To the edge of necroptosis and back

到坏死性凋亡的边缘和背部

• 批准号: 9318899
• 负责人: DOUGLAS R GREEN
• 金额: $ 42.63万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-06 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9318899
• 关键词: Affect; Alpha Cell; Antigens; Apoptosis; Atherosclerosis; Back; Binding; Biochemical; Biological; CASP8 gene; Caspase; Cause of Death; Cell Death; Cell Survival; Cell membrane; Cell surface; Cells; Cessation of life; Complex; Cytokinesis; DNA Damage; Elements; Engineering; Event; Exocytosis; Gene Expression; Gene Proteins; Genetic; Goals; Image; Interferons; Ligation; Malignant Neoplasms; Mediating; Membrane; Mitochondria; Morphology; Multivesicular Body; Mutation; Necrosis; Nucleic Acids; Pathologic; Pathway interactions; Phosphatidylserines; Phosphorylation; Phosphotransferases; Play; Process; Production; Proliferating; Protein Kinase; Proteins; RIPK1 gene; Receptor Activation; Regulation; Reperfusion Injury; Research; Resuscitation; Role; Rupture; Sepsis; Shapes; Signal Transduction; Staining method; Stains; Stimulus; System; TNFRSF1A gene; Therapeutic; Viral Proteins; adaptive immunity; annexin A5; base; inhibitor/antagonist; interest; mixed lineage kinase 3; nonalcoholic steatohepatitis; novel; prevent; protein expression; receptor; receptor function; response; sensor; trafficking

Abstract Necroptosis is a form of cell death, inhibited by the FADD-caspase-8-FLIP complex, in which the interaction of two protein kinases, RIPK1 and RIPK3, interact to promote or inhibit necrotic death of a cell. Activated RIPK3 phosphorylates the pseudokinase MLKL, which mediates necroptosis through action at the plasma membrane. We have probed the late events that surround MLKL activation and have found that prior to loss of plasma membrane integrity, cell death is reversible. We have implicated the ESCRT III pathway, which functions in endosomal trafficking, cytokinesis, and exocytosis, in counter-acting the membrane effects of active MLKL. This results in the release of “bubbles” of damaged plasma membrane from the cell surface, sustaining cell survival. Our central hypothesis, upon which this application is based, is that necroptosis proceeds via the activation of MLKL and its action at the plasma membrane, and the regulation of this late process allows other events, such as protein expression induced by the necroptotic stimulus to manifest and thereby impact on the effects of the dying cell on the body. Based on this hypothesis, we will ask: 1. How do the late events in necroptosis regulate cell death and survival? Here we will identify the components of ESCRT III responsible for bubble formation after MLKL activation, and their role in “resuscitation” when active MLKL is subsequently disrupted. We will determine how ESCRT III functions to inhibit necroptosis in response to inducers without caspase inhibition, and explore if RIPK3 activity, independently of MLKL, may regulate the ESCRT III pathway. 2. Are there non-canonical pathways to MLKL activation and necroptosis? While several inducers of necroptosis have been identified, we propose that there are others that are less explored. We have identified DNA damage as an inducer of necroptosis, and will explore how this occurs. Further, we have found agents that appear to activate MLKL and necroptosis independently of RIPK3, and will determine how this non-canonical activation occurs. 3. What are the consequences of the late events in necroptosis for other cells? Cells undergoing necroptosis affect surrounding cells. Here we will ask how ESCRT III impacts on engulfment of the dying cell, how the delay in death supports gene expression and protein production, and allows engagement of adaptive immunity to antigens associated with the dying cell. Completion of our goals will advance our fundamental understanding of how necroptosis is regulated, and how it can be exploited in therapies directed to cancer and other pathological conditions.

摘要 坏死性凋亡是细胞死亡的一种形式，由FADD-胱天蛋白酶-8-FLIP复合物抑制，其中FADD-胱天蛋白酶-8-FLIP复合物与FADD-胱天蛋白酶-8-FLIP复合物的相互作用抑制细胞死亡。 两种蛋白激酶RIPK 1和RIPK 3相互作用以促进或抑制细胞的坏死性死亡。激活RIPK 3 磷酸化假激酶MLKL，其通过作用于质膜介导坏死性凋亡。 我们已经探测了围绕MLKL激活的晚期事件，并发现在血浆损失之前， 细胞膜完整，细胞死亡是可逆的。我们已经暗示了ESCRT III途径，其在 内体运输、胞质分裂和胞吐作用，抵消活性MLKL的膜效应。 这导致受损质膜的“气泡”从细胞表面释放，维持细胞存活。 生存我们的中心假设，即本申请所基于的假设，是坏死性凋亡通过 MLKL的激活及其在质膜上的作用，以及对这一晚期过程的调节允许其他 事件，如由坏死性凋亡刺激诱导的蛋白质表达，以显现并从而影响细胞的功能。 垂死细胞对身体的影响。基于这一假设，我们将问：1。最近发生的事件 坏死性凋亡调节细胞死亡和存活？在这里，我们将确定ESCRT III的组成部分 负责MLKL活化后的气泡形成，以及当活性MLKL被激活时它们在“复苏”中的作用。 随后被打乱。我们将确定ESCRT III如何发挥作用，以抑制坏死性凋亡， 诱导剂无半胱天冬酶抑制，并探讨RIPK 3活性，独立于MLKL，是否可以调节 ESCRT III途径。2. MLKL激活和坏死性凋亡是否存在非经典途径？而 坏死性凋亡的几种诱导物已经被鉴定，我们认为还有其他的较少被探索。 我们已经确定DNA损伤是坏死性凋亡的诱导物，并将探讨这是如何发生的。我们还 已经发现了似乎独立于RIPK 3激活MLKL和坏死性凋亡的药物，并将确定 这种非典型的激活是如何发生的3.坏死性凋亡晚期事件的后果是什么 其他细胞？经历坏死性凋亡的细胞会影响周围的细胞。在这里，我们将问如何ESCRT III 死亡的延迟如何支持基因表达和蛋白质 在某些实施方案中，免疫原性抗体可以产生，并且允许对与垂死细胞相关的抗原的适应性免疫接合。 完成我们的目标将推进我们对坏死性凋亡是如何调节的基本理解，以及如何 它可以用于针对癌症和其它病理状况的治疗。