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: 10378049
• 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/10378049
• 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; Chemicals; Chemistry; Collaborations; Data; Development; 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; Masks; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Membrane; Multiple Sclerosis; Natural Immunity; Nature; Nerve Degeneration; Nervous System Trauma; Neuraxis; Neurodegenerative Disorders; Neurosciences; Nicotinamide adenine dinucleotide; Nucleotides; Pathogenesis; Pathology; Pathway interactions; Phenotype; Physiological; 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.

项目概要 星形胶质细胞是中枢神经系统（CNS）先天免疫的关键调节因子。刺激中枢神经系统 病原体和脑损伤等神经炎症激活剂的先天免疫，以及对 神经变性，导致星形胶质细胞转变为称为星形胶质细胞增生的反应性表型。虽然它 人们普遍认为，星形胶质细胞增生可以作为一种保护机制，最大限度地减少中枢神经系统损伤，该机制 调节星形胶质细胞增生的机制尚不清楚。我们的初步结果和文献数据支持 我们的一般假设是 PARP7 控制的 MARylation 关键地塑造了先天免疫反应 中枢神经系统。我们的长期目标是了解 PARP7 在星形胶质细胞增生中的作用以及 PARP7 是否代表 由于中枢神经系统先天免疫激活而产生的中枢神经系统病理的可操作目标。这 拟议工作的目标是阐明 PARP7 调节先天免疫的机制 星形胶质细胞。 PARP7 已成为 PARPs 大酶家族中极其重要的成员， 尤其是在先天免疫反应中。与其他 PARP 家族成员类似，PARP7 催化后 翻译修饰称为单 ADP 核糖基化 (MARylation)，其中涉及转移 ADP-核糖从 NAD+ 转化为靶蛋白上的氨基酸。星形胶质细胞中 PARP7 的 MARylation 靶点是 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 的直接目标一直具有挑战性， 然而，由于 PARP 共享相同的底物 NAD+。为了克服这个限制，我们描述 工程化 PARP7 的开发——正交 NAD+ 类似物对，用于识别直接靶标 星形胶质细胞裂解物中的 PARP7（目标 I）。我们还描述了我们的膜渗透变体的产生 正交 NAD+ 类似物，对于使用刺激识别完整星形胶质细胞中的 PARP7 靶标至关重要 激活星形胶质细胞的先天免疫反应（目标 II）。最后，我们描述了改进策略 PARP7 抑制剂的选择性（目标 III）。 PARP7 的选择性抑制剂是重要的化学探针 评估 PARP7 介导的 MARylation 在星形胶质细胞先天免疫反应中的功能。我们 预计这些研究不仅会澄清我们对 PARP7 介导的功能的理解 星形胶质细胞先天免疫中的 MARylation 也可能导致中枢神经系统的新治疗策略 病理学，特别是神经炎症（例如多发性硬化症）和神经退行性疾病（例如神经退行性疾病） 阿尔茨海默病）。更一般地说，这些研究获得的结果将对我们产生深远的影响。 了解细胞信号转导中的 MARylation。