Functional Genomics Of Critical Illness

前言和目的：微阵列技术和蛋白质组学方法有望成为理解复杂疾病功能基因组学的关键工具。危重或受伤的患者经常死于不完全了解的情况，如感染性休克、急性呼吸窘迫综合征，并最终死于多器官功能障碍综合征。宿主炎症途径被认为在这些综合征中起主要致病作用。在基础水平上，宿主反应的临床和生物学表现是由基因表达的数量和质量变化决定的，或者至少是反映出来的。因此，器官损伤综合征可能由其相关的基因表达改变模式来定义。 寡核苷酸微阵列可以同时测量数千个基因的mRNA水平的相对变化，提供转录组的全球快照。这一系列研究的目标包括：1)将病理生理学和/或干预措施与反应基因联系起来；2)确定对危重疾病重要的系统特征、信号通路和调控网络；3)发现生物标记物、预后指标和药物开发的潜在靶点。 进度： 开发处理各种样本类型的实验室程序，包括中性粒细胞、外周血单核细胞、T淋巴细胞、全血支气管肺泡灌洗、脾、肝、肺和心脏。 放大程序的初步测试。 对包括人类、小鼠和大鼠在内的几个物种使用寡核苷酸微阵列的经验。 建造和广泛测试数据分析管道。 建立数据库和基于网络的分析工具。 干扰素g与地塞米松在原代支气管上皮细胞中相互作用的研究。 鉴定与镰状细胞性贫血、正常志愿者的内毒素攻击和大鼠模型中日益严重的感染相关的表达谱。 建议的工作进程： 定义原代人类细胞，包括中性粒细胞、单核细胞和内皮细胞对内毒素和其他免疫激活剂的体外反应。 确定感染不同类型的细菌是否会在肺炎大鼠模型中产生生物体特异性特征。 利用具有特定基因缺陷的“基因敲除”小鼠，连续检测脓毒症引起的心肌功能障碍的严重程度、相关基因表达的变化和结果。 研究全血对天花疫苗的转录反应。 确定镰状细胞性贫血的疾病严重程度和预后的生物标志物。 检查宿主对严重急性呼吸系统综合症(SARS)病毒的反应，并将其与引起“普通感冒”的冠状病毒和其他呼吸道病原体进行比较。

Introduction and Objective: Microarray technology and proteomic approaches promise to become a pivotal tool in understanding the functional genomics of complex diseases. Critically ill or injured patients frequently die of incompletely understood conditions such as septic shock, acute respiratory distress syndrome, and ultimately multiple organ dysfunction syndrome. Host inflammatory pathways are thought to play major pathogenic roles in these syndromes. At a basic level, the clinical and biological manifestations of host responses are determined or at least reflected by quantitative and qualitative changes in gene expression. Therefore, organ injury syndromes might be defined by their associated patterns of altered gene expression. Oligonucleotide microarrays can measure relative changes in mRNA levels for thousands of genes simultaneously providing a global snapshot of the transcriptome. The goals of this line of investigation include: 1) to link pathophysiology and/or interventions with responding genes; 2) identify system characteristics, signaling pathways, and regulatory networks important to critical illness; 3) discover biomarkers, prognostic indicators, and potential targets for drug development. Progress: Development of laboratory procedures for handling a variety of sample types including neutrophils, peripheral blood mononuclear cells, T-lymphocytes, whole blood broncheoalveolar lavage, spleen, liver, lung, and heart. Preliminary testing of amplication procedures. Experience with oligonucleotide microarrays for several species including human, mouse and rat. Construction and extensive testing of a data analysis pipeline. Establishment of a database and web based analytical tools. Investigation of interactions between interferon g and dexamethasone in primary bronchial epithelial cells. Identification of the expression profiles associated with sickle cell anemia, endotoxin challenge in normal volunteers, and escalating severity of infection in a rat model. Proposed Course of Work: Define the in vitro response of primary human cells including neutrophils, monocytes, and endothelial cells to endotoxin and other immune activators. Determine whether infection with different types of bacteria produce organism-specific signatures in a rat model of pneumonia. Using "knockout" mice with specific genetic defects serially examine the severity of sepsis-induced myocardial dysfunction, associated changes in gene expression, and outcome. Investigate the transcriptomic response of whole blood to smallpox vaccination. Identify biomarkers of disease severity and outcome in sickle cell anemia. Examine the host response to Severe Acute Respiratory Syndrome (SARS) virus in comparison to "common cold"-causing coronaviruses and other respiratory pathogens.