Decoding protein MARylation networks in astrocytes using chemical biology approaches

使用化学生物学方法解码星形胶质细胞中的蛋白质 MARylation 网络

• 批准号: 10599222
• 负责人: Michael S Cohen
• 金额: $ 50.3万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10599222
• 关键词: ADP ribosylation; Address; Adenosine Diphosphate Ribose; Alzheimer' s Disease; Amino Acids; Area; Astrocytes; Bacteria; Binding; Biology; Brain; Brain Injuries; Cell Membrane Permeability; Cell physiology; Cells; Cellular biology; Central Nervous System; Chemicals; Chemistry; Collaborations; Data; Development; Endowment; Engineering; Environment; Enzymes; Family; Family member; Funding; Generations; Goals; Growth Factor; Health; Immune response; Immune signaling; Immunooncology; Individual; Innate Immune Response; Ischemic Stroke; Knowledge; Laboratories; Lead; Ligands; Literature; Mammalian Cell; Mammals; Mass Spectrum Analysis; Mediating; Mediator; Membrane; Multiple Sclerosis; Natural Immunity; Nature; Nerve Degeneration; Nervous System Trauma; Neurodegenerative Disorders; Neurosciences; Nicotinamide adenine dinucleotide; Nucleotides; Pathogenesis; Pathology; Pathway interactions; Phenotype; Physiological; Poly(ADP-ribose) Polymerase Inhibitor; Post-Translational Protein Processing; Process; Protein Engineering; Proteins; Role; Shapes; Signal Transduction; Site; Stimulus; Structure; Testing; Toxin; Variant; Virus; Work; acute infection; analog; astrogliosis; cell type; chemical synthesis; chronic inflammatory disease; design; high risk; immunoregulation; improved; inhibitor; insight; knockout gene; member; multidisciplinary; neuroinflammation; novel; novel therapeutic intervention; pathogen; response; spatiotemporal; steroid hormone; tandem mass spectrometry; tool

Project Summary Astrocytes are critical regulators of innate immunity in the central nervous system (CNS). Stimulation of CNS innate immunity by neuroinflammatory activators such as pathogens and brain injury, as well as in response to neurodegeneration, cause astrocytes to undergo a transition to a reactive phenotype called astrogliosis. While it is well accepted that astrogliosis can act as a protective mechanism to minimize CNS damage, the mechanisms that regulate astrogliosis are not well understood. Our preliminary results and data from the literature support our general hypothesis that PARP7 controlled MARylation critically shapes the innate immune responses in the CNS. Our long-term goal is to understand the role of PARP7 in astrogliosis and whether PARP7 represents an actionable target for CNS pathologies that arise as a consequence of activation of CNS innate immunity. The objective of the proposed work is elucidate the mechanisms by which PARP7 regulates innate immunity in astrocytes. PARP7 has emerged as a critically important member of a large enzyme family known as PARPs, especially in the innate immune response. Similar to other PARP family members, PARP7 catalyzes the post- translational modification known as mono-ADP-ribosylation (MARylation), which involves the transfer of ADP-ribose from NAD+ to amino acids on target proteins. The MARylation targets of PARP7 in astrocytes are unknown. To decode the mechanisms by which PARP7 regulates innate immunity in astrocyte, we need to identify the direct targets of PARP7 in astrocytes. Identifying the direct targets of PARP7 has been challenging, however, due to the fact that PARPs share the same substrate NAD+. To overcome this limitation, we describe the development of engineered PARP7—orthogonal NAD+ analogue pairs for identifying the direct targets of PARP7 in astrocytes lysates (Aim I). We also describe the generation of membrane-permeant variants of our orthogonal NAD+ analogues, which are critical for identifying PARP7 targets in intact astrocytes using stimuli that activate the innate immune response in astrocytes (Aim II). Lastly, we describe a strategy for improving the selectivity of PARP7 inhibitors (Aim III). Selective inhibitors of PARP7 are essential chemical probes for evaluating the function of PARP7-mediated MARylation in the innate immune response in astrocytes. We anticipate that these studies will not only clarify our understanding of the function of PARP7-mediated MARylation in innate immunity in astrocytes, but could also lead to new therapeutic strategies for CNS pathologies, particularly neuroinflammatory (e.g. multiple sclerosis) and neurodegenerative diseases (e.g. Alzheimer's disease). More generally, the results obtained from these studies will have far-reaching impact on our understanding of MARylation in cell signaling.

项目摘要 星形胶质细胞是中枢神经系统（CNS）先天免疫的关键调节剂。刺激中枢神经系统 神经炎症激活剂（例如病原体和脑损伤）的先天免疫力，以及对 神经变性，导致星形胶质细胞过渡到称为星形胶质细胞增多的反应性表型。同时 被广泛接受的是，星形胶质症可以充当最大程度地减少中枢神经系统损伤的保护机制 调节星形胶质细胞增多尚未得到很好的了解。我们的初步结果和文献支持的数据 我们的总体假设PARP7控制了玛丽化，批判性地塑造了先天的免疫反应 中枢神经系统。我们的长期目标是了解PARP7在星形胶质细胞增多症中的作用以及PARP7是否代表 CNS先天免疫激活而产生的CNS病理的可行靶标。这 拟议工作的目的是阐明PARP7调节先天免疫力的机制 星形胶质细胞。 PARP7已成为一个非常重要的大型酶家族的重要成员，称为PARPS， 特别是在先天免疫反应中。与其他PARP家族成员类似，PARP7催化了后 转化修饰称为单ADP-核糖基化（MARYLATION），涉及转移 靶蛋白上的NAD+到氨基酸的ADP-核糖。星形胶质细胞中PARP7的玛丽化目标是 未知。为了解码PARP7调节星形胶质细胞先天免疫的机制，我们需要 确定星形胶质细胞中PARP7的直接靶标。确定PARP7的直接目标一直是挑战， 但是，由于PARP共享相同的基板NAD+。为了克服这一限制，我们描述 工程PARP7的开发 - 正交NAD+模拟对，用于识别直接目标 星形胶质细胞中的PARP7（AIM I）。我们还描述了我们 正交NAD+类似物，这对于使用Stimulli识别完整星形胶质细胞中的PARP7靶标至关重要 激活星形胶质细胞中的先天免疫反应（AIM II）。最后，我们描述了改进的策略 PARP7抑制剂的选择性（AIM III）。 PARP7的选择性抑制剂是必需的化学问题 评估星形胶质细胞中先天免疫反应中PARP7介导的玛丽化的功能。我们 预计这些研究不仅会阐明我们对PARP7介导的功能的理解 玛丽特在星形胶质细胞中具有先天免疫力，但也可能导致CNS的新治疗策略 病理，特别是神经炎症性（例如多发性硬化症）和神经退行性疾病（例如， 阿尔茨海默氏病）。更一般地，从这些研究中获得的结果将对我们的我们产生深远的影响 了解细胞信号中的玛丽化。