Molecular Characterization of Extraocular Muscle (EOM)

描述（由申请人提供）：长期目标是在分子水平上全面表征眼外肌（EOMs）。EOMs的精确功能是人类最佳视觉的绝对要求。EOMs是高度专门化的，具有广泛的反射/运动，其详细特性与其他肌肉不同。令人费解的是，eom对某些疾病有不同的敏感性。eom在重症肌无力、Grave's病和线粒体肌病中有显著的参与；然而，它们在杜氏肌营养不良症（DMD）中幸免，尽管所有其他骨骼肌群都广泛参与。我们的中心论点是，骨骼肌与其他骨骼肌有着根本的不同；我们可以将它们独特的特性和疾病易感性追溯到它们分子成分的独特模式；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独特分子特征的全局图像。虽然支持我们的中心论点，但分子表征尚未完成。关于新发现的调控分子（如microRNAs）在EOMs中的差异表达，亚细胞水平的表达差异（如亚突触或NMJ区域表达）或EOMs与肢体肌肉中差异基因表达的功能后果（如改变钙处理）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“miRNAome”，验证miRNA表达差异并创建EOM mRNA-miRNA相互作用组数据库；b)确定EOM亚突触（NMJ）区域的转录组，并确定由于视觉剥夺而导致的亚突触转录组的变化，c)表征EOM中钙调节分子的表达和钙稳态机制。这个项目与健康的关系是，通过完成这些目标，我们将确保更清楚地了解EOMs的分子和功能多样性，它们的生物力学特性，洞察这些独特肌肉的病理生理学和疾病的治疗发展。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EOMs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.