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(申请人提供)：中毒性休克的死亡率，由感染性因素引起，引发巨噬细胞过度合成肿瘤坏死因子和其他炎症分子，仍然是一个主要的健康问题。在这里，我们首次描述了MSM品系野生小鼠对脂多糖(LPS)的抵抗力和肿瘤坏死因子诱导的体内死亡，代表了一种新的抵抗中毒性休克的遗传模式。这些小鼠对中毒性休克的显著抵抗力进一步得到强调，因为它们对由肿瘤坏死因子和D-氨基半乳糖联合注射引起的急性肝损伤缺乏敏感性，D-氨基半乳糖是一种成熟的体内内毒素肝损伤模型。为了支持体内耐药数据，我们发现原代MSM巨噬细胞表现出另一种表型，即它们对肿瘤坏死因子或TLR激动剂诱导的调节性坏死(“坏死下垂”)具有抵抗力，但不能在体外诱导凋亡细胞死亡。基于这些结果，我们假设这两个性状是机械连锁的，并建议通过遗传连锁分析的方法来检验坏死性下垂和中毒性休克性状之间的联系，目的是识别决定这两个性状的基因座。在体外结果的进一步扩展中，我们发现，关键的坏死下垂调节因子RIP1激酶的特定抑制剂可以预防肿瘤坏死因子介导的体内肝损伤。除了细胞死亡表型外，我们还证明了MSM巨噬细胞的肿瘤坏死因子合成减少，并可能在体内受到RIP1信号的影响。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，可能是治疗干预的一个重要的新靶点。我们建议进一步确定MSM对TLR激动剂和肿瘤坏死因子的体外反应的变化范围，以便更好地确定MSM抵抗中毒性休克的细胞学基础。总体而言，这些研究可能为体内中毒性休克和坏死性RIP1激酶信号转导机制提供重要的新见解，RIP1信号转导信号也与其他急性病理有关，如中风、心肌梗死和脑创伤。 公共卫生相关性：由感染性物质引起的中毒性休克死亡，巨噬细胞过度合成肿瘤坏死因子和其他炎性分子仍然是一个主要的健康问题。在这项建议中，利用野生小鼠对中毒性休克的抵抗力的遗传分析，我们将揭示控制炎症及其程度的新机制。