Methodology Core

方法论核心

• 批准号: 7845529
• 负责人: Sheila Muldoon
• 金额: $ 12.04万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7845529
• 关键词: 19q12; 1q31; Affect; Agonist; American; Anesthesia procedures; Anesthesiology; Anesthetics; B-Lymphocytes; Biochemical; Biopsy; Blood; Blood Tests; Blood specimen; Caffeine; Calcium; Cell Line; Central Core Myopathy; Chromosomes; Clinical; Code; Collaborations; Complementary DNA; Contracture; Coupling; Creatine Kinase; DNA; DNA-Directed DNA Polymerase; Dantrolene; Data; Databases; Defect; Dendritic Cells; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Digestion; Dihydropyridine Receptors; Disease; Enzymes; Event; Exercise; Exercise Tolerance; Exons; Exposure to; Family member; Fever; Freezing; Genes; Genetic; Genetic Screening; Genomics; Genotype; Goals; Halothane; Health Sciences; Heating; High Pressure Liquid Chromatography; Hot Spot; Human; Individual; Information Systems; Investigation; Laboratories; Lead; Life; Malignant hyperpyrexia due to anesthesia; Measurement; Measures; Mediating; Messenger RNA; Methodology; Methods; Molecular; Molecular Genetics; Molecular Profiling; Morbidity - disease rate; Muscle; Mutate; Mutation; Myoblasts; Neuromuscular Depolarizing Agents; Normal Range; Operative Surgical Procedures; Patients; Performance; Pharmaceutical Preparations; Phenotype; Physiological; Polymerase Chain Reaction; Predisposition; Preparation; Principal Investigator; Property; RNA analysis; Reaction; Receptor Gene; Recording of previous events; Registries; Research Personnel; Research Project Grants; Rhabdomyolysis; Risk Factors; RyR1; Ryanodine; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Sampling; Sarcoplasmic Reticulum; Screening procedure; Serum; Services; Signal Transduction; Skeletal Muscle; Specificity; Specimen; Strenuous Exercise; Succinylcholine; Syndrome; Test Result; Testing; Time; Tissue Sample; Translations; Universities; Update; base; genetic analysis; genetic technology; human tissue; improved; in vitro Model; mRNA Expression; mortality; mouse model; proband; programs; research clinical testing; response

Malignant Hyperthermia (MH) is an autosomal dominant disorder of skeletal muscle that causes a life threatening reaction following administration of commonly used inhalational anesthetics such as halothane, or the depolarizing muscle relaxant succinylcholine. Administration of the anesthetics during surgery causes an altered release of calcium from the sarcoplasmic reticulum (SR) through the ryanodine receptor (RYR1), which in turn induces a cascade of biochemical events that culminates in a nypermetabolic state (fulminant episode) with hyperthermia. If not treated, promptly by withdrawing the anesthetic and administering dantrolene (a drug that inhibits release of Ca++ from the SR) mortality can be as great as 70%. Because most MH individuals appear normal and susceptibility becomes apparent only after exposure to anesthetics, identifying MH susceptible individuals prior to surgery is difficult requiring muscle biopsy and measurements of the contractile sensitivity to RYR1 agonists caffeine and halothane. The caffeine-halothane contracture test (CHCT) phenotypes patients as MHS (susceptible) or MHN (normal). The CHCT has a sensitivity of 93% and specificity of 78%. USUHS has performed over 400 CHCT tests on patients. Approximately 50% of patients are diagnosed CHCT positive. Core C will review MH susceptible individuals, probands and family members for clinical histories of adverse responses to anesthetic drugs, phenotype skeletal muscle samples with performance of a CHCT, and genotype CHCT positive patients for causative mutations in RyR1 and the a1s-DHPR genes. They will collect blood samples from these patients a) for mRNA for use in profiling studies, b) to make permanent B-lymphocyte cell lines for physiologic and transcriptional profiling studies, and c) for the isolation of dendritic cells. In addition, they will collect muscle samples to make myoblast cultures to be purified by Core B, and flash freeze discarded muscle samples for transcriptional profiling studies.

恶性高热是一种常染色体显性遗传的骨骼肌疾病 在施用常用的药物后引起危及生命的反应， 吸入麻醉剂如氟烷，或去极化肌肉松弛剂 琥珀胆碱在手术过程中给予麻醉剂会改变 通过兰尼碱受体从肌浆网（SR）释放钙 （RYR 1），这反过来又诱导了一系列生化事件，最终导致 高代谢状态（暴发性发作）伴高热。如果不及时治疗， 撤回麻醉剂和管理丹曲林（药物，抑制释放的 来自SR的Ca++）死亡率可高达70%。因为大多数MH个体 只有在暴露于麻醉剂后才表现正常和敏感性变得明显， 在手术前识别MH易感个体是困难的，需要肌肉活检 以及测量对RYR 1激动剂咖啡因和氟烷的收缩敏感性。 咖啡因-氟烷挛缩试验（CHCT）将患者表型定为MHS （敏感）或MHN（正常）。CHCT的敏感性为93%，特异性为78%。 USUHS已经对患者进行了400多次CHCT测试。大约50%的患者 被诊断为CHCT阳性核心C将审查MH易感个体、先证者 和家庭成员对麻醉药物不良反应的临床史， 具有CHCT表现的表型骨骼肌样本和CHCT基因型 RyR 1和a1 s-DHPR基因致病突变阳性患者。他们将 从这些患者收集血液样品a）用于mRNA以用于谱分析研究，B）以 制备用于生理和转录谱分析的永久性B淋巴细胞系 研究，和c）用于树突细胞的分离。此外，他们将收集肌肉 通过Core B纯化用于制备成肌细胞培养物的样品，并弃去快速冷冻 用于转录谱研究的肌肉样本。