Regulation of necrotic cell death by protein kinase A and cylindromatosis

蛋白激酶 A 和圆柱瘤病对坏死细胞死亡的调节

基本信息

批准号：
7875932
负责人：
FRANCIS Kaming CHAN
金额：
$ 20.54万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-25 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7875932
关键词：
Address Adjuvant Affect Apoptosis Apoptotic Biological Assay Caspase Catalytic Domain Cell Death Cells Cessation of life Complement Complex Cyclic AMP-Dependent Protein Kinases Deubiquitination Development Disease Event Family member Genes Host Defense Immune response Immunity In Vitro Infectious Agent Inflammation Inflammatory Knowledge Malignant Neoplasms Mediating Modeling Molecular Necrosis Paper Pathway interactions Phosphorylation Phosphotransferases Physiological Play Polyubiquitination Process Protein-Serine-Threonine Kinases Proteins RIPK3 gene RNA Interference Recombinants Regulation Role Serine Signal Transduction Testing Threonine Tissues Tumor Suppressor Proteins Vaccinia virus Viral Virus Virus Diseases Virus Inhibitors cell injury cytokine human RIPK1 protein mutant programs public health relevance research study response

项目摘要

DESCRIPTION (provided by applicant): Programmed necrosis is a morphologically and molecularly distinct form of cell death from apoptosis. Cell death by necrosis releases endogenous adjuvants or "danger signals" into the tissue milieu, which can trigger inflammation and stimulate immune responses. A role for programmed necrosis in anti-viral immunity is supported by recent discoveries of viral inhibitors that block this pathway of cell death. Mechanistically, programmed necrosis requires the serine/threonine protein kinas RIP1 and is optimally induced when caspases are inhibited. These results highlight a possible role for programmed necrosis in host defense against infectious agents. In order to further understand the molecular mechanism that regulates programmed necrosis, we screened by small interference RNA (siRNA) kinases and cancer-related genes that may participate in this non-apoptotic cell death pathway. We recently described the critical role of another RIP family member identified from the screen, RIP3, in TNF-induced and virus-induced programmed necrosis. In addition to RIP3, protein kinase A (PKA) catalytic subunit b and the tumor suppressor cylindromatosis (CYLD) were also identified from the RNAi screen as crucial for programmed necrosis. In this application, we will examine the molecular mechanisms by which PKA-C2 and CYLD regulates RIP1/RIP3-dependent programmed necrosis. We will examine their recruitment to the pro-necrotic signaling complex and how they might regulate the formation of the pro-necrotic signaling complex. We will determine whether deubiquitinase activity of CYLD is required for its function and explore the possible substrates for CYLD during programmed necrosis. Finally, we will test the physiological relevance of PKA-C2 and CYLD in virus infections using in vitro vaccinia virus infection as a model. PUBLIC HEALTH RELEVANCE: Cell death by necrosis causes inflammation and can greatly impact the quality of an immune response. However, the signals that regulate this process are poorly understood. We have recently identified two molecules, PKA-C2 and CYLD, to be essential for necrotic cell death. In this proposal, we will elucidate the mechanisms by which these two molecules control cell death by necrosis. Furthermore, we will evaluate how these two molecules may affect the cell death response during virus infections. These studies will allow us to better understand the signals that control cell death and inflammation. Eventually, the knowledge gained from these studies will aid the development of strategies to control inflammatory diseases caused by cell injury.

描述（由申请人提供）：程序性坏死是一种在形态学和分子学上与细胞凋亡不同的细胞死亡形式。细胞坏死死亡会将内源性佐剂或“危险信号”释放到组织环境中，从而引发炎症并刺激免疫反应。最近发现的阻断细胞死亡途径的病毒抑制剂支持了程序性坏死在抗病毒免疫中的作用。从机制上讲，程序性坏死需要丝氨酸/苏氨酸蛋白激酶 RIP1，并且当半胱天冬酶受到抑制时，最能诱导程序性坏死。这些结果强调了程序性坏死在宿主防御感染因子中可能发挥的作用。为了进一步了解调节程序性坏死的分子机制，我们通过小干扰RNA（siRNA）激酶和可能参与这种非凋亡细胞死亡途径的癌症相关基因进行筛选。我们最近描述了从筛选中鉴定出的另一个 RIP 家族成员 RIP3 在 TNF 诱导和病毒诱导的程序性坏死中的关键作用。除了 RIP3 之外，RNAi 筛选还鉴定出蛋白激酶 A (PKA) 催化亚基 b 和肿瘤抑制柱状瘤病 (CYLD) 对程序性坏死至关重要。在本申请中，我们将研究 PKA-C2 和 CYLD 调节 RIP1/RIP3 依赖性程序性坏死的分子机制。我们将检查它们对促坏死信号复合物的募集以及它们如何调节促坏死信号复合物的形成。我们将确定 CYLD 的去泛素酶活性是否是其功能所必需的，并探索 CYLD 在程序性坏死过程中可能的底物。最后，我们将以体外牛痘病毒感染为模型，测试 PKA-C2 和 CYLD 在病毒感染中的生理相关性。公共卫生相关性：细胞坏死导致炎症，并可极大地影响免疫反应的质量。然而，人们对调节这一过程的信号知之甚少。我们最近发现了两种分子：PKA-C2 和 CYLD，它们对于坏死细胞死亡至关重要。在本提案中，我们将阐明这两种分子通过坏死控制细胞死亡的机制。此外，我们将评估这两种分子如何影响病毒感染期间的细胞死亡反应。这些研究将使我们能够更好地了解控制细胞死亡和炎症的信号。最终，从这些研究中获得的知识将有助于制定控制由细胞损伤引起的炎症性疾病的策略。