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）是一种生物活性气体，参与人体生理学的许多方面。它在细胞中由三种相关酶合成：神经元一氧化氮合酶、诱导型一氧化氮合酶 (iNOS) 和内皮一氧化氮合酶。虽然所有这些酶在不同的生物环境下对 NO 的生成都很重要，但我最近发现了人类 T 细胞中 iNOS 表达在增强人类免疫反应中的作用，并表明趋化因子基质细胞衍生因子 1α (SDF -1a) 是人类 T 细胞中 iNOS 的蛋白诱导剂。然而，调节 T 细胞中 iNOS 表达的细胞内机制尚不清楚。此外，一般来说，人类细胞中 iNOS 表达的调节尚未完全确定，并且与通常用于研究哺乳动物生物学的动物（即小鼠和大鼠）中的 iNOS 表达调节有很大不同。因此，我的建议的目标是了解人类 T 细胞中 SDF-1a 诱导的 iNOS 表达的调节，以及它在人类 T 细胞生物学中发挥的作用。第一部分将研究调节人类 T 细胞中 SDF-1a 诱导的 iNOS 表达的细胞内信号通路。这将涉及通过 siRNA 分子抑制信号酶 ERK1/2 和 PI3K 的表达，然后检查缺乏 ERK1/2 和 PI3K 表达的人类 T 细胞中 SDF-1a 诱导的 iNOS 表达。 Ras 在调节潜在反馈调节环路中的作用也将被研究。该提案的第二部分将确定控制人类 T 细胞中 iNOS 基因表达的 DNA 元件和相关转录因子。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NO可以抑制人类T细胞的细胞死亡，我将研究iNOS响应SDF-1a产生的NO是否以类似的方式发挥作用。所描述的工作将引起细胞生物学家和免疫学家的兴趣，因为它将提供有关人类细胞中 iNOS 表达调节的新信息，并深入了解与人类免疫相关的 T 细胞生物学。