Investigating the neuroinflammatory role of RIP1 kinase

研究 RIP1 激酶的神经炎症作用

• 批准号: 8848334
• 负责人: JUNYING YUAN
• 金额: $ 33.71万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8848334
• 关键词: APP-PS1; Adverse effects; Affinity; Alzheimer' s Disease; Alzheimer' s disease model; Animal Model; Anti-Inflammatory Agents; Anti-Tumor Necrosis Factor Therapy; Anti-inflammatory; Antibodies; Arthritis; Aspirin; Attention; Behavior; Binding; Brain; Cardiovascular system; Caspase Inhibitor; Cell Death; Cells; Cerebrospinal Fluid; Cerebrum; Cessation of life; Chronic; Clinical Research; Clinical Trials; Collection; Data; Death Domain; Dementia; Deposition; Disease; Elderly; Encephalitis; Epidemiologic Studies; Etanercept; Event; FDA approved; Functional disorder; Gene Expression; Genetic; Goals; Health; Human; Immune; In Vitro; Incidence; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Knowledge; Late Onset Alzheimer Disease; Mediating; Mediator of activation protein; Memory; Microglia; Molecular; Molecular Profiling; Molecular Target; Mus; Network-based; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Onset of illness; Oral; Oral Administration; Outcome; Pathogenesis; Patients; Pharmaceutical Preparations; Phosphotransferases; Pilot Projects; Play; Process; Production; Reaction; Relative (related person); Reporting; Risk; Role; Safety; Senile Plaques; Signal Pathway; Signal Transduction; Spinal; System; TNF gene; TNFRSF1A gene; Testing; Transgenic Mice; Tryptophan 2,3 Dioxygenase; Tumor Necrosis Factor Receptor; aged; base; cognitive function; cost; cytokine; disorder control; improved; in vivo; inhibitor/antagonist; macrophage; mild cognitive impairment; mouse model; neuron loss; receptor; response; small molecule; tau phosphorylation

DESCRIPTION (provided by applicant): The goal of this proposal is to validate the role and explore the molecular mechanism of RIP1 kinase as a mediator of inflammatory response in Alzheimer's disease (AD), a devastating neurodegenerative disorder and the leading cause of dementia of the elderly. Chronic brain inflammation, characterized by the presence of an increased number of microglia and elevated levels of proinflammatory cytokines, is a hallmark of AD. Increased levels of cerebral spinal TNF? were found in patients with mild cognitive impairment (MCI) at risk to develop AD, suggesting that CNS inflammation is an early event during the pathogenesis of AD. The role of inflammation in the pathogenesis of AD was further highlighted by a recent network-based integrative analysis of a large collection of gene expression profiles from patients of late-onset Alzheimer's disease (LOAD) which discovered the immune/microglia system, including multiple TLR receptors and TNF?, as the molecular system most strongly associated with the pathophysiology of the LOAD. When activated, microglia may release proinflammatory cytokines to drive the chronic progression of AD by exacerbating A? deposition and neuronal death. Identification of the molecular targets in microglia that can be safely modulated to inhibit their inflammatory response may provide new options for the treatment of AD. However, there is a lack of knowledge about the neuroinflammatory mechanism that can be specifically and effectively modulated. We have developed a highly specific and potent inhibitor of RIP1 kinase, 7-Cl-O-Nec-1, a small molecule with excellent oral availability and safety profile, and highly CNS permeable. RIP1 kinase, a death- domain containing Ser/Thr kinase, has an established role in mediating multiple downstream signaling pathways downstream of TNFR1. We found that RIP1 kinase also plays an important role in mediating the production of TNF? by microglia induced by A? in vitro and in PSAPP transgenic mice in vivo which can be effectively inhibited by 7-Cl-O-Nec-1. Furthermore, oral administration of 7-Cl-O-Nec-1 led to the reduction of amyloid plaques and improved behavior and memory of B6.Cg-Tg(APPswe, PSEN1dE9) 85Dbo/J mice (PSAPP) mice, a model for AD. Our study suggests that RIP1 kinase is an important target for inhibiting neuroinflammatory response in AD. This proposal is to test this hypothesis and investigate the mechanism by which RIP1 kinase mediates neuroinflammatory responses in microglia. Specific Aim 1: Investigating the role and mechanism by which RIP1 kinase mediates inflammatory response in microglia activated by oligomeric A? by testing the possible involvement of MKK7 and TLR signaling as downstream mediators of RIP1 signaling. Specific Aim 2: Investigating the role and mechanism of p62 in A? mediated RIP1 kinase activation in microglia by testing the hypothesis that oligomerized p62 provides a platform for mediating RIP1 activation. Specific Aim 3: Genetic confirmation of the role of RIP1 kinase in mediating inflammatory response in AD transgenic mice using a RIP1 kinase dead knockin mouse line.

描述(申请人提供)：这项建议的目标是验证RIP1激酶作为阿尔茨海默病(AD)炎症反应介质的作用和分子机制，AD是一种毁灭性的神经退行性疾病，也是老年人痴呆症的主要原因。慢性脑炎是AD的一个特征，其特征是小胶质细胞数量增加和促炎细胞因子水平升高。脑脊液肿瘤坏死因子水平升高？在轻度认知障碍(MCI)患者中发现有发生AD的风险，提示中枢神经系统炎症是AD发病过程中的早期事件。最近对晚发性阿尔茨海默病(LOAD)患者的大量基因表达谱进行了基于网络的综合分析，进一步强调了炎症在AD发病机制中的作用。该分析发现，免疫/小胶质细胞系统，包括多个TLR受体和肿瘤坏死因子？，是与LOAD的病理生理最密切相关的分子系统。当被激活时，小胶质细胞可能会释放促炎细胞因子，通过加重A？沉积和神经元死亡。识别小胶质细胞中可安全调节以抑制其炎症反应的分子靶点，可能为AD的治疗提供新的选择。然而，对于可以被特异性和有效地调节的神经炎性机制缺乏了解。我们开发了一种高度特异和有效的RIP1激酶抑制剂7-Cl-O-NEC-1，这是一种具有极好的口服利用度和安全性且具有高度中枢神经系统通透性的小分子。RIP1是一个含有丝氨酸/苏氨酸的死亡结构域，在调节TNFR1下游的多条下游信号通路中起着重要的作用。我们发现RIP1激酶在介导肿瘤坏死因子的产生中也起重要作用。通过A？7-氯-O-NEC-1在体外和体内均能有效抑制PSAPP转基因小鼠的生长。此外，口服7-氯-O-NEC-1可使AD模型B6-Cg-TG(APPswe，PSEN1dE9)85Dbo/J小鼠(PSAPP)的淀粉样斑块减少，行为记忆能力改善。我们的研究表明，RIP1激酶是抑制AD神经炎性反应的重要靶点。这项提议是为了验证这一假说，并研究RIP1激酶介导小胶质细胞神经炎性反应的机制。具体目的1：探讨RIP1激酶在寡聚体A？激活的小胶质细胞炎症反应中的作用和机制。通过测试MKK7和TLR信号作为RIP1信号的下游介体的可能参与。具体目标2：探讨p62在A？通过验证寡聚p62为介导RIP1激活提供平台的假设，介导了小胶质细胞中RIP1激酶的激活。具体目的3：利用RIP1激酶死亡敲击小鼠系，从遗传学上证实RIP1激酶在AD转基因小鼠中介导炎性反应的作用。