权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

REGULATION OF CXC CHEMOKINE EXPRESSION BY H. PYLORI

幽门螺杆菌对 CXC 趋化因子表达的调节

基本信息

批准号：
8534093
负责人：
YOSHIO YAMAOKA
金额：
$ 31.58万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-06-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8534093
关键词：
Acute Animal Model Architecture Atrophic Beryllium Binding Sites Biological Assay Biology CXC Chemokines Cell Surface Receptors Cells Characteristics Chronic Clinical Complement Factor B Cytotoxin Data Development Disease Elements Epidermal Growth Factor Receptor Epithelial Epithelial Cells Epithelium Family G-Protein-Coupled Receptors Gastric Adenocarcinoma Gastric lymphoma Gastric mucosa Gastritis Genes Genetic Transcription Gerbils Goals Grant Helicobacter Infections Helicobacter pylori Human In Vitro Infection Infection prevention Infectious Agent Inflammation Inflammatory Injury Interferons Interleukin-6 Interleukin-8 Knock-in Mouse Laboratories Link Membrane Proteins Mitogen-Activated Protein Kinases Modeling Molecular Mus Nuclear PTPN11 gene Pathogenesis Pathogenicity Island Peptic Ulcer Phase Phosphoric Monoester Hydrolases Production Protein Family Proteins Reagent Regulation Role STAT protein Signal Pathway Signal Transduction Stomach Structure Techniques Therapeutic Tight Junctions Transcription Factor AP-1 Type IV Secretion System Pathway Ulcer Virulence Factors Virulent Work apical membrane base basolateral membrane carcinogenesis chemokine cytokine in vitro Model in vivo innovation insight malignant stomach neoplasm member migration mouse model neutrophil new therapeutic target novel promoter public health relevance receptor research study src Homology Domains tool transcription factor tumor

项目摘要

DESCRIPTION (provided by applicant): Helicobacter pylori (Hp) infection is the major cause of gastric inflammation, peptic ulcer disease, and gastric cancer. Recent clinical and in vitro data indicate that the Hp outer inflammatory protein, OipA and the cag pathogenicity island (PAI) are involved in disease pathogenesis based on their ability to induce interleukin (IL)-8 in gastric epithelial cells. Our long term goal is to understand the role(s) of OipA, alone and together with the cag PAI, in the pathogenesis of Hp-related diseases. In AIM 1, we will determine in vitro molecular mechanisms by which OipA, alone and together with the cag PAI, regulate IL-8 gene transcription. Binding sites for the transcription factors nuclear factor-:B (NF-:B), activator protein 1 (AP-1), interferon-stimulated responsive element (ISRE)-like element, and likely interferon-3 activation sequence (GAS) in the IL-8 promoter are involved in regulating IL-8 gene transcription in Hp-infected gastric epithelial cells. We will use polarized gastric epithelial cells with tight junctions (NCI-N87 cells) to overcome the limitations of non-polarized cells. NCI-N87 cells allow us to detect different cell signaling pathways from the apical membrane (early phase of infection) and basolateral membranes (late phase following opening of tight junctions or epithelial-barrier damage). We will focus on the cell surface receptors epidermal growth factor receptor (EGFR) and G protein coupled receptor (GPCR). In vitro models of Hp infection will be used to clarify 1) overall signaling pathways from the cell surface receptor(s) resulting in IL-8 gene transcription, 2) interactions between OipA and the cag PAI, and 3) OipA structures and motifs responsible for IL-8 gene transcription. In AIM 2, we will determine the correlation between in vitro - in vivo roles of OipA and the cag PAI in gastric injury using Mongolian gerbils and the gp130F759 knock-in mouse model. In gerbils, gastric mucosal transcription factors induced by Hp infection differ with respect to phase and effect on infection (e.g., AP-1 is induced early and results in inflammation and ulceration whereas NF- B and ISRE are induced late and result in atrophy). Our previous results are based on analysis of the whole gastric mucosa. Here, we propose to determine how OipA and the cag PAI induce CXC chemokines using isolated gastric epithelial cells. We will also use mice that lack the Src homology 2 domain of the Src homology 2 phosphatase (SHP-2) binding site on the IL-6 family co- receptor, gp130. In these mice, the signal transducer and activator of transcriptions (STATs) signaling is hyperactivated and SHP-2->AP-1 signaling is absent. Our preliminary data showing that Hp infection results in hyperproliferative tumors in this mouse model will be extended to further clarify the roles of OipA and the cag PAI in the gastroduodenal pathogenesis. We hypothesize that the early phase, where the epithelial cell-cell barrier is intact in vitro, will correlate with acute inflammation in vivo and the late phase, where the epithelial barrier is disrupted in vitro, will correlate with chronic inflammation, ulceration and carcinogenesis (i.e., when the architecture of the gastric epithelium is disrupted in vivo). The innovative focus and strategic approaches described in this application will yield new levels of understanding of the biology of Hp infections. PUBLIC HEALTH RELEVANCE: Helicobacter pylori infection causes gastric inflammation, peptic ulcer disease, and gastric cancer. The work outlined in this application will use human gastric cells and animal models to identify the molecular mechanisms by which Helicobacter pylori virulence factors cause gastric inflammation and subsequent disease. This project will be able to identify novel therapeutic targets for the treatment of Helicobacter pylori infections and prevention of Helicobacter pylori-related diseases.

描述（由申请人提供）：幽门螺杆菌（Hp）感染是胃炎症、消化性溃疡和胃癌的主要原因。最近的临床和体外研究表明，Hp外炎性蛋白、OipA和cag致病岛（派）通过诱导胃上皮细胞产生白细胞介素（IL）-8而参与疾病的发病机制。我们的长期目标是了解OipA单独或与cag派一起在Hp相关疾病发病机制中的作用。在AIM 1中，我们将确定OipA单独和与cag派一起调节IL-8基因转录的体外分子机制。IL-8启动子中转录因子核因子-：B（NF-：B）、激活蛋白1（AP-1）、干扰素刺激的应答元件（ISRE）样元件和可能的干扰素3激活序列（GAS）的结合位点参与调节Hp感染的胃上皮细胞中IL-8基因的转录。我们将使用具有紧密连接的极化胃上皮细胞（NCI-N87细胞）来克服非极化细胞的局限性。NCI-N87细胞使我们能够检测来自顶膜（感染的早期阶段）和基底外侧膜（紧密连接打开或上皮屏障损伤后的晚期阶段）的不同细胞信号传导途径。重点介绍了细胞表面受体表皮生长因子受体（EGFR）和G蛋白偶联受体（GPCR）。Hp感染的体外模型将用于阐明1）导致IL-8基因转录的细胞表面受体的总体信号传导途径，2）OipA和cag派之间的相互作用，以及3）负责IL-8基因转录的OipA结构和基序。在AIM 2中，我们将使用蒙古沙鼠和gp 130 F759基因敲入小鼠模型确定OipA和cag派在胃损伤中的体外-体内作用之间的相关性。在沙鼠中，由Hp感染诱导的胃粘膜转录因子在感染的阶段和影响方面不同（例如，AP-1被早期诱导并导致炎症和溃疡，而NF- B和ISRE被晚期诱导并导致萎缩）。我们以前的结果是基于对整个胃粘膜的分析。在这里，我们建议，以确定如何OipA和cag派诱导CXC趋化因子使用分离的胃上皮细胞。我们还将使用缺乏IL-6家族共受体gp 130上Src同源2磷酸酶（SHP-2）结合位点的Src同源2结构域的小鼠。在这些小鼠中，信号转导和转录激活因子（STAT）信号传导被过度激活，并且SHP-2->AP-1信号传导不存在。我们的初步数据表明，在这种小鼠模型中，Hp感染导致过度增殖性肿瘤，将进一步阐明OipA和cag派在胃十二指肠发病机制中的作用。我们假设早期阶段，其中上皮细胞-细胞屏障在体外是完整的，将与体内急性炎症相关，而晚期阶段，其中上皮屏障在体外被破坏，将与慢性炎症、溃疡和致癌相关（即，当胃上皮的结构在体内被破坏时）。本申请中描述的创新重点和战略方法将使人们对Hp感染生物学的理解达到新的水平。公共卫生相关性：幽门螺杆菌感染可引起胃炎症、消化性溃疡和胃癌。本申请中概述的工作将使用人类胃细胞和动物模型来鉴定幽门螺杆菌毒力因子引起胃炎症和随后疾病的分子机制。该项目将能够确定治疗幽门螺杆菌感染和预防幽门螺杆菌相关疾病的新的治疗靶点。