Regulation of iNOS gene expression in human T cells

人类 T 细胞 iNOS 基因表达的调控

基本信息

批准号：
371596-2009
负责人：
Choy, Jonathan
金额：
$ 2.19万
依托单位：
Simon Fraser University
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2012
资助国家：
加拿大
起止时间：
2012-01-01 至 2013-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=498056
关键词：
Regulation iNOS gene expression human

项目摘要

Nitric oxide (NO) is a bioactive gas involved in many aspects of human physiology. It is synthesized in cells by three related enzymes: neuronal nitric oxide synthase, inducible nitric oxide synthase (iNOS), and endothelial NOS. While all of these enzymes are important in the generation of NO under different biological circumstances, I have recently identified a role for iNOS expression in human T cells in the augmentation of human immune responses and shown that the chemokine stromal cell derived factor-1alpha (SDF-1a) is a protein inducer of iNOS in human T cells. However, the intracellular mechanisms regulating iNOS expression in T cells are unknown. Also, the regulation of iNOS expression in human cells, in general, is incompletely defined and is very different than that in animals commonly used to study mammalian biology, namely mice and rats. Therefore, the goal of my proposal is to understand the regulation of SDF-1a-induced iNOS expression in human T cells, and the role this plays in human T cell biology. The first part will study the intracellular signaling pathways that regulate SDF-1a-induced iNOS expression in human T cells. This will involve the inhibition of expression of the signaling enzymes ERK1/2 and PI3K by siRNA molecules, and then examination of SDF-1a-induced iNOS expression in human T cells lacking expression of ERK1/2 and PI3K. The role of Ras in mediating a potential feedback regulatory loop will also be examined. The second part of the proposal will determine the DNA elements and related transcription factors controlling the expression of the iNOS gene in human T cells. Finally, the third part of the proposal will examine the role of SDF-1a-induced iNOS in human T cell survival and migration. I have recently found that NO can inhibit cell death of human T cells and I will examine whether NO produced from iNOS in response to SDF-1a acts in a similar manner. The described work will be of interest to cell biologists and immunologists as it will provide new information on the regulation of iNOS expression in human cells, and insight into T cell biology as it relates to human immunity.

一氧化氮（NO）是与人类生理许多方面有关的生物活性气体。它是通过三种相关酶在细胞中合成的：神经元一氧化氮合酶，可诱导的一氧化氮合酶（iNOS）和内皮NOS。虽然所有这些酶在不同的生物学情况下的无生成中都很重要，但我最近确定了人类T细胞中iNOS表达的作用在人类免疫反应增强中的作用，并表明趋化因子基质细胞衍生因子-1alpha（SDF-1A）是人类T细胞中inos的蛋白质诱导剂。但是，调节T细胞中iNOS表达的细胞内机制尚不清楚。同样，通常，人类细胞中iNOS表达的调节是未完全定义的，与通常用于研究哺乳动物生物学的动物，即小鼠和大鼠。因此，我的建议的目的是了解人类T细胞中SDF-1A诱导的iNOS表达的调节，以及这种在人T细胞生物学中所扮演的作用。第一部分将研究调节人类T细胞中SDF-1A诱导的INOS表达的细胞内信号传导途径。这将涉及抑制信号传导酶ERK1/2和siRNA分子PI3K的表达，然后在缺乏ERK1/2和PI3K表达的人T细胞中检查SDF-1A诱导的INOS表达。还将检查RAS在介导潜在反馈调节循环中的作用。该提案的第二部分将确定控制人T细胞中iNOS基因表达的DNA元素和相关转录因子。最后，该提案的第三部分将研究SDF-1A诱导的iNOS在人类T细胞存活和迁移中的作用。我最近发现，没有人可以抑制人T细胞的细胞死亡，我将检查INOS是否以类似方式对SDF-1A起作用的INOS是否产生。所描述的工作将引起细胞生物学家和免疫学家的关注，因为它将提供有关人类细胞中iNOS表达的调节的新信息，并洞悉与人类免疫相关的T细胞生物学。