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：
8278863
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/8278863
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. PUBLIC HEALTH RELEVANCE: Immunomodulation by volatile anesthetics has been described in literature, but there is lack of knowledge how, which and when anesthetics will cause immunomodulation. We propose to address the mechanism of volatile anesthetics-induced immunomodulation. Understanding the mechanism, the degree, and significance of these effects is important to formulate our daily practice of general anesthesia, and potentially impact on the outcomes of patients undergoing surgery.

描述（申请人提供）：挥发性麻醉药是围手术期全身麻醉的主要药物。虽然主要目的是提供镇痛、催眠和固定，但它们被认为具有免疫调节作用。挥发性麻醉剂改变免疫功能可能具有临床意义，特别是对于危重患者。候选人的长期目标是 1) 阐明各种临床使用的麻醉剂的免疫调节作用的分子机制和程度，从而为临床医生提供在患者基础疾病的情况下使用一种麻醉剂而不是其他麻醉剂的合理性，以及 2) 重新设计无需免疫调节的新型麻醉剂。我们的目标集中在两种仅在白细胞上表达的粘附分子L2和M2，它们在内皮上的白细胞停滞、吞噬作用、免疫突触形成、自然杀伤细胞的细胞毒性等中发挥作用。在我们的初步实验中，我们表明 L2 和 M2 受到异氟烷的功能性抑制。 L2（而不是M2）也被七氟醚抑制。我们假设挥发性麻醉剂与这些整合素结合并改变它们的功能，从而调节免疫功能。在本提案中，我们将研究挥发性麻醉剂如何与这些分子在结构上相互作用。我们提出的研究包括三个目标，并将在免疫疾病研究所（波士顿儿童医院细胞和分子医学项目）进行。在目标 1 和目标 2 中，我们将确定临床使用的麻醉剂与 L2 和 M2 及其结合位点的相互作用。将使用流式细胞术和 V 底测定在无细胞和基于细胞的测定中评估挥发性麻醉剂对配体结合的影响。将用等温滴定量热法测量结合参数。麻醉剂结合位点将结合 X 射线晶体学和光活化麻醉剂进行评估。在 Aim3 中，我们将检查挥发性麻醉剂因这些粘附分子功能受损而产生的生物效应。为此，我们将使用自然杀伤细胞的细胞毒性测定、Fc 受体介导的中性粒细胞细胞毒性和吞噬作用测定。随后，我们的实验结果将深入了解挥发性麻醉剂诱导的免疫调节的潜在机制。公共卫生相关性：文献中已经描述了挥发性麻醉剂的免疫调节作用，但缺乏了解麻醉剂如何、哪种以及何时会引起免疫调节。我们建议解决挥发性麻醉药诱导的免疫调节的机制。了解这些影响的机制、程度和意义对于制定我们日常的全身麻醉实践非常重要，并且可能对接受手术的患者的结果产生影响。