Regulation of IFN-beta induction by P2X7 purinergic receptor signaling

P2X7 嘌呤能受体信号传导对 IFN-β 诱导的调节

• 批准号: 7936194
• 负责人: Judith Anne Smith
• 金额: $ 13.99万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936194
• 关键词: Adenine Nucleotides; Agonist; Antiviral Agents; Area; Autoimmune Diseases; Autoimmune Process; Basic Science; Binding; Biology; Blood Platelets; CD14 Antigen; Calcium; Calcium Signaling; Cations; Cells; Cessation of life; Clinical; Collaborations; Complex; Continuing Education; Cytolysis; Cytoplasmic Tail; Data; Dendritic cell activation; Development Plans; Developmental Process; Disease; Double-Stranded RNA; Employee Strikes; Endotoxins; Environment; Equipment; Faculty; Family; Funding; G-Protein-Coupled Receptors; Genes; Genetic Transcription; Goals; Immune; Immune response; Infection; Inflammation; Inflammatory; Interferon-beta; Interferons; Lead; Ligands; Ligation; Lipopolysaccharides; MAP Kinase Activation Pathway; MAP Kinase Signaling Pathways; MAPK14 gene; MAPK8 gene; Mediating; Medical; Membrane; Mentors; Messenger RNA; Mitogen-Activated Protein Kinases; Modification; Morphogenesis; Mus; Natural Immunity; Nitric Oxide; Nucleotides; Osteitis; Osteoporosis; P2X-receptor; Pathway interactions; Physicians; Play; Principal Investigator; Process; Production; Purine Nucleotides; Purines; Purinoceptor; Receptor Signaling; Regulation; Research; Research Project Grants; Role; Schools; Scientist; Sepsis; Signal Induction; Signal Transduction; Signal Transduction Pathway; Site; Stimulus; Study Section; TLR4 gene; Techniques; Testing; Time; Tissues; Training; Universities; Virus Diseases; Wisconsin; Work; adaptive immunity; analog; bactericide; bone; career; career development; chromatin immunoprecipitation; cytokine; extracellular; immunoregulation; improved; interest; macrophage; member; novel; nucleotide analog; nucleotide receptor; pediatric department; programs; promoter; purine; receptor; release of sequestered calcium ion into cytoplasm; response; small molecule; transcription factor

DESCRIPTION (provided by applicant): Goals: My long-term research goal is to gain a greater understanding of the regulation of type I IFN production by macrophages, cells that are key players in inflammation. As a physician-scientist, it is my hope that these studies will eventually contribute to the improved treatment of inflammatory and autoimmune conditions. My immediate career goal is to develop a viable independent research program. Research project: During inflammation, the occurrence of tissue damage leads to sufficient release of extracellular purine nucleotides to stimulate immune cells via purinergic receptors such as P2X7. Stimulation of P2X7 by ATP is well known to increase the production of inflammatory cytokines (e.g. IL-1¿) by macrophages stimulated with endotoxin (LPS). We have found that co-stimulation of macrophages with LPS and the ATP analogue BzATP greatly augments the induction of IFN- ¿ by LPS. Type I IFNs play important roles in diverse aspects of innate and adaptive immunity. As both IFN- ¿ and purinergic receptor signaling have been implicated in bacterial sepsis and bone morphogenesis, it is critical to understand the mechanism of how P2X7 modulates the induction of IFN- ¿. To begin dissecting the mechanism of how P2X7 exerts its effect, a multi-pronged approach is proposed: We will examine the effect of P2X7 ligation on IFN- ¿ induction by different infectious stimuli (e.g. LPS, dsRNA etc.) to determine whether P2X7 enhancement is LPS specific. P2X7 ligation is known to initiate multiple signal transduction pathways: the contribution of calcium and MAP-kinase signaling pathways to the effect of BzATP on IFN- ¿ induction will be analyzed. Finally, we will examine the recruitment of transcription factors to the IFN- ¿ gene promoter by chromatin immunoprecipitation to test our underlying hypothesis that P2X7 ligation leads to increased transcription factor occupancy of the ifnb1 promoter. Modification of transcription factor recruitment by P2X7 mediated signaling has ramifications for the enhancement of immune responses and bone biology. Career development plan: The two key components are close collaboration with Paul Bertics and more formal oversight by a mentoring committee. Paul Bertics has abundant expertise in my area of interest and an excellent track record in training junior faculty. In addition to Dr. Bertics, my committee is composed of three other very supportive and successful physician scientists. Research environment: Through the department of Pediatrics, I have been accorded lab space, protected time and initial start-up funds. The breadth of expertise present at the University of Wisconsin, through the Medical and Graduate schools, is tremendous. Any equipment or technical assistance I should require will be available. There are ample forums for intellectual exchange as well as continuing education in the clinical and basic sciences. RELEVANCE: Macrophages participate in many inflammatory processes including responses to infections and autoimmune diseases by secreting cytokines. My research proposes to study how substances that are released by tissue damage called "purines" regulate the production of the inflammatory cytokine IFN- ¿ by macrophages. Greater understanding of inflammatory processes may ultimately lead to the better treatment of disease.

描述（由申请人提供）： 目标：我的长期研究目标是更好地了解巨噬细胞（炎症中的关键细胞）对 I 型干扰素产生的调节，作为一名医生科学家，这是我的希望。这些研究最终将有助于改善炎症和自身免疫性疾病的治疗，我的近期职业目标是开发一个可行的独立研究项目：在炎症过程中，组织损伤的发生导致细胞外嘌呤核苷酸的充分释放。众所周知，ATP 刺激 P2X7 会增加内毒素 (LPS) 刺激的巨噬细胞产生炎症细胞因子（例如 IL-1¿）。与 LPS 和 ATP 类似物 BzATP 一起大大增强 IFN-¿ I 型干扰素在先天性和适应性免疫的各个方面发挥着重要作用。由于嘌呤能受体信号传导与细菌性脓毒症和骨形态发生有关，因此了解 P2X7 如何调节 IFN-¿ 诱导的机制至关重要。为了开始剖析 P2X7 如何发挥其作用的机制，提出了一种多管齐下的方法：我们将检查 P2X7 连接对 IFN-¿通过不同的感染刺激（例如 LPS、dsRNA 等）进行诱导以确定 P2X7 增强是否是 LPS 特异性的，已知 P2X7 连接会启动多种信号转导途径：钙和 MAP 激酶信号途径对 BzATP 对 IFN 的影响的贡献。 - ¿最后，我们将检查转录因子对 IFN-¿ 的招募。通过染色质免疫沉淀法检测基因启动子，以测试我们的基本假设，即 P2X7 连接导致 ifnb1 启动子的转录因子占据增加。 P2X7 介导的信号传导对转录因子募集的修饰对增强免疫反应和骨生物学具有影响。两个关键组成部分是与 Paul Bertics 的密切合作以及指导委员会更正式的监督 Paul Bertics 在我感兴趣的领域拥有丰富的专业知识，并且在培训初级教师方面拥有出色的记录。除了 Bertics 博士之外，我的委员会还由其他三位非常支持和成功的医师科学家组成： 通过儿科，我获得了实验室空间、受保护的时间和初始启动资金 广泛的专业知识。威斯康星大学通过医学院和研究生院提供了大量的知识交流以及临床和基础科学继续教育的论坛。 相关性：巨噬细胞通过分泌细胞因子参与许多炎症过程，包括对感染和自身免疫性疾病的反应。我的研究建议研究组织损伤释放的称为“嘌呤”的物质如何调节炎症细胞因子 IFN-¿ 的产生。对炎症过程的更多了解可能最终导致更好的疾病治疗。