TNF-alpha to TGF-beta Signal Transduction

TNF-α 至 TGF-β 信号转导

• 批准号: 7354604
• 负责人: Arnold Ralph Brody
• 金额: $ 25.86万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7354604
• 关键词: asbestos; biological signal transduction; cell line; cell population study; endothelin; enzyme activity; enzyme inhibitors; enzyme mechanism; fibrogenesis; gel mobility shift assay; gene expression; genetic transcription; genetically modified animals; interleukin 1; laboratory mouse; macrophage; mesenchyme; mitogen activated protein kinase; peptides; platelet derived growth factor; pulmonary fibrosis /granuloma; respiratory epithelium; small interfering RNA; transforming growth factors; tumor necrosis factor alpha

DESCRIPTION (provided by applicant): Interstitial pulmonary fibrosis (IPF) has numerous etiologies and afflicts millions of individuals worldwide. Yet, there are no effective preventive agents or therapeutic approaches. This situation is due in large part to a lack of understanding of the fundamental molecular mechanisms that mediate the fibrogenic process. In the work proposed here, we have focused on two peptide factors that have been implicated by numerous researchers as central to the development of IPF. These factors are tumor necrosis factor alpha (TNF-a) and transforming growth factor beta one (TGF-B1). Our previous work and preliminary data show that TNF-a up-regulates the expression of TGF-B1 both in vitro and in vivo, but the basic mechanisms through which TNF-a influences TGF-B1 expression remain undefined. TNF-a has been considered by many to be a "master cytokine" that controls the expression of a number of biologically relevant molecules. In as much as TGF-B1 is considered to be a primary "fibrogenic factor", we have set forth the central hypothesis that TNF-a induces TGF-B1 production by transcriptional regulation through the MEK/ERK pathway in lung cells: and the pathway of fibrogenesis mediated by TNF-a through TGF-B1 can be interrupted by specific siRNAs that mediate the degradation of transcripts that code for TNF-a, MP-1- a key component of MEK/ERK phosphorylation, and TGF-B1. Specific Aim 1 will establish MEK/ERK transduction pathways from TNF-a to TGF-B1 and the transcriptional mechanisms of expression. Aim 2 will focus on other factors that could up-regulate TGF-B1 through this same pathway; and in Aim 3, we will learn if the epithelial and mesenchymal cells in vivo in the lung exhibit the same transduction and transcriptional pathways in health and disease. These studies will provide specific targets for potential therapeutic approaches through the use of small interfering RNAs (siRNA). Thus, in Aim 4, we will show that an siRNA we have designed blocks endogenous TNF-a gene expression. We propose to use this construct as well as an siRNA that degrades MP1 (a key protein in MEK/ERK phosphorylation), and finally an siRNA that will block TGF-B1 expression, and ultimately the development of IPF in our established animal models.

描述（由申请人提供）：间质性肺纤维化（IPF）有多种病因，并折磨着全世界数百万人。然而，目前还没有有效的预防药物或治疗方法。这种情况在很大程度上是由于缺乏对介导纤维化过程的基本分子机制的理解。在这里提出的工作中，我们将重点放在两个肽因子上，这些因子被许多研究人员认为是IPF发展的核心。这些因子是肿瘤坏死因子α (TNF-a)和转化生长因子β 1 （TGF-B1）。我们之前的工作和初步数据表明，TNF-a在体外和体内均上调TGF-B1的表达，但TNF-a影响TGF-B1表达的基本机制尚不明确。TNF-a被许多人认为是控制许多生物学相关分子表达的“主细胞因子”。由于TGF-B1被认为是主要的“纤维化因子”，我们提出了中心假设，即TNF-a通过肺细胞中MEK/ERK通路的转录调节诱导TGF-B1的产生：并且TNF-a通过TGF-B1介导的纤维化途径可以被特定的sirna打断，这些sirna介导编码TNF-a、MP-1 （MEK/ERK磷酸化的关键成分）和TGF-B1的转录本的降解。Specific Aim 1将建立MEK/ERK从TNF-a到TGF-B1的转导途径及其表达的转录机制。Aim 2将关注通过相同途径上调TGF-B1的其他因素；在目标3中，我们将了解肺内上皮细胞和间充质细胞在健康和疾病中是否表现出相同的转导和转录途径。这些研究将通过使用小干扰rna （siRNA）为潜在的治疗方法提供特定的靶点。因此，在Aim 4中，我们将展示我们设计的siRNA阻断内源性TNF-a基因表达。我们建议使用这种结构以及降解MP1 （MEK/ERK磷酸化的关键蛋白）的siRNA，以及最终阻断TGF-B1表达的siRNA，并最终在我们已建立的动物模型中发展IPF。