Heterozygous MH knock-in mice model Human MH susceptibility

杂合 MH 敲入小鼠模型 人类 MH 易感性

• 批准号: 7436116
• 负责人: Paul D Allen
• 金额: $ 21.91万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7436116
• 关键词: Affect; Bioinformatics; Biological Markers; Biopsy Specimen; Birth; Blood Screening; Blood specimen; Calcium; Cells; Central Core Myopathy; Clinical; Collaborations; Contracture; Coupled; Coupling; Dantrolene; Data; Diagnostic tests; Disease susceptibility; Functional disorder; Future; Gene Expression Profile; Genetic; Genotype; Goals; Halothane; Homeostasis; Hot Spot; Human; In Vitro; Invasive; Knock-in Mouse; Logistics; Malignant hyperpyrexia due to anesthesia; Marines; Membrane; Modeling; Molecular; Mus; Muscle; Muscle Fibers; Mutation; Myopathy; Pathology; Pathway interactions; Patients; Phenotype; Physiological; Predisposition; Principal Investigator; Process; Property; RNA; Receiver Operating Characteristics; Regression Analysis; Rest; Reverse Transcriptase Polymerase Chain Reaction; RyR1; Ryanodine Receptor Calcium Release Channel; Sampling; Screening procedure; Sensitivity and Specificity; Severities; Signal Transduction; Skeletal system; Specificity; Stress; Testing; Whole Blood; base; bench to bedside; congenic; design; human disease; in vivo; mouse model; peripheral blood; programs

The long-term goal of Project 1 is to define the mechanisms responsible for the ryanodine receptor (RyR1) myopathies, malignant hyperthermia and central core disease (MH/CCD). Our immediate objectives are to create suitable models for human disease in mice and use them to study how mutations of RyR1 alter intracellular Ca2+ homeostasis and to create a noninvasive diagnostic test for MH that has a high degree of specificity and sensitivity. HYPOTHESIS I: Heterozygous MH "knock-in" mice model Human MH susceptibility. A 1.1. To create a "hot-spot" region 2 MH 'knock in' mouse line (RyR1 G2434R). In addition to our recently created R163C (region 1) and T4826I (region 3) RyR1 MHS mice. A 1.2.To determine the relationship between the clinical MH phenotype and myoplasmic resting free calcium ([Ca2+],) in vivo, with contracture properties in vitro for each heterozygous MH/CCD mouse. A1.3 To determine the relationship between IVCT and myoplasmic resting [Ca2+] in myotubes from human biopsy samples. A1.4 To determine if genetic background can rescue the birth lethal phenotype seen in homozygous MH/CCD mice. A1.5 To determine if genetic background will lower or raise [Ca2+]i or sensitivity to halothane in heterozygous MH/CCD mice. HYPOTHESIS II: Mutations responsible for human MH/CCD increase passive RyR1 "leak" and alter the dynamics of EC coupling by enhancing ECCE and SOCE. A 2.1 To analyze heterozygous and homozygous MH/CCD myotubes for abnormalities in EC coupling and two forms of Ca2+ entry, ECCE and SOCE. A 2.2 To establish how halothane and dantrolene enhance and diminish aberrant Ca2+ signaling in MH/CCD myotubes. A 2.3 To validate murine MH/CCD myotube model results with myotubes obtained from humans with an analogous mutation. Hypothesis III: The "Funnel" approach can be used to create a non-invasive screening test for MH that has both a high degree of specificity and a high degree of sensitivity. A 3.1. We will analyze whole blood transcriptional profiles from patients with known MHS RyR1 mutations and use the Funnel approach to select a set of predictive markers to be used for future MHS screening.

项目1的长期目标是确定ryanodine受体（RyR1）的作用机制。