The role of Fas-associated death domain in necroptosis in vivo

Fas相关死亡结构域在体内坏死性凋亡中的作用

• 批准号: 8295838
• 负责人: ASTAR WINOTO
• 金额: $ 37.68万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-02 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8295838
• 关键词: Address; Affinity Chromatography; Antibiotics; Antigens; Apoptosis; Apoptotic; Autoimmunity; Bacteria; Binding Proteins; Biochemical; Caspase; Cell Aging; Cell Death; Cell Death Signaling Process; Cells; Cessation of life; Co-Immunoprecipitations; Complex; Data; Death Domain; Dendritic Cells; Erythrocytes; Exhibits; Fas Signaling Pathway; Human; Immune; Immune response; Immune system; Immunity; Infection; Inflammation; Inflammatory; Influenza; Injection of therapeutic agent; Interphase Cell; Knock-out; Knockout Mice; Lead; Ligands; Lipopolysaccharides; Mass Spectrum Analysis; Mediating; Modeling; Mus; Mutant Strains Mice; Myeloproliferative disease; Necrosis; Parasites; Pathway interactions; Phenotype; Phosphotransferases; Physiological; Play; Predisposition; Proliferating; Proteins; RIPK3 gene; Resistance; Role; Running; Salmonella infections; Serum; Shapes; Signal Pathway; Signal Transduction; Stimulus; Syndrome; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; TLR4 gene; Testing; Time; Tissues; Toll-like receptors; Toxoplasma gondii; Toxoplasmosis; Transfection; Transgenic Mice; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Vaccines; Viral; Western Blotting; adapter protein; aged; caspase-8; cell age; cell type; cytokine; improved; in vivo; macrophage; member; mouse development; mouse model; neutrophil; pathogen; premature; programs; receptor; response; ubiquitin-protein ligase; Address; Affinity Chromatography; Antibiotics; Antigens; Apoptosis; Apoptotic; Autoimmunity; Bacteria; Binding Proteins; Biochemical; Caspase; Cell Aging; Cell Death; Cell Death Signaling Process; Cells; Cessation of life; Co-Immunoprecipitations; Complex; Data; Death Domain; Dendritic Cells; Erythrocytes; Exhibits; Fas Signaling Pathway; Human; Immune; Immune response; Immune system; Immunity; Infection; Inflammation; Inflammatory; Influenza; Injection of therapeutic agent; Interphase Cell; Knock-out; Knockout Mice; Lead; Ligands; Lipopolysaccharides; Mass Spectrum Analysis; Mediating; Modeling; Mus; Mutant Strains Mice; Myeloproliferative disease; Necrosis; Parasites; Pathway interactions; Phenotype; Phosphotransferases; Physiological; Play; Predisposition; Proliferating; Proteins; RIPK3 gene; Resistance; Role; Running; Salmonella infections; Serum; Shapes; Signal Pathway; Signal Transduction; Stimulus; Syndrome; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; TLR4 gene; Testing; Time; Tissues; Toll-like receptors; Toxoplasma gondii; Toxoplasmosis; Transfection; Transgenic Mice; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Vaccines; Viral; Western Blotting; adapter protein; aged; caspase-8; cell age; cell type; cytokine; improved; in vivo; macrophage; member; mouse development; mouse model; neutrophil; pathogen; premature; programs; receptor; response; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Cell death is an integral part of the immune responses to pathogenic infection, and different forms of cell death can have different immunological consequences. For example, apoptosis is thought to clear viral infected cells and to control run-away immune responses whereas pyroptosis of macrophages can result in the release of cytokines. The Tumor Necrosis Factor Receptor (TNF-R) family of death receptors can trigger apoptosis but also an alternative form of death called programmed necrosis or necroptosis. Necroptosis is dependent on the RIP1 and RIP3 kinases but little is known on its physiological role. To address this, we have generated three lines of tissue- specific FADD knockout mice. FADD is a known adapter protein for all the TNF-R death receptors and thus FADD-deficient cells are resistant to death receptor-mediated apoptosis. However, stimulation of FADD-deficient cells leads to death through necroptosis. This occurs in T-cell receptor stimulated T cells from T-cell specific-FADD knockout (tFADD-/-) mice, in lipopolysaccharide (LPS) stimulated dendritic cells (DCs) from DC-specific-FADD deficient (dcFADD-/-) mice and in LPS stimulated macrophages from macrophage-specific FADD deficient (mFADD-/-) mice. T cells from tFADD-/- mice are functionally defective due to premature necroptosis. However, FADD-deficient DCs are functionally normal and can secrete cytokines when stimulated. Analysis of dcFADD-/- mice showed that some of their phenotypes are surprisingly similar to DC-less mice with imbalance erythrocytes and myeloproliferative disease. At the same time, dcFADD-/- mice exhibit a modest increase of inflammation. In contrast to other DC-specific apoptosis-resistant mice, aged dcFADD-/- mice don't suffer from autoimmunity but they appear to have an enhanced immune system. We hypothesize that necroptosis is one of the strategies for innate immune cells to stimulate the immune system in cases when receptor-induced apoptosis is blocked. In dcFADD-/- mice, necroptotic DCs releasing inflammatory contents can lead to improved immunity against pathogenic infection. This hypothesis will be tested in aim 1. Using DC-specific FADD-/-/MyD88-/- and dcFADD-/-/RIP3-/- mice, we will address whether constitutive stimulation of DCs through TLRs leads to necroptosis and release of inflammatory contents. To examine the possible enhanced immunity of dcFADD-/- mice, they will be challenged with influenza and Toxoplasma gondii and studied in details. Whether necroptosis is required for immunity against these two pathogens will be assessed using RIP3-/- mice. In Aim 2, we will unravel the biochemical signaling pathways involving FADD in necroptosis in response to TLR stimulation. Successful completion of these aims will greatly enhance our understanding of the host- pathogen interaction and the physiological role of necroptosis. ! ! ! PUBLIC HEALTH RELEVANCE: Cell death, including apoptosis, pyroptosis, autophagic cell death and necrosis, is an integral part of development of the mouse/human immune system and/or responses to pathogenic infection. Members of the tumor necrosis factor (TNF) death receptors can initiate apoptosis and a new form of necrosis called necroptosis but the physiological role of necroptosis is poorly understood; we propose in this application to understand the role of necroptosis in infection by studying mouse models deficient in an adapter protein required for the TNF cell death signals.

描述（由申请人提供）：细胞死亡是对致病感染的免疫反应的组成部分，不同形式的细胞死亡可能会带来不同的免疫学后果。例如，凋亡被认为可以清除病毒感染的细胞并控制失控的免疫反应，而巨噬细胞的凋亡可能导致细胞因子释放。肿瘤坏死因子受体（TNF-R）死亡受体家族会引发凋亡，但也可以触发一种替代的死亡形式，称为程序坏死或坏死。坏死性依赖于RIP1和RIP3激酶，但其生理作用鲜为人知。为了解决这个问题，我们已经生成了三条组织特异性FADD敲除小鼠。 FADD是所有TNF-R死亡受体的已知衔接蛋白，因此缺陷FADD的细胞对死亡受体介导的凋亡具有抗性。但是，刺激缺陷细胞通过坏死病导致死亡。这发生在T细胞受体受刺激的T特异性FADD敲除（TFADD - / - ）小鼠中，脂多糖（LPS）刺激的树突状细胞（DCS）中的DC特异性-FADD缺陷（DCFADD-/ - - / - ）小鼠的脂多糖细胞（DCS）刺激了lps刺激的乳腺癌中的乳mic以上的小鼠。 TFADD - / - 小鼠的T细胞由于过早坏死而在功能上有缺陷。但是，缺乏FADD的DC在功能上是正常的，并且在刺激时可以分泌细胞因子。对DCFADD - / - 小鼠的分析表明，它们的某些表型与具有失衡红细胞和骨髓增生性疾病的无DC无小鼠非常相似。同时，DCFADD - / - 小鼠表现出适度的炎症增加。与其他DC特异性凋亡的小鼠相比，老化的DCFADD - / - 小鼠不会患有自身免疫性，但它们似乎具有增强的免疫系统。我们假设坏死性是在受体诱导的凋亡被阻断的情况下，先天免疫细胞刺激免疫系统的策略之一。在DCFADD - / - 小鼠中，释放炎症含量的坏死性DC可以改善针对致病感染的免疫力。该假设将在AIM 1中进行检验。使用DC特异性FADD - / - / - /MYD88 - / - 和DCFADD - / - / - /RIP3 - / - 小鼠，我们将通过TLRS通过TLRS来解决DCS刺激是否导致致命性和炎症含量的释放。为了检查DCFADD - / - 小鼠的可能增强的免疫力，它们将受到流感和弓形虫贡迪的挑战，并进行了详细研究。是否需要使用RIP3 - / - 小鼠评估对这两种病原体的免疫性需要的坏死性。在AIM 2中，我们将探讨涉及FADD在坏死刺激中涉及FADD的生化信号通路。这些目标的成功完成将大大增强我们对宿主病原体相互作用和坏死作用的理解。呢呢呢 公共卫生相关性：细胞死亡，包括凋亡，凋亡，自噬细胞死亡和坏死，是小鼠/人免疫系统发展的组成部分和/或对病原感染的反应。肿瘤坏死因子（TNF）死亡受体的成员可以引发细胞凋亡和一种称为坏死的新形式，但对坏死的生理作用知之甚少。我们在此应用中建议通过研究缺乏TNF细胞死亡信号所需的衔接蛋白中的小鼠模型来了解坏死性在感染中的作用。