Arginase-1 signaling after neonatal stroke

新生儿中风后精氨酸酶 1 信号转导

基本信息

批准号：
10664501
负责人：
Jana Krystofova Mike
金额：
$ 19.72万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-02-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10664501
关键词：
3-Dimensional Address Affect Age Apoptotic Area Arginine Biological Assay Brain Brain Hypoxia-Ischemia Brain region Cells Child Cicatrix Coagulation Process Collagen Type I Common carotid artery Complex Cytoskeleton Data Data Analyses Defect Ensure Exhibits Exposure to Extracellular Matrix Extracellular Matrix Proteins Funding Gene Expression Profile Goals Growth Histologic Hypoxia Hypoxic-Ischemic Brain Injury Immune response Individual Inflammation Ingestion Injury Ischemia Knowledge Laboratories Learning Mediating Mentors Methodology Methods Microglia Modeling Molecular Mus Nerve Regeneration Neuroglia Neurons Organ Outcome Pathway interactions Performance Phagocytes Physicians Polyamines Procedures Process Production Proline Proteins RNA Research Research Project Summaries Role Scientist Signal Pathway Signal Transduction Site Structure Techniques Testing Therapeutic Time Tissues Translating Tubular formation Use of New Techniques Western Blotting animal care arginase career career development cell type disability experimental study improved in vivo inhibitor migration multiple omics natural hypothermia neonatal brain neonatal hypoxic-ischemic brain injury neonatal stroke neural neurodevelopment neuroimmunology neuroinflammation neuropathology neuroprotection novel post stroke regenerative regenerative tissue repaired response spatiotemporal stem cells timeline tissue regeneration tissue repair tissue-repair responses transcriptome

项目摘要

PROJECT SUMMARY This research plan is based on a strong scientific premise that hypoxic-ischemic (HI) brain injury induces brain arginase-1 (ARG-1) to exhibit selected neuroprotective functions, such as efferocytosis and regenerative scar formation. In our preliminary studies, we detected spatiotemporal changes in ARG1 expression and activity as a result of neonatal HI. We have shown ARG1 localized mostly in microglia at the injury site early after injury performing efferocytosis and persisted in the injury core at later timepoints in the area of the tissue scar. ARG1 inhibition decreased ARG1 efferocytic function and worsened histological outcomes. While ARG1 involvement in efferocytosis and scar formation in other organs is well documented, ARG1-dependent mechanisms of efferocytosis and scar formation in the neonatal brain after HI are unknown. I hypothesize that ARG-1 regulates efferocytosis by providing polyamines for cytoskeleton assembly, and efficient efferocytosis is a crucial process for regenerative scar formation where ARG1 provides proline for extracellular matrix formation. Using in vivo the Vannucci procedure (common carotid artery coagulation followed by exposure to hypoxia in P9 mice) to induce HI, I will test my hypothesis in the following Specific aims: I will define whether HI induces polyamine pathway in ARG1 microglia, and whether ARG1 inhibition translates to defects in cytoskeleton assembly and performance of efferocytosis (Aim 1). I will characterize how ARG1 signaling alters cell composition in the scar and production of the extracellular matrix (Aim 2). I will determine whether efferocytosis is necessary for scar formation, if changes in ARG1 signaling alter local immune response in the injury core and whether this impacts migration of progenitor cells to the scar and tissue remodeling (Aim 3). The aims will be conducted using novel techniques, such as TRAPseq and spatial seqFISH that will significantly improve our understanding of processes in individual cells and cellular transcriptome in 3D. The proposed project will significantly improve our understanding of arginase-1 pathway, efferocytosis and tissue regeneration as a modifier of brain hypoxic-ischemic injury. This project will also significantly advance my scientific growth through learning besides the basics of neuroimmunology, the multiomics approaches and advanced data analysis necessary for my independent research career. Dr. Ferriero and I selected outstanding mentors, that together with coursework and research plans are aligned to address my specific knowledge gaps to ensure my career development as an independently funded physician scientist and to apply for an R01 at the end of this proposal. The proposed experiments and timeline are within my capabilities and the capabilities of the laboratory, animal care, and UCSF facilities.

项目概要该研究计划基于一个强有力的科学前提，即缺氧缺血（HI）脑损伤会诱发脑损伤精氨酸酶-1 (ARG-1) 表现出选定的神经保护功能，例如胞吞作用和再生疤痕形成。在我们的初步研究中，我们检测到 ARG1 表达和活性的时空变化：新生儿 HI 的结果。我们已经表明 ARG1 在损伤后早期主要定位于损伤部位的小胶质细胞中执行胞吞作用并在组织疤痕区域的稍后时间点持续存在于损伤核心中。 ARG1 抑制降低了 ARG1 细胞功能并恶化了组织学结果。当ARG1参与时在其他器官的胞吞作用和疤痕形成中，ARG1 依赖性机制已得到充分记录。 HI 后新生儿大脑中的胞吞作用和疤痕形成尚不清楚。我假设 ARG-1 调节通过为细胞骨架组装提供多胺来进行胞吞作用，而有效的胞吞作用是一个关键过程用于再生性疤痕形成，其中 ARG1 为细胞外基质形成提供脯氨酸。在体内使用 Vannucci 程序（颈总动脉凝固，然后 P9 小鼠暴露于缺氧）以诱导 HI，我将在以下方面检验我的假设具体目标：我将定义 HI 是否会诱导多胺途径 ARG1 小胶质细胞，以及 ARG1 抑制是否会导致细胞骨架组装和性能缺陷胞吞作用（目标 1）。我将描述 ARG1 信号传导如何改变疤痕和生产中的细胞组成细胞外基质（目标 2）。我将确定胞吞作用对于疤痕形成是否是必要的，如果 ARG1 信号的变化会改变损伤核心的局部免疫反应，以及这是否会影响祖细胞对疤痕和组织重塑的影响（目标 3）。这些目标将使用新技术来实现，例如 TRAPseq 和空间 seqFISH 将显着提高我们对个体过程的理解 3D 细胞和细胞转录组。拟议的项目将显着提高我们对精氨酸酶-1 途径、胞吞作用和组织再生作为脑缺氧缺血性损伤的调节剂。这除了基础知识之外，该项目还将通过学习显着促进我的科学成长神经免疫学、多组学方法和高级数据分析是我的独立研究所必需的研究生涯。 Ferriero 博士和我挑选了杰出的导师，以及课程作业和研究计划旨在解决我的具体知识差距，以确保我作为独立的职业发展资助医师科学家并在本提案末尾申请 R01。所提出的实验和时间表在我的能力范围内以及实验室、动物护理和加州大学旧金山分校设施的能力范围内。