Bacterial disruption of neuroimmune pathways in a transparent brain

项目概要/摘要 免疫系统的细胞、细菌和神经元在每一个细胞上都在不断地相互作用。 人体的表面。最近的研究表明，细菌已经进化出了一种机制， 参与神经系统，通常使用调节炎症的吞噬细胞。 神经炎症，或神经组织炎症，被认为有助于常见的 神经退行性疾病，包括阿尔茨海默病和帕金森病，虽然 神经炎症的基本机制仍不完全清楚。长期 这项新创新者奖提案的目的是确定 感染期间的神经炎症。创新是我们对前沿的适应 斑马鱼分子工具包（光遗传学、生物传感器、体内生物素化） 交互.有了这个系统，我们可以对宿主和病原体进行基因改造， 细菌实时入侵透明的斑马鱼大脑。作为病原体， 激活人类神经炎症的分枝杆菌物种（结核性脑膜炎，麻风病）， 在斑马鱼中。我们还开发了新的斑马鱼脑感染模型， 从脑膜炎患者中分离的病原体，以确定： 项目1：细菌是如何从血液中侵入大脑的？ 方法：明确脑血管侵袭和血管损伤的内皮机制。 项目2：宿主与病原体因素如何驱动灾难性的脑炎症？ 方法：体内吞噬细胞的光遗传（光激活）控制。 项目3：宿主与病原体因素如何损伤神经元？ 方法：使用生物传感器识别神经元损伤的感染机制。 为了回答这些问题，这个提议结合了免疫生物学和神经生物学 斑马鱼大脑感染模型的工具。利用斑马鱼脑部感染的实时成像， 以及荧光细菌、神经元和吞噬细胞的遗传修饰， 炎症和神经变性事件可以以前所未有的细节直接观察到。 这项工作的完成实现了两个目标。首先，它将提供所需的机械信息 在未充分研究的脑感染中介导损伤的宿主细胞和分子上， 对生活在贫困中的人的影响尤为严重。这将有助于实现开发新的 限制脑部炎症的疗法除了感染，感染的基本生物学机制 由感染揭示的神经炎症可能与许多非感染性 神经退行性病症的特征在于神经炎症。

Project Summary/Abstract Cells of the immune system, bacteria, and neurons are constantly interacting on every surface of the human body. Recent studies show that bacteria have evolved mechanisms to engage with the nervous system, often using the phagocytes that regulate inflammation. Neuroinflammation, or inflammation in neurological tissue, is thought to contribute to common neurodegenerative conditions, including Alzheimer's disease and Parkinson's disease, although the basic mechanisms of neuroinflammation remain incompletely understood. The long-term objective of this New Innovator Award proposal is to define the molecular mechanisms of neuroinflammation during infection. The innovation is our adaptation of the cutting-edge zebrafish molecular toolkit (optogenetics, biosensors, in vivo biotinylation) to host-pathogen interactions. With this system, we can genetically modify host and pathogen, then observe bacteria invading the transparent zebrafish brain in real-time. As pathogens, we use mycobacterial species that activate neuroinflammation in humans (TB meningitis, leprosy) and in zebrafish. We have also developed new zebrafish brain infection models, using human pathogens isolated from meningitis patients, in order to determine: Project 1: How do bacteria invade the brain from the blood? Approach: Define the endothelial mechanisms of brain invasion and vascular injury. Project 2: How do host versus pathogen factors drive catastrophic brain inflammation? Approach: Optogenetic (light-activated) control of phagocytes in vivo. Project 3: How do host versus pathogen factors injure neurons? Approach: Identifying infectious mechanisms of neuronal injury using biosensors. To answer these questions, this proposal combines immunobiology and neurobiology tools with brain infection models in zebrafish. Using live imaging of zebrafish brain infection, and genetic modification of fluorescent bacteria, neurons, and phagocytes, the earliest inflammatory and neurodegenerative events can be directly observed in unprecedented detail. Completion of this work fulfills two goals. First, it will provide needed mechanistic information on the host cells and molecules that mediate injury in understudied brain infections that disproportionately affect people living in poverty. This will support the goal of developing new therapies that limit brain inflammation. Beyond infection, the basic biological mechanisms of neuroinflammation, revealed by infection, will likely be relevant to the many non-infectious neurodegenerative conditions that are characterized by neuroinflammation.