Regulation of IFN-beta induction by P2X7 purinergic receptor signaling

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IFN产生的调节。作为一名医生科学家，我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。我近期的职业目标是发展一个可行的独立研究项目。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，以通过嘌呤能受体如P2 X7刺激免疫细胞。众所周知，ATP刺激P2 X7可增加内毒素（LPS）刺激的巨噬细胞产生炎性细胞因子（如IL-1？）。我们已经发现，用LPS和ATP类似物BzATP共刺激巨噬细胞大大增强了LPS对IFN-γ的诱导。I型IFN在先天性和适应性免疫的各个方面发挥重要作用。作为IFN-γ和嘌呤能受体信号转导与细菌性脓毒症和骨形态发生有关，因此了解P2 X7如何调节IFN-γ诱导的机制至关重要。为了开始剖析P2 X7如何发挥其作用的机制，提出了一种多管齐下的方法：我们将研究P2 X7连接对不同感染性刺激（例如LPS，dsRNA等）诱导IFN-γ的作用。以确定P2 X7增强是否是LPS特异性的。已知P2 X7连接启动多种信号转导途径：将分析钙和MAP-激酶信号转导途径对BzATP对IFN-γ诱导的作用的贡献。最后，我们将检查招聘的转录因子的IFN-γ基因启动子染色质免疫沉淀测试我们的基本假设，P2 X7连接导致增加的转录因子占用的ifnb 1启动子。通过P2 X7介导的信号传导对转录因子募集的修饰对于增强免疫应答和骨生物学具有分支。职业发展计划：两个关键组成部分是与Paul Bertics的密切合作和由指导委员会进行更正式的监督。Paul Bertics在我感兴趣的领域拥有丰富的专业知识，在培训初级教师方面有着出色的记录。除了伯蒂奇博士，我的委员会由另外三位非常支持和成功的医生科学家组成。研究环境：通过儿科系，我获得了实验室空间、受保护的时间和初始启动资金。通过医学和研究生院，威斯康星州大学的专业知识的广度是巨大的。我需要的任何设备或技术援助都可以提供。在临床和基础科学方面，有大量的学术交流和继续教育论坛。 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。我的研究旨在研究组织损伤释放的物质“嘌呤”如何调节巨噬细胞产生炎症细胞因子IFN-γ。更好地了解炎症过程可能最终导致更好地治疗疾病。

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.