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。这些突变产生的离子通道导致肌肉电生理学和兴奋- 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）很容易由遗传引起。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这些研究虽然可行，但由于普遍缺乏实用的、高通量方法适用于人体肌肉，用于规避这一点。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轻松地与从活组织检查中解剖的人体肌肉纤维一起使用，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。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，符合当前趋势那么，我们期望我们的方法将对当代精准医学产生变革性影响。肌肉生理学和病理生理学。