Cysteine Proteases in Apoptosis

半胱氨酸蛋白酶在细胞凋亡中的作用

• 批准号: 7781346
• 负责人: Emad S Alnemri
• 金额: $ 33.65万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-15 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7781346
• 关键词: Acne; Acute suppurative arthritis due to bacteria; Adaptor Signaling Protein; Affect; Apical; Apoptosis; Apoptotic; Aspartate; Biochemical; Biochemical Genetics; Biological; Caspase; Caspase-1; Cell Line; Cell physiology; Cells; Chronic; Chronic Disease; Colchicine; Complex; Cysteine Protease; Cytoskeleton; Data; Defect; Disease; Disease Pathway; Exhibits; Familial Mediterranean Fever; Family; Feedback; Genes; Genetic; Human; Inflammation; Inflammatory; Inflammatory Response; Interleukin-1; Interleukin-4; Lead; Mediating; Microtubules; Missense Mutation; Mus; Mutate; Mutation; Natural Immunity; Osteomyelitis; Pathway interactions; Physiological; Play; Process; Proteins; Pyoderma Gangrenosum; Research; Role; Signal Pathway; Stimulus; Symptoms; Syndrome; System; Therapeutic; base; clinical application; cytokine; design; insight; macrophage; marenostrin; member; procaspase-1; protein complex; public health relevance; reconstitution; research study; response

DESCRIPTION (provided by applicant): Caspase-1 is a member of a family of aspartate-specific cysteine proteases, known as the caspase family, involved in apoptosis and inflammation. Recent studies revealed that caspase-1 is activated within large protein complexes or assemblies called the inflammasomes. The inflammasomes are composed of several adaptor proteins that interact with caspase-1 and induce its oligomerization and activation. Preliminary evidence from the applicant's lab suggest that pyrin, a protein mutated in the auto- inflammatory disease Familial Mediterranean fever (FMF), plays a major role in inflammation by assembling an inflammasome complex with the adaptor protein ASC, and procaspase-1 leading to ASC oligomerization, caspase-1 activation and IL-1¿ processing. Pyrin also appears to play an important role in apoptosis as macrophages from pyrin-truncation mice exhibit a defect in apoptosis compared to macrophages from wildtype mice in response to IL-4 and LPS stimulation. Evidence suggests that the inflammatory and perhaps the apoptotic activities of pyrin are regulated by two cytoskeleton associated proteins known as PSTPIP1 and PSTPIP2. Missense mutations in these proteins are associated with two further auto-inflammatory diseases, known as pyogenic arthritis, pyoderma gangrenosum, acne syndrome (PAPA) and chronic multifocal osteomyelitis (cmo) syndrome, respectively. In this competing continuation application three specific aims are proposed to study the physiological role of pyrin in apoptosis and inflammation and the role of the cytoskeleton in these two pyrin pathways. In the first specific aim, the role of pyrin in caspase-1 activation will be investigated through detailed biochemical and biological studies in cell-based and cell-free reconstitution systems and human and mouse macrophage cell lines. In the second specific aim, experiments are proposed to characterize the interaction of PSTPIP1 and PSTPIP2 with pyrin and determine their ability to regulate caspase-1 activation in human and mouse cells. In the third specific aim, biological, biochemical and genetic approaches will be used to elucidate the mechanism by which pyrin regulates apoptosis. Understanding the interplay of the pyrin-mediated inflammatory and apoptotic responses and the mechanism of activation of caspase-1 by pyrin is likely to contribute important insights into the role of pyrin in innate immunity and apoptosis, with obvious potential clinical applications. Public Health Relevance Statement: Mutations in the genes encoding pyrin and the pyrin-associated protein PSTPIP1 are associated with severe autoinflammatory diseases in humans. This research will investigate the physiological function of pyrin to elucidate its role in normal cellular function and how disease-associated mutations alter its function. The results of this research will help in the design and discovery of effective therapeutics to treat autoinflammatory diseases.

描述（由适用提供）：caspase-1是大天冬氨酸特异性半胱氨酸蛋白的成员，称为caspase家族，参与细胞凋亡和炎症。最近的研究表明，在称为炎症的大蛋白质复合物或组件中激活caspase-1。炎症由几种与caspase-1相互作用的衔接蛋白组成，并诱导其寡聚和激活。申请人实验室的初步证据表明，一种在自动炎性疾病家族性地中海发烧（FMF）中突变的蛋白质，通过与衔接蛋白ASC和procaspase-1组装炎症复合物，在炎症中起着重要作用，在炎症中起着重要作用。吡啶在凋亡中似乎也起着重要作用，因为与野生型小鼠的巨噬细胞相比，pyrin截断小鼠的巨噬细胞在凋亡中表现出A缺陷，响应于IL-4和LPS刺激。有证据表明，炎症性或可能的凋亡活性受两种被称为PSTPIP1和PSTPIP2的细胞骨架相关蛋白的调节。这些蛋白质中的错义突变与两种进一步的自动炎性疾病（称为化脓性关节炎，脓蛋白疾病，牙菌根综合征（PAPA）和慢性多灶性骨髓炎（CMO）综合征。在这种竞争连续应用中，提出了三个特定的目标，以研究吡啶在凋亡和炎症中的身体作用以及细胞骨架在这两个吡啶途径中的作用。在第一个具体目的中，将通过基于细胞和无细胞的重建系统以及人和小鼠巨噬细胞系中的详细生化和生物学研究来研究吡啶在caspase-1激活中的作用。在第二个特定目的中，提出了实验来表征PSTPIP1和PSTPIP2与吡啶的相互作用，并确定它们调节人和小鼠细胞中caspase-1激活的能力。在第三个特定目的中，生物学，生化和遗传方法将用于阐明吡啶调节细胞凋亡的机制。了解吡啶介导的炎症和凋亡反应的相互作用以及吡啶激活caspase-1的机制可能会为吡啶在先天免疫和凋亡中的作用提供重要的见解，并具有明显的潜在临床应用。公共卫生相关性陈述：编码吡啶和与吡啶相关的蛋白质PSTPIP1的基因中的突变与人类的严重自身炎症性疾病有关。这项研究将研究吡啶的身体功能，以阐明其在正常细胞功能中的作用以及与疾病相关的突变如何改变其功能。这项研究的结果将有助于设计和发现有效治疗自炎性疾病的疗法。