Cell Specific RIPK3 signaling after traumatic brain injury in mice

小鼠脑外伤后细胞特异性 RIPK3 信号转导

基本信息

批准号：
10377444
负责人：
MICHAEL J WHALEN
金额：
$ 42.43万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10377444
关键词：
Acute Aging Apoptosis Astrocytes Behavior Bone Marrow Brain Brain Injuries CASP8 gene CD95 Antigens Cell Death Cell Fraction Cells Cessation of life Chimera organism Chronic Chronic Phase Clinical Clinical Trials Cognitive deficits Cortical Contusions Cre lox recombination system Data Detergents Disease Down-Regulation Endothelial Cells Endothelium Etiology Event Experimental Models Fluorescence-Activated Cell Sorting Gene Expression Genetic Goals HMGB1 gene Hippocampus (Brain)Histology Human Immune Immunoprecipitation Impairment Inflammasome Inflammation Inflammation Mediators Inflammatory Injury Interleukin-1 beta Knock-out Knockout Mice Lead Lesion Link MAP Kinase Gene MAP3K7 gene Mediating Methodology Microglia Modeling Molecular Motor Mus Necrosis Nerve Degeneration Neurodegenerative Disorders Neurologic Dysfunctions Neurological outcome Neuronal Injury Neurons Outcome Pathway interactions Peripheral Phosphorylation Phosphotransferases Population Protein Kinase Protein-Serine-Threonine Kinases Proteins Publishing Quality of life RIPK1 gene RIPK3 gene Reagent Regulation Resolution Role Signal Transduction Survivors Systemic disease TBI Patients TBK1 gene TLR4 gene TNF gene TNFRSF1A gene TRADD gene Tamoxifen Testing Traumatic Brain Injury Tumor Necrosis Factor Receptor Tumor-infiltrating immune cells Ubiquitination Up-Regulation Western Blotting Work age related neurodegeneration antagonist brain cell brain tissue cell type cognitive function controlled cortical impact experimental study functional outcomes genetic regulatory protein grasp improved improved outcome inhibitor injured macrophage morris water maze motor deficit multicatalytic endopeptidase complex neuron loss new therapeutic target novel object recognition p38 Mitogen Activated Protein Kinase proteostasis response stroke model therapeutic target tool

项目摘要

Receptor interacting protein kinases (RIPK)-1 and -3 are serine-threonine kinases that regulate apoptosis, necrosis, and inflammation. RIPKs mediate programmed necrosis via RIPK1-RIPK3-MLKL necrosome assembly, and apoptosis via the ripoptosome involving FLIP, caspase-8, TRADD, and RIPK3, among others. RIPK1 inhibitors are currently in human clinical trials for several systemic diseases. In contrast, RIPK3 has been less studied in part because clinically acceptable RIPK3 inhibitors are not yet available. Building on our prior studies showing improved neurological outcome in mice deficient in TNF and Fas receptor (upstream activators of RIPK3), we used genetic tools to interrogate a possible role for RIPK3 in a mouse controlled cortical impact (CCI) model. Mice lacking RIPK3 had improved motor and cognitive function after CCI, and greater protection in cognitive function tests than RIPK1 kinase dead or MLKL knockout mice, suggesting a unique role for RIPK3- independent of necroptosis because acute neuronal cell death or lesion volume was not reduced by RIPK3 knockout. Using immunopanning to isolate specific brain cell populations, we found the highest levels of RIPK3 expression in endothelium and immune cells. RIPK3 KO mice had reduced HMGB1 release after CCI, reduced interleukin-1 beta processing in brain tissue and endothelial cells, and maintained K48 ubiquitination of neuronal proteins and brain TBK1 levels- mechanisms that have previously been shown to modulate outcome after CCI and contribute to neurodegeneration in aging-related neurodegenerative diseases. We hypothesize that RIPK3 signaling in endothelial and immune cells induces neurological dysfunction after cerebral contusion by activating multiple acute inflammation pathways and is a potential therapeutic target to improve outcome. To test this hypothesis we propose three specific aims: Aim 1, to identify upstream mechanisms regulating RIPK3 activation in specific brain cell types after CCI in mice, using immunopanning and FACS to isolate specific brain cell types; Aim 2, to define cell-specific functional roles for RIPK3 in brain vs. peripheral immune cells using bone marrow chimeras and inducible/conditional RIPK3 knockout mice; and Aim 3, to identify mechanisms downstream of RIPK3 that might contribute to neurodegeneration in the chronic period after TBI.

受体相互作用的蛋白激酶（RIPK）-1和-3是调节凋亡的丝氨酸 - 硫代激酶坏死和炎症。 RIPK通过RIPK1-RIPK3-MLKL坏死体介导编程的坏死通过涉及翻转，caspase-8，tradd和ripk3等的Ripopoptosoms组的组装和凋亡。 RIPK1抑制剂目前正在接受几种全身性疾病的人类临床试验。相比之下，ripk3具有较少研究的部分是因为尚无临床可接受的RIPK3抑制剂。建立在我们的基础上先前的研究表明，TNF和FAS受体缺乏的小鼠的神经系统结局改善（上游 RIPK3的激活剂，我们使用遗传工具来询问RIPK3在鼠标受控中的可能作用皮质影响（CCI）模型。缺乏RIPK3的小鼠在CCI后的运动和认知功能提高了，并且与RIPK1激酶死亡或MLKL敲除小鼠相比，认知功能测试的保护更大 RIPK3的独特作用 - 独立于坏死病，因为急性神经元细胞死亡或病变体积不是由RIPK3淘汰赛减少。使用免疫人员分离特定的脑细胞群体，我们发现内皮和免疫细胞中RIPK3表达的最高水平。 RIPK3 KO小鼠减少了HMGB1 CCI后释放，减少了脑组织和内皮细胞中的白介素-1β处理，并保持 K48神经元蛋白和脑TBK1水平的泛素化 - 先前已显示的机制调节CCI后的结果，并有助于与衰老相关的神经退行性的神经退行性。疾病。我们假设内皮细胞和免疫细胞中的RIPK3信号传导会诱导神经系统通过激活多个急性炎症途径，大脑挫伤后功能障碍治疗目标以改善预后。为了检验这一假设，我们提出了三个具体目标：目标1，使用小鼠CCI后特定脑细胞类型中调节RIPK3激活的上游机制，使用免疫剂和FACS分离特定的脑细胞类型； AIM 2，定义细胞特异性功能作用 RIPK3使用骨髓嵌合体和诱导/条件RIPK3中的脑与周围免疫细胞淘汰小鼠；和目标3，以确定RIPK3下游的机制可能有助于 TBI之后的慢性时期神经变性。