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

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.

抽象的严重中风后的神经炎症会导致神经元损伤和神经功能障碍。缺血半暗带中的延迟细胞死亡与神经胶质细胞的激活以及神经胶质细胞的招募和浸润有关。这是由促炎细胞因子的产生触发的。趋化因子，导致中风后细胞死亡和血脑屏障（BBB）通透性。半影还释放促炎信号和损伤相关分子模式 (DAMP)，激活驻留的小胶质细胞，使其产生促炎表型，从而进一步导致脑损伤。总体目标是通过限制神经炎症来减少中风损伤的扩散。受体相互作用丝氨酸/苏氨酸蛋白激酶 2 (RIPK2) 通过其激活多种促炎症和细胞死亡途径。抑制 RIPK2 激酶活性可消除。 RIPK2 在缺血性中风中的作用仍然存在。尚未探索；然而，我们的试点数据显示梗塞面积大幅减少，中风后症状得到改善。与野生型相比，Ripk2 敲除 (Ripk2-/-) 小鼠的短期和长期功能结果 (Ripk2+/+) 小鼠中，RIPK2 是促炎症途径的重要启动子和传播子。我们的主要目标是减弱其活性并评估 RIPK2 在体内的具体作用。我们担心 RIPK2 信号传导会导致缺血性中风，并且 RIPK2 与中风病理学相关。骨髓细胞的降解/抑制或选择性消融将改善结果。目标 1 将使用高度选择性的方法确定缺血后 RIPK2 阻断的神经保护作用 RIPK2 抑制剂和一种在体内特异性降解 RIPK2 的蛋白水解靶向嵌合体 (PROTAC)。使用患有缺血性中风的两性老年小鼠并研究 RIPK2 阻断对在目标 2 中，我们将确定 RIPK2 阻断对梗死面积和长期行为结果的影响。在目标 3 中，我们将剖析中风引起的神经炎症并研究神经保护机制。 RIPK2在中风后神经炎症过程中的细胞特异性作用，通过使用Ripk2 floxed小鼠与我们将研究来自骨髓的 RIPK2 的贡献。该项目将利用我们的专业知识和独特的方法来治疗中风损伤。工具（Ripk2 floxed 小鼠、PROTAC 和选择性抑制剂）来了解 RIPK2 驱动的机制这项研究可能会导致将 RIPK2 确定为一种新的治疗方法。目标是阻断神经炎症并促进缺血性中风后神经元的存活。