Nervous system control of intestinal host defense mediated by TFEB

项目摘要 在美国，肠道感染和炎症性肠病（IBD）引起很高的发病率， 大部分医疗资源。对肠道健康中宿主微生物动态的了解有限， 疾病是旨在增强免疫力和减少炎症的治疗的障碍。有一个很大的联合国- 满足了对肠上皮在与肠道细菌通信中的作用的基本理解的需要， teria。该项目的长期目标是揭示体内参与的调节途径， 在肠上皮中诱导宿主防御基因。本提案的总体目标是 鉴定调控PLC-PKD-TFEB的上游机制（C. elegans）路径- 我们在前一个资助期确定的方式，以及它驱动基因的下游机制， 诱导中心假设是胆碱能神经系统控制PLC-1/DKF-1/HLH-30 这是一个重要的信号通路，它诱导一个转录因子级联反应，这是重要的宿主防御肠道。的 提出这项研究的理由是，完全了解神经系统如何与 整个PLC-1/DKF-1/HLH-30途径对于基本了解宿主防御至关重要，并且 更好地掌握其工作原理是必要的，以利用其作为炎症或炎症的治疗靶点的潜力。 感染为了检验中心假设，我们提出了以下具体目标：1）确定 神经系统在HLH-30/TFEB的激活，通过定义神经元检测感染，目标远端 2）确定DKF-1/PKD和HLH之间的联系- 30/TFEB，通过反向遗传学，HLH-30/TFEB磷酸化分析，并定义蛋白质-蛋白质相互作用;和 3)确定下游转录因子在介导HLH-30/TFEB功能中的作用，通过肠- 特异性功能丧失和转录谱分析。拟议研究的预期贡献是 确定胆碱能神经系统在控制PLC-PKD-TFEB通路及其下游的作用， 体内感染过程中流式转录因子网络。这一贡献是重要的，因为它将代表- 这为理解神经细胞调节上皮屏障的分子基础带来了飞跃。 系统，并将使未来的治疗，操纵新兴的毒蕈碱- 转录控制网络这项研究是非常创新的，因为PLC-PKD- TFEB通路尚未在任何背景下进行研究，因此所提出的研究极具创新性。 这里提出的研究代表了一个新的和实质性的脱离现状，转移重点 新发现的PLC-PKD-TFEB介导的新的神经系统-肠道信号传导轴 通路所提出的工作克服了许多障碍，全面了解主机防御， 并有很大的潜力，以加快创新疗法的发展，针对新兴的蝇， rinic控制的转录网络来治疗感染性或炎性疾病。

PROJECT SUMMARY In the U.S., enteric infections and inflammatory bowel diseases (IBD) cause great morbidity and consume a large portion of health-care resources. Limited knowledge of the host-microbe dynamic in intestinal health and disease is a barrier for therapies aimed at boosting immunity and reducing inflammation. There is a large un- met need for fundamental understanding of the role of the intestinal epithelium in communication with gut bac- teria. The long-term goal of this project is to uncover regulatory pathways that are engaged in vivo for the in- duction of host defense genes in the intestinal epithelium. The overall objective of the present proposal is to identify upstream mechanisms that regulate the PLC-PKD-TFEB (PLC-1/DKF-1/HLH-30 in C. elegans) path- way that we identified in the previous funding period, and downstream mechanisms by which it drives gene induction. The central hypothesis is that the cholinergic nervous system controls the PLC-1/DKF-1/HLH-30 pathway, which induces a transcription factor cascade that is important for host defense in the intestine. The rationale for the proposed research is that complete understanding of how the nervous system interacts with the entire PLC-1/DKF-1/HLH-30 pathway is essential for fundamental understanding of host defense, and that a better grasp of its workings is necessary to harness its potential as a therapeutic target for inflammation or infection. To test the central hypothesis, we propose the following specific aims: 1) Determine the role of the nervous system in the activation of HLH-30/TFEB, by defining neurons that detect infection, the target distal tissues, and pathogen genes necessary for detection; 2) Define the link between DKF-1/PKD and HLH- 30/TFEB, by reverse genetics, HLH-30/TFEB phosphoanalysis, and defining protein-protein interactions; and 3) Determine the role of downstream transcription factors in mediating HLH-30/TFEB function, by intestine- specific loss of function and transcriptional profiling. The expected contribution of the proposed research is to define the role of the cholinergic nervous system in controlling the PLC-PKD-TFEB pathway and its down- stream transcription factor network during infection in vivo. This contribution is significant because it will repre- sent a leap forward for understanding the molecular basis of regulation of the epithelial barrier by the nervous system, and will enable the future development of therapies that manipulate the emerging muscarinic- controlled transcriptional network. The proposed research is extremely innovative because the PLC-PKD- TFEB pathway has not been studied in any context, and thus the proposed research is extremely innovative. The research proposed here represents a new and substantive departure from the status quo, by shifting focus to a new nervous system-intestinal signaling axis that is mediated by the newly uncovered PLC-PKD-TFEB pathway. The proposed work overcomes many barriers to the comprehensive understanding of host defense, and has great potential to accelerate the development of innovative therapies that target the emerging musca- rinic-controlled transcriptional network to treat infectious or inflammatory diseases.