Understanding genetic basis for resistance to TNF-induced lethal shock using new

使用新方法了解抵抗 TNF 诱导的致死性休克的遗传基础

• 批准号: 8073126
• 负责人: Alexander Poltorak
• 金额: $ 24.5万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-20 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8073126
• 关键词: Accounting; Acute; Address; Agonist; Apoptosis; Apoptotic; Attenuated; Biochemical; Cell Death; Cells; Cessation of life; Data; Defect; Dose; Employee Strikes; Equilibrium; Exhibits; Family member; Funding; Future; Galactosamine; Gene Expression; Genes; Genetic; Genetic Models; Genomics; Health; Hepatocyte; In Vitro; Individual; Inflammation; Inflammatory; Injury; Link; Lipopolysaccharides; Liver; Maps; Mediating; Mediator of activation protein; Modeling; Mouse Strains; Mus; Myocardial Infarction; Pathology; Pathway interactions; Peritoneal Macrophages; Phenotype; Phosphotransferases; Physiology; Reading; Receptor Signaling; Resistance; Septic Shock; Shock; Signal Transduction; Stroke; Therapeutic Intervention; Time; Toxic Shock Syndrome; Traumatic Brain Injury; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; attenuation; base; gene cloning; genetic analysis; genetic linkage analysis; in vitro Model; in vivo; in vivo Model; inhibitor/antagonist; insight; macrophage; mortality; novel; prevent; public health relevance; response; small molecule; trait; tumor

DESCRIPTION (provided by applicant): Mortality from toxic shock, caused by infective agents that trigger excessive synthesis of tumor necrosis factor (TNF) and other inflammatory molecules by macrophages, remains a major health problem. Here we describe for the first time resistance to lipopolysaccharide (LPS) and TNF-induced death in vivo of wild-derived mice of MSM strain, representing a new genetic model of resistance to toxic shock. The remarkable resistance of these mice to toxic shock is further emphasized by their lack of sensitivity to acute liver injury caused by the co-administration of TNF and D-galactosamine, a well-established in vivo model of endotoxic liver damage. In support of the in vivo resistance data, we show that primary MSM macrophages exhibit another phenotype in that they are resistant to TNF- or TLR-agonists-induced regulated necrosis ("necroptosis") but not apoptosis cell death in vitro. Based on these results, we hypothesize that the two traits are mechanistically linked and propose to examine the connection between necroptosis and toxic shock traits by means of genetic linkage analysis aimed at identification of the loci, which confer both traits. In further extension of the in vitro results, we discovered that TNF-mediated in vivo liver injury can be prevented by a specific inhibitor of a key necroptosis regulator, RIP1 kinase. In addition to cell death phenotype, we demonstrate that TNF synthesis by MSM macrophages is reduced and may also be influenced by RIP1 signaling in vivo. Based on our preliminary data, we suggest that RIP1 kinase is a mediator of acute toxic shock and may represent an important new target for therapeutic intervention. We propose to further characterize the scope of changes in in vitro MSM hepatocyte and macrophage responses to TLR agonists and TNF in order to better determine the cellular basis for MSM resistance to toxic shock. Overall, these studies may provide important new insights into mechanisms of both toxic shock in vivo and necroptotic RIP1 kinase signaling, which has also been implicated in other acute pathologies, such as stroke, myocardial infarction and brain trauma. PUBLIC HEALTH RELEVANCE: Mortality from toxic shock, caused by infective agents that trigger excessive synthesis of tumor necrosis factor (TNF) and other inflammatory molecules by macrophages remains a major health problem. In this proposal, using genetic analysis of resistance to toxic shock in the wild-derived mice, we will reveal novel mechanisms of control of inflammation and its magnitude.

描述（由申请人提供）：中毒性休克是由感染因子引发巨噬细胞过度合成肿瘤坏死因子（TNF）和其他炎症分子引起的死亡，仍然是一个主要的健康问题。在这里，我们首次描述了MSM品系野生小鼠体内对脂多糖（LPS）和TNF诱导的死亡的抵抗力，代表了一种新的抵抗中毒性休克的遗传模型。这些小鼠对中毒性休克的显着抵抗力进一步强调了它们对 TNF 和 D-半乳糖胺共同给药引起的急性肝损伤缺乏敏感性，这是一种完善的内毒素肝损伤体内模型。为了支持体内耐药性数据，我们表明初级 MSM 巨噬细胞表现出另一种表型，即它们对 TNF 或 TLR 激动剂诱导的调节性坏死（“坏死性凋亡”）有抵抗力，但在体外不抵抗细胞凋亡。基于这些结果，我们假设这两个性状在机械上相关，并建议通过旨在鉴定赋予这两个性状的基因座的遗传连锁分析来检查坏死性凋亡和中毒性休克性状之间的联系。在进一步扩展体外结果时，我们发现 TNF 介导的体内肝损伤可以通过关键坏死性凋亡调节因子 RIP1 激酶的特异性抑制剂来预防。除了细胞死亡表型外，我们还证明 MSM 巨噬细胞合成的 TNF 减少，并且也可能受到体内 RIP1 信号传导的影响。根据我们的初步数据，我们认为 RIP1 激酶是急性中毒性休克的介质，可能代表治疗干预的重要新靶点。我们建议进一步表征体外 MSM 肝细胞和巨噬细胞对 TLR 激动剂和 TNF 反应的变化范围，以便更好地确定 MSM 抗中毒性休克的细胞基础。总体而言，这些研究可能为体内中毒性休克和坏死性RIP1激酶信号传导机制提供重要的新见解，这些信号也与其他急性病理学有关，例如中风、心肌梗死和脑外伤。 公共卫生相关性：中毒性休克是由感染因子引起的死亡，这些感染因子会引发巨噬细胞过度合成肿瘤坏死因子 (TNF) 和其他炎症分子，这仍然是一个主要的健康问题。在这项提案中，我们将利用野生小鼠对中毒性休克抵抗力的遗传分析，揭示控制炎症及其严重程度的新机制。