPTION (provided by applicant): The long-term goals are to comprehensively characterize extraocular muscles (EOMs) at the molecular level. Precise functioning of EOMs is an absolute requirement for optimal vision in humans. EOMs are highly specialized and undergo a wide range of reflexes/motions and the detailed properties of EOMs are different from those of other muscles. Enigmatically, EOMs have differential sensitivity to certain diseases. EOMs have prominent involvement in myasthenia gravis, Grave's disease and mitochondrial myopathies; they are spared, however, in Duchenne muscular dystrophy (DMD), despite the widespread involvement of all other skeletal muscle groups. Our central thesis is that EOMs are fundamentally different from other skeletal muscles; and that we can trace their unique properties and disease susceptibilities to unique patterns of their molecular constituents; a 'molecular barcode'. During the past funding period we have comprehensively defined the EOM 'transcriptome' and 'proteome'. These studies provide us with many insights into EOM function and provide us with a global picture of the unique molecular signature of EOM. While supporting our central thesis, the molecular characterization is as yet incomplete. A number of hypotheses remain unaddressed regarding differential expression in EOMs of newly discovered classes of regulatory molecules (e.g. microRNAs), expression differences at the subcellular level (e.g. sub-synaptic or NMJ regional expression) or indeed functional consequences of differential gene expression (e.g. altered calcium handling) in EOMs vs limb muscles. To test these hypotheses we propose: a) to define the EOM 'miRNAome', validate miRNA expression differences and create an EOM mRNA-miRNA interactome database; b) to define the transcriptome at the sub-synaptic (NMJ) regions of EOM and identify alterations to the sub-synaptic transcriptome due to visual deprivation, and, c) characterize expression of calcium regulatory molecules and calcium homeostatic mechanisms in EOM. The health relatedness of this project is that by completion of these aims we will ensure clearer understanding of the molecular and functional diversity of EOMs, their biomechanical properties, insights into pathophysiology of these unique muscles and therapeutic development for disease. PUBLIC HEALTH RELEVANCE: The health relatedness of this project is that by completion of our aims we will ensure clearer understanding of the molecular and functional diversity of EOMs as well as their biomechanical properties as well as insights into of these unique muscles.