Signaling pathway of TNF-alpha production and Clostridium difficile infection (CD

TNF-α 产生和艰难梭菌感染 (CD) 的信号通路

基本信息

批准号：
8637068
负责人：
Xingmin Sun
金额：
$ 9.39万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8637068
关键词：
Accounting Antibiotics Antibodies Apoptosis Bacterial Infections CDC42 gene Cells Clostridium difficile Cytoskeleton Dendritic Cells Diarrhea Disease Enterocolitis Epithelial Epithelial Cells Europe Event Exotoxins Glucans Goals Hospitalization Immune Immune response Immunohistochemistry In Situ In Vitro Incidence Infection Inflammation Inflammation Mediators Inflammatory Bowel Diseases Inflammatory Response Inflammatory disease of the intestine Intervention Intestines Knockout Mice Knowledge Lead MEKs Measures Mediating Mitogen-Activated Protein Kinases Modeling Molecular Morbidity - disease rate Mucositis Mus National Institute of Diabetes and Digestive and Kidney Diseases North America Pathway interactions Peritoneal Macrophages Phosphoric Monoester Hydrolases Play Prevention therapy Production Proteins Pseudomembranous Colitis Recurrence Resistance Reverse Transcriptase Polymerase Chain Reaction Role Severities Signal Pathway Signal Transduction Small Interfering RNA Specificity Staining method Stains System TNF gene Testing Tight Junctions Toxin Tumor Necrosis Factor Receptor Tumor Necrosis Factor-alpha Western Blotting Wild Type Mouse Work base career cell injury cytokine design enteric pathogen human MAPK14 protein human TNF protein in vivo macrophage mortality mouse model new therapeutic target novel particle response rho GTP-Binding Proteins

项目摘要

DESCRIPTION (provided by applicant): This is a K01 application from NIDDK in the field of Clostridium difficile infection (CDI). My long- term career objective is to utilize knowledge acquired through my work and others to develop measures of prevention and therapy against CDI and other enteric pathogens-associated diseases. The object of this study is to elucidate the signaling pathway of C. difficile toxin- mediated TNF-¿ production and develop a novel therapy against CDI by targeted blocking TNF- ¿ production. Three specific aims will be pursued. Specific Aim 1: Identify subsets of immune cells that are major producer of TNF-¿ in response to C. difficile toxins in vivo, based on our hypothesis that intestinal dendritic cells (DCs) and macrophages are major TNF-¿ producer during the bacterial infection. Specific Aim 2: Investigate the signaling events that lead to toxin-mediated TNF-¿ production in macrophages. Specific Aim 3: Evaluate blocking TNF-¿ production as an adjunctive therapy against intestinal inflammation in both primary and recurrent CDI. For specific aim1, TNF-¿ producing immune cells will be identified by immuofluorescence staining and immunohistochemistry in mouse ileal loop model and C. difficile infection in mice. To more precisely assess the roles of DCs and macrophages in TNF-¿ production, DC- or macrophage- depleted mice will be used. For specific aim 2, multiple approaches including siRNA knockdown, Western-blot analysis, RT-PCR will be performed to identify the involvement of small Rho GTPases and dual specificity phosphatases (DUSPs) in C. difficile toxin-induced TNF-¿ production. In specific aim 3, a novel glucan particle (GP)- dependent siRNA delivery system specifically targeting macrophages and DCs will be employed to evaluate blocking TNF-¿ production as an adjunctive therapy against intestinal inflammation in CDI.

描述（由申请人提供）：这是 NIDDK 在艰难梭菌感染 (CDI) 领域的 K01 申请我的长期职业目标是利用通过我和其他人的工作获得的知识来制定针对 CDI 的预防和治疗措施。和其他肠道病原体相关疾病的目的是阐明艰难梭菌毒素介导的 TNF-¿通过靶向阻断 TNF-¿ 生产和开发针对 CDI 的新型疗法将追求三个具体目标具体目标 1：确定 TNF-¿ 的主要产生者的免疫细胞亚群。基于我们的假设，肠道树突状细胞 (DC) 和巨噬细胞是主要的 TNF-¿具体目标 2：研究导致毒素介导的 TNF-¿ 的信号事件。具体目标 3：评估阻断 TNF-¿生产作为针对原发性和复发性 CDI 肠道炎症的辅助疗法1，TNF-¿将通过小鼠回肠环模型和小鼠艰难梭菌感染中的免疫荧光染色和免疫组织化学来鉴定产生免疫细胞，以更精确地评估 DC 和巨噬细胞在 TNF-¿对于具体目标 2，将使用 DC 或巨噬细胞耗尽的小鼠进行生产，将进行多种方法，包括 siRNA 敲低、蛋白质印迹分析、RT-PCR 来鉴定小 Rho GTP 酶和双特异性磷酸酶 (DUSP) 的参与。在艰难梭菌毒素诱导的 TNF-¿在具体目标 3 中，将采用专门针对巨噬细胞和 DC 的新型葡聚糖颗粒 (GP) 依赖性 siRNA 递送系统来评估阻断 TNF-¿生产作为对抗 CDI 肠道炎症的辅助疗法。