Immunomodulatory mechanism of volatile anesthetics

挥发性麻醉药的免疫调节机制

基本信息

批准号：
8636035
负责人：
Koichi Yuki
金额：
$ 19.76万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8636035
关键词：
Absence of pain sensation Address Adhesions Anesthetics Animals Binding Binding Sites Biological Biological Assay Biological Process Biological Testing Boston Calorimetry Cell Adhesion Molecules Cells Clinical Critical Illness Data Disease Endothelium Fc Receptor Flow Cytometry Foundations Future General Anesthesia Goals Grant Health Care Costs Healthcare Hypnosis Immune Immune System Diseases Impairment Incidence Infection Infection Control Inflammation Institutes Integrins Intercellular adhesion molecule 1 Isoflurane K-Series Research Career Programs Knowledge Lead Length of Stay Leukocytes Ligand Binding Ligand Binding Domain Ligands Literature Measures Mediating Modification Molecular Molecular Medicine Natural Killer Cells Operative Surgical Procedures Outcome Patients Pediatric Hospitals Perioperative Phagocytosis Pharmaceutical Preparations Play Postoperative Complications Postoperative Period Proteins Public Health Research Resources Role Site Stereoisomer Structure Testing Titrations United States X-Ray Crystallography base cancer recurrence cell killing clinically significant cytotoxicity desflurane design extracellular high risk immune function immunological synapse formation immunoregulation insight mortality neutrophil programs research study sevoflurane

项目摘要

DESCRIPTION (provided by applicant): Volatile anesthetics are major drugs to provide general anesthesia during perioperative period. While the primary purpose is to provide analgesia, hypnosis and immobility, they are suggested to possess immunomodulatory effects. Alternation of immune function by volatile anesthetics can be clinically significant particularly fr critical-ill patients. Candidate's long-term goal is 1) to elucidate the molecular mechanism and degree of immunomodulatory effects by various, clinically-used anesthetics, thereby providing clinicians the rational to use one anesthetic over others in the context of patient's underlying diseases, and 2) to redesign new anesthetics without immunomodulation. We focused our target on two adhesion molecules L2 and M2 that are expressed exclusively on leukocytes, playing a role in leukocyte arrest on the endothelium, phagocytosis, immunological synapse formation, natural killer cells cytotoxicity, and so on. In our preliminary experiments, we showed that L2 and M2 are functionally inhibited by isoflurane. Also L2, not M2 was inhibited by sevoflurane. We hypothesize that volatile anesthetics bind to these integrins and modify their functions, thereby modulating immune function. In this proposal, we will investigate how volatile anesthetics interact with these molecules structurally. Our proposed research consists of three Aims, and will be performed in Immune Disease Institute (Program in Cellular and Molecular Medicine in Children's Hospital Boston). In Aim1 and Aim 2, we will determine the interaction of clinically used anesthetics with L2 and M2 and their binding site(s). The effects of volatile anesthetics on ligand binding will be assessed in cell-free and cell-based assays using flow cytometery and V-bottom assay. The binding parameters will be measured with isothermal titration calorimetry. Anesthetic binding site(s) will be assessed in the combination of X ray crystallography and photoactivatable anesthetics. In Aim3, we will examine biological effects by volatile anesthetics resulting from the impairment of these adhesion molecules' function. We will use natural killer cells cytotoxicity assay, Fc receptor mediated neutrophil cytotoxicity and phagocytosis assay for this purpose. Subsequently, our experimental results will provide insight into the underlying mechanism of volatile anesthetics-induced immunomodulation.

描述（由申请人提供）：挥发性麻醉是在围手术期间提供全身麻醉的主要药物。尽管主要目的是提供镇痛，催眠和固定性，但建议它们具有免疫调节作用。通过挥发性麻醉药的免疫功能的交替在临床上是显着的，尤其是关键的ILL患者。候选人的长期目标是1）阐明各种临床中使用的麻醉药的分子机制和免疫调节作用的分子机制和程度，从而为临床医生提供了在患者潜在疾病的背景下使用一种麻醉的临床医生，而2）在没有免疫原理的情况下重新设计了新的麻醉药。我们将目标集中在两个仅在白细胞上表达的粘附分子L2和M2，在内皮细胞上的白细胞停滞，吞噬作用，免疫学突触形成，天然杀伤细胞细胞细胞中发挥作用。在我们的初步实验中，我们表明L2和M2在功能上被异氟烷抑制。还有L2，而不是M2被Sevoflurane抑制。我们假设挥发性麻醉剂结合了这些整合素并修改其功能，从而调节免疫功能。在此提案中，我们将研究挥发性麻醉剂在结构上如何与这些分子相互作用。我们提出的研究包括三个目标，并将在免疫疾病研究所（波士顿儿童医院的细胞和分子医学计划）进行。在AIM1和AIM 2中，我们将确定临床使用麻醉药与L2和M2及其结合位点的相互作用。挥发性麻醉药对配体结合的影响将在无细胞和基于细胞的测定中使用流式细胞仪和V-Bottom测定法进行评估。结合参数将通过等温滴定量热法测量。麻醉结合位点将在X射线晶体学和光活化麻醉剂的组合中进行评估。在AIM3中，我们将通过这些粘附分子功能受损而导致的挥发性麻醉剂检查生物学作用。为此，我们将使用天然杀伤细胞的细胞毒性测定，FC受体介导的中性粒细胞毒性和吞噬作用分析。随后，我们的实验结果将提供有关挥发性麻醉剂引起的免疫调节的潜在机制的洞察力。