Methodology Core

方法论核心

• 批准号: 8054287
• 负责人: Sheila Muldoon
• 金额: $ 11.92万
• 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/8054287
• 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 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.

恶性热病是一种常染色体显性遗传性骨骼肌疾病 在服用常用药物后会引起危及生命的反应 吸入麻醉剂，如氟烷或去极化肌松剂 琥珀酰胆碱。在手术中使用麻醉药会引起 肌浆网(SR)通过兰尼定受体释放钙 (RyR1)，这反过来又引发一连串的生化事件，最终导致 过度代谢状态(暴发性发作)伴体温过高。如果不治疗，立即由 停用麻醉剂并使用丹曲林(一种抑制 来自SR的Ca++)死亡率可高达70%。因为大多数MH个体 看起来正常，只有在接触麻醉剂后，敏感性才会变得明显， 很难在手术前确定MH的易感个体，需要肌肉活检 并测量对RYR1激动剂咖啡因和氟烷的收缩敏感性。 咖啡因-氟烷收缩试验(CHCT)表型为MHS (易感)或MHN(正常)。CHCT的敏感性为93%，特异性为78%。 USUHS已经在患者身上进行了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易感个体和先证者 和家属了解麻醉药物不良反应的临床病史， 具有CHCT表现的表型骨骼肌样本和CHCT基因型 RyR1和A1S-DHPR基因致病突变阳性患者。他们会 采集这些患者的血液样本a)用于分析研究的mRNA，b) 建立永久的B淋巴细胞系以进行生理和转录分析 C)树突状细胞的分离。此外，它们还会收集肌肉 用于制作成肌细胞培养物的样品将通过Core B纯化，并快速冷冻丢弃 用于转录图谱研究的肌肉样本。