GENE EXPRESSION IN MATURE NEUTROPHILS

成熟中性粒细胞的基因表达

• 批准号: 2843565
• 负责人: PETER E NEWBURGER
• 金额: $ 46.07万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2843565
• 关键词: antiinflammatory agents; chemical stability; gene expression; genetic library; genetic regulation; genetic transcription; human subject; inflammation; leukocyte activation /transformation; messenger RNA; neutrophil; phlebotomy; restriction mapping; tissue /cell culture; transfection

DESCRIPTION (Adapted from Applicant's Abstract): Neutrophils provide an essential defense against infection, and play a major role in the tissue damage caused by inflammation. They also provide an excellent, simplified model for studying the modulation of mRNA levels in normal mammalian cells. With the emergence of public databases containing very large numbers of genes and expressed sequence tags, research in human genetics is now turning from investigation of single genes to the global analysis of gene expression. The principal investigators propose to apply this important new approach to human neutrophils. Using methods based on display of cDNA 3' end fragments, generated by specific restriction enzymes, the investigators will semi-quantitatively assess the levels of most mRNAs found in activated neutrophils. The proposed studies will test the central hypothesis that neutrophils show complex, stimulus-specific patterns of genetic regulatory responses, which can be explained, at least in part, by combinatorial use of "subroutines" of coordinately regulated responses. The specific aims are: 1) Determine the pattern of gene expression by neutrophils stimulated by a wide variety of well- defined agonists, as well as identify new genes involved in the neutrophil phagocytic response; 2) Compare the patterns of response to simple agonists versus complex physiological activators, including bacterial and sterile inflammatory stimuli, both in vitro and in vivo; 3) Dissect the components of the responses to bacterial and sterile inflammatory stimuli by inhibition of specific receptors and signal transduction pathways, exposure to clinically important anti-inflammatory drugs, and determination of the temporal and functional order of the complex response; 4) Determine the molecular mechanisms underlying the changes in gene transcript levels, including both transcriptional control mechanisms and regulation of mRNA stability. These studies may provide insight into the genetic repertoire of known and new genes that contribute to the neutrophil response and hence provide targets for intervention in augmentation of host defense or amelioration of inflammation. Insight into transcriptional and post-transcriptional mechanisms of gene expression in this system may also be applicable to other, more complex, cell types and tissues.

描述（根据申请人的摘要改编）：中性粒细胞为感染提供了必不可少的防御，并在炎症引起的组织损伤中起着重要作用。它们还提供了一个出色的简化模型，用于研究正常哺乳动物细胞中mRNA水平的调节。随着包含大量基因和表达序列标签的公共数据库的出现，人类遗传学的研究现在正在从单个基因的研究转变为基因表达的全局分析。主要研究人员建议将这种重要的新方法应用于人类中性粒细胞。使用基于特定限制酶产生的cDNA 3'末端片段的方法，研究人员将半定量评估活化中性粒细胞中发现的大多数mRNA的水平。拟议的研究将检验中心假设，即中性粒细胞显示出遗传调节反应的复杂，刺激特异性的模式，这可以通过结合使用协同调节的反应的“亚例程”来解释。具体目的是：1）确定由多种定义良好的激动剂刺激的中性粒细胞确定基因表达的模式，并确定与中性粒细胞吞噬吞噬反应有关的新基因； 2）在体外和体内比较对简单激动剂与复杂生理激活剂的反应模式，包括细菌和无菌炎症刺激； 3）通过抑制特定受体和信号转导途径，暴露于临床上重要的抗炎药以及确定复杂反应的时间和功能顺序； 4）确定基因转录水平变化的分子机制，包括转录控制机制和mRNA稳定性的调节。这些研究可能会洞悉已知基因和新基因的遗传库，这些基因有助于中性粒细胞反应，因此提供了干预宿主防御或改善炎症的靶标。对该系统中基因表达的转录和转录后的转录和转录后的见解也可能适用于其他更复杂的细胞类型和组织。