Role of RIPK2 in the neuroinflammatory response to ischemic stroke

RIPK2 在缺血性卒中神经炎症反应中的作用

基本信息

批准号：
10680081
负责人：
Eduardo Jesus Candelario-Jalil
金额：
$ 40.89万
依托单位：
UNIVERSITY OF FLORIDA
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：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10680081
关键词：
Ablation Acute Address Adult Alteplase Animal Model Animals Apoptosis Attenuated Basic Science Benzamides Brain Brain Injuries Brain Ischemia CRISPR/Cas technology Cause of Death Cell Death Cell Lineage Cells Chronic Coagulation Process Data Dementia Dendritic Cells Diabetes Mellitus Dose Enterobacteria phage P1 Cre recombinase Excision Exons Financial Hardship Flow Cytometry Foundations Goals Healthcare Systems Hour Immune Immunoassay Impairment Industry Infarction Infiltration Inflammation Inflammation Mediators Inflammatory Injury Intervention Invaded Ischemia Ischemic Brain Injury Ischemic Penumbra Ischemic Stroke Knockout Mice Knowledge Label LoxP-flanked allele Macrophage Magnetic Resonance Imaging Measures Mechanics Microglia Mission Modeling Molecular Mouse Strains Mus Myelogenous Myeloid Cells National Institute of Neurological Disorders and Stroke Neurologic Neurologic Deficit Neurons Neuroprotective Agents Obesity Outcome Pathology Pathway interactions Patients Pattern Peripheral Phase Phenotype Phosphotransferases Process Production Protac Proteins Public Health RIPK2 gene Receptor Inhibition Receptor-Interacting Serine/Threonine Protein Kinase 2 Reperfusion Therapy Research Rodent Role Sensorimotor functions Signal Transduction Site Stroke Tamoxifen Technology Therapeutic Thrombectomy Time Tracer Western Blotting Wild Type Mouse aged behavioral outcome blood-brain barrier permeabilization cell type chemokine clinically relevant cognitive function comorbidity cytokine disability functional outcomes genome editing glial activation immune cell infiltrate improved improved outcome in vivo inhibitor innovation kinase inhibitor monocyte nano-string nervous system disorder neuroinflammation neurological recovery neuronal survival neuroprotection new therapeutic target novel novel drug class pharmacologic post stroke recruit response sex stroke model stroke outcome stroke patient stroke therapy success thrombolysis tool

Ablation Acute Address Adult Alteplase Animal Model Animals Apoptosis Attenuated Basic Science Benzamides Brain Brain Injuries Brain Ischemia CRISPR/Cas technology Cause of Death Cell Death Cell Lineage Cells Chronic Coagulation Process Data Dementia Dendritic Cells Diabetes Mellitus Dose Enterobacteria phage P1 Cre recombinase Excision Exons Financial Hardship Flow Cytometry Foundations Goals Healthcare Systems Hour Immune Immunoassay Impairment Industry Infarction Infiltration Inflammation Inflammation Mediators Inflammatory Injury Intervention Invaded Ischemia Ischemic Brain Injury Ischemic Penumbra Ischemic Stroke Knockout Mice Knowledge Label LoxP-flanked allele Macrophage Magnetic Resonance Imaging Measures Mechanics Microglia Mission Modeling Molecular Mouse Strains Mus Myelogenous Myeloid Cells National Institute of Neurological Disorders and Stroke Neurologic Neurologic Deficit Neurons Neuroprotective Agents Obesity Outcome Pathology Pathway interactions Patients Pattern Peripheral Phase Phenotype Phosphotransferases Process Production Protac Proteins Public Health RIPK2 gene Receptor Inhibition Receptor-Interacting Serine/Threonine Protein Kinase 2 Reperfusion Therapy Research Rodent Role Sensorimotor functions Signal Transduction Site Stroke Tamoxifen Technology Therapeutic Thrombectomy Time Tracer Western Blotting Wild Type Mouse aged behavioral outcome blood-brain barrier permeabilization cell type chemokine clinically relevant cognitive function comorbidity cytokine disability functional outcomes genome editing glial activation immune cell infiltrate improved improved outcome in vivo inhibitor innovation kinase inhibitor monocyte nano-string nervous system disorder neuroinflammation neurological recovery neuronal survival neuroprotection new therapeutic target novel novel drug class pharmacologic post stroke recruit response sex stroke model stroke outcome stroke patient stroke therapy success thrombolysis tool

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项目摘要

Abstract Neuroinflammation after stroke significantly contributes to neuronal damage and neurological impairment. Delayed cell death in the ischemic penumbra is associated with glial activation and recruitment and infiltration of peripheral immune cells to the brain. This is triggered by the production of pro-inflammatory cytokines and chemokines, contributing to cell death and blood-brain barrier (BBB) permeability after stroke. Dying cells in the penumbra also release pro-inflammatory signals and damage-associated molecular patterns (DAMPs) that activate resident microglia toward a pro-inflammatory phenotype, thus further contributing to brain injury. Our overall goal is to reduce the spread of stroke damage by limiting neuroinflammation. Receptor interacting serine/threonine protein kinase 2 (RIPK2) is a critical mediator of inflammation via its activation of multiple pro-inflammatory and cell death pathways. Inhibition of RIPK2’s kinase activity abolishes its signaling to alleviate inflammatory conditions in the periphery. The role of RIPK2 in ischemic stroke remains unexplored; however, our pilot data shows a substantial reduction in infarct size and improvement in post-stroke functional outcomes, both acutely and long-term, in Ripk2 knockout (Ripk2-/-) mice compared to wild-type (Ripk2+/+) mice. We propose that RIPK2 is an essential initiator and propagator of pro-inflammatory pathways in ischemic stroke. Our main objective is to attenuate its activity and assess the specific role of RIPK2 in vivo as it relates to stroke pathology. We hypothesize that RIPK2 signaling is detrimental in ischemic stroke, and RIPK2 degradation/inhibition or selective ablation in myeloid cells will improve outcomes. Aim 1 will determine the neuroprotective effect of RIPK2 blockade after ischemia using a highly selective RIPK2 inhibitor and a proteolysis-targeting chimera (PROTAC) that specifically degrades RIPK2 in vivo. We will utilize aged mice of both sexes subjected to ischemic stroke and investigate the effects of RIPK2 blockade on infarct size and long-term behavioral outcomes. In Aim 2, we will determine the impact of RIPK2 blockade on stroke-induced neuroinflammation and investigate neuroprotection mechanisms. In Aim 3, we will dissect the cell-specific role of RIPK2 in the neuroinflammatory process after stroke by using Ripk2 floxed mice crossed with lines producing Cre recombinase in specific cell types. We will study the contribution of RIPK2 from myeloid- lineage cells and brain-resident microglia to stroke injury. This project will leverage our expertise and unique tools (Ripk2 floxed mice, PROTAC, and selective inhibitors) to understand the mechanisms of RIPK2-driven inflammation in the context of ischemic stroke. This research may lead to identifying RIPK2 as a new therapeutic target to block neuroinflammation and promote neuronal survival in the aftermath of an ischemic stroke.

抽象的中风后的神经炎症显着有助于神经元损伤和神经系统损害。缺血性半衰期延迟细胞死亡与神经胶质激活，募集和浸润有关周围免疫细胞到大脑。这是由促炎性细胞因子和趋化因子，导致中风后细胞死亡和血脑屏障（BBB）渗透性。垂死的细胞半月还释放促炎信号和与损伤相关的分子模式（湿）（湿）激活居民小胶质细胞向促炎的表型，从而进一步导致脑损伤。我们的总体目标是通过限制神经炎症来减少中风损伤的传播。受体相互作用丝氨酸/苏氨酸蛋白激酶2（RIPK2）是通过其炎症的关键介体多种促炎和细胞死亡途径的激活。 RIPK2激酶活性的抑制消除了它的信号传导以减轻周围的炎症状况。 RIPK2在缺血性中风中的作用意外;但是，我们的飞行员数据显示，梗塞大小和冲程后的改善大大降低了与野生型相比（RIPK2+/+）小鼠。我们建议RIPK2是促发炎途径的必不可少的引发剂和传播者缺血性中风。我们的主要目的是衰减其活动，并评估RIPK2在体内的特定作用与中风病理有关。我们假设RIPK2信号在缺血性中风有害，而RIPK2有害髓样细胞中的降解/抑制作用或选择性消融将改善预后。 AIM 1将使用高度选择性的缺血后RIPK2封锁的神经保护作用 RIPK2抑制剂和诱导蛋白水解的嵌合体（Protac），该嵌合体在体内特异性降解RIPK2。我们将利用两性的老年小鼠遭受缺血性中风并研究RIPK2封锁的影响梗塞大小和长期行为结果。在AIM 2中，我们将确定RIPK2封锁对中风引起的神经炎症并研究神经保护机制。在AIM 3中，我们将剖析 RIPK2在中风后通过使用RIPK2 Floxed小鼠与与之交叉的细胞特异性作用在特定细胞类型中产生CRE重组酶的线。我们将研究RIPK2的髓样贡献谱系细胞和脑居民小胶质细胞造成中风损伤。该项目将利用我们的专业知识和独特的工具（RIPK2 Floxed小鼠，Protac和选择性抑制剂）以了解RIPK2驱动的机制在缺血性中风的背景下发炎。这项研究可能导致将RIPK2识别为一种新的治疗在缺血性中风后阻止神经炎症并促进神经元存活的靶标。