Opto-Electrophysiological Method to Study Human Skeletal Muscle Channelopathies

摘要 由编码离子通道或调节其前RNA的蛋白质的基因突变引起的肌病 剪接分别称为通道病和剪接病。异常表达、结构或功能 这些突变导致的离子通道的变化导致肌肉电生理学和兴奋的改变- 收缩耦合这些遗传性疾病目前尚无治愈方法，严重影响患者的生活质量 从罕见病（如低钾和高钾性周期性麻痹，1：100000） 最常见的肌病（如营养不良性肌强直，1-5：10000）。虽然他们的遗传原因很容易 可识别，了解通道病和剪接病的机制，并设计 对他们的合理治疗策略，需要详细的电生理学研究，在所有的， 有意义的人类细胞背景。这些研究虽然可行，但由于普遍缺乏实际的、 适用于人体肌肉的高通量方法。动物模型用来规避这一点 这些缺点往往不能概括大多数疾病或再现人类对治疗药物的反应。 我们打算通过设计和测试一种新颖、实用的电生理方法来克服这些局限性 易于用于从活组织检查中解剖的人体肌肉纤维。我们将联合收割机 室技术从两个不同的电生理方法来执行定量，状态的 art，在接近理想的条件下对50-400μm长的纤维段进行电生理学研究。不同于以往 方法，这种新方法易于实施，用户友好，并提供了必要的高通量， 有统计意义的研究。我们的方法将允许逐个病例的电生理学研究和筛选 急性作用药物，使患者的具体治疗方案的设计，符合目前的趋势 在当代精准医疗中的应用我们希望，我们的方法将对人类产生变革性的影响， 肌肉生理学和病理生理学。

ABSTRACT Myopathies stemming from mutations in genes coding for ion channels or proteins that regulate their pre-RNA splicing are called, respectively, channelopathies and spliceopathies. Abnormal expression, structure or function of ion channels resulting from those mutations leads to altered muscle electrophysiology and excitation- contraction coupling. These genetic disorders have currently no cure, severely affect the quality of life of patients and span the prevalence spectrum from rare diseases (e.g. hypo- and hyperkalemic periodic paralysis, 1:100000) to the most common myopathies (e.g. dystrophic myotonia, 1-5:10000). While their genetic cause is readily identifiable, understanding the mechanisms underlying channelopathies and spliceopathies, and designing sound therapeutic strategies for them, demands detailed electrophysiological studies performed in the all- meaningful human cellular context. These studies, though feasible, are impeded by a pervasive lack of practical, high throughput methods amenable for use in human muscles. Animal models used to circumvent this shortcoming often fail to recapitulate most diseases or to reproduce the human response to therapeutic drugs. We intend to overcome these limitations by designing and testing a novel, practical electrophysiological method facilely used with human muscle fibers dissected from biopsies. We will combine a revolutionary experimental chamber with technologies from two different electrophysiological methods to perform quantitative, state of the art, electrophysiological studies in segments of fibers 50-400μm long in near-ideal conditions. Unlike previous methods, this new method is readily implemented, user-friendly, and affords the requisite high throughput for statistically significant studies. Our method will allow case-by-case electrophysiological studies and screening of acutely acting drugs, enabling the design of patient specific treatment schemes, coinciding with current trends in contemporary precision medicine. We expect, then, our method will have a transformative impact in human muscle physiology and pathophysiology.