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

基本信息

批准号：
10354464
负责人：
Marino Di Franco
金额：
$ 20.59万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10354464
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&apos 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 Longitudinal Studies Malignant hyperpyrexia due to anesthesia Measurement Measures Membrane Membrane Potentials Methods 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 base design design and construction disease phenotype 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

项目摘要

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.

抽象的肌病是由于编码调节其前RNA的离子通道或蛋白质的基因突变而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导致肌肉电生理学和兴奋 - 合同耦合。这些遗传疾病目前尚无治愈，严重影响患者的生活质量并跨越罕见疾病的患病率（例如低血压和高血肿周期性瘫痪，1：100000）最常见的肌病（例如营养不良的肌肉症，1-5：10000）。虽然他们的遗传原因很容易可识别的，了解通道病和剪接病的机制，并设计他们的声音疗法策略，要求在全能中进行详细的电生理研究有意义的人类细胞环境。这些研究虽然可行，但由于缺乏实际缺乏，阻碍了这些研究高通量方法可用于人类肌肉。动物模型用于绕过这个缺点通常无法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的实用电生理方法来克服这些局限性与活检中解剖的人类肌肉纤维面容使用。我们将结合革命性的实验腔室，来自两种不同的电生理方法的技术，以执行定量，状态 ART，在近乎理想的条件下50-400μm长50-400μm的纤维段的电生理研究。与以前不同方法，这种新方法很容易实现，用户友好，并为具有统计学意义的研究。我们的方法将允许逐局电生理研究和筛查急性作用药物，使患者特定治疗方案的设计与当前趋势一致在当代精确医学中。那么，我们期望我们的方法将对人类产生变革性的影响肌肉生理和病理生理学。