Opto-Electrophysiological Method to Study Human Skeletal Muscle Channelopathies

研究人类骨骼肌通道病的光电生理学方法

• 批准号: 10596561
• 负责人: Marino Di Franco
• 金额: $ 15.44万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596561
• 关键词: Action Potentials; Acute; Affect; Animal Model; Animals; Behavior; Biochemical; Biopsy; Carbon Dioxide; Case Study; Cell membrane; Chlorides; Closure by clamp; Code; Coupling; Data; Dimensions; Disease; Drug Screening; Electrophysiology (science); Engineering; Ethics; Eulenburg' s Disease; Fiber; Frequencies; Functional disorder; Gene Expression; Genes; Genetic; Genetic Diseases; Goals; Human; Huntington Disease; Hyperkalemic periodic paralysis; Hypokalemic periodic paralysis; Impairment; Incisional Biopsy; Ion Channel; Kinetics; Laboratories; Lead; Life; Malignant hyperpyrexia due to anesthesia; Measurement; Measures; Membrane; Membrane Potentials; Methods; Modeling; Mus; Muscle; Muscle Fibers; Mutation; Myopathy; Myotonia Congenita; Myotonic Dystrophy; Noise; Optics; Organelles; Pathology; Patients; Pharmaceutical Preparations; Phenotype; Physiological; Potassium Channel; Prevalence; Proteins; Public Health; Pump; Quality of life; RNA; RNA Splicing; Rare Diseases; Risk; RyR1; Scheme; Signal Transduction; Skeletal Muscle; Sodium; Structure; Technology; Testing; Therapeutic; Variant; design; disease phenotype; drug action; human disease; human tissue; indium arsenide; individualized medicine; muscle physiology; mutant; novel; precision medicine; response; screening; sound; stem; trend; user-friendly; vastus lateralis; voltage clamp

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.

抽象的 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。表达、结构或功能异常 这些突变产生的离子通道会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 并涵盖罕见疾病的患病率范围（例如低钾性和高钾性周期性麻痹，1:100000） 最常见的肌病（例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潜在机制，并设计 对他们来说，合理的治疗策略需要在所有方面进行详细的电生理学研究 有意义的人类细胞背景。这些研究虽然可行，但由于普遍缺乏实用的、 适合用于人体肌肉的高通量方法。用于规避这一点的动物模型 缺陷往往无法重现大多数疾病或重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、实用的电生理学方法来克服这些限制 可以方便地用于从活组织检查中解剖的人体肌肉纤维。我们将结合革命性的实验 室采用两种不同的电生理方法技术来执行定量、状态 艺术，在近乎理想的条件下对 50-400μm 长的纤维段进行电生理学研究。与之前不同 方法，这种新方法易于实施、用户友好，并且提供了必要的高通量 具有统计学意义的研究。我们的方法将允许逐案进行电生理学研究和筛选 作用剧烈的药物，能够设计针对患者的特定治疗方案，符合当前趋势 在当代精准医学中。那么，我们期望我们的方法将对人类产生变革性影响 肌肉生理学和病理生理学。