cGAS-mediated glial responses to DNA damage: A pilot study

cGAS 介导的神经胶质细胞对 DNA 损伤的反应：一项初步研究

• 批准号: 9893482
• 负责人: Christine A. Richardson
• 金额: $ 7.14万
• 依托单位: UNIVERSITY OF NORTH CAROLINA CHARLOTTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893482
• 关键词: Adaptor Signaling Protein; Antitumor Response; Appearance; Astrocytes; Binding; CRISPR/Cas technology; Cell Aging; Cell Nucleus; Cells; Central Nervous System Diseases; Collaborations; Cyclic GMP; Cytosol; DNA; DNA Damage; DNA Repair; Data; Development; Disease; Event; Future; Genetic Transcription; Genomic DNA; Genomic Instability; Genotoxic Stress; Goals; Grant; Human; Immune; Immune response; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Interferon Type I; Interferon-beta; Interferons; Interleukin-6; Investigation; Knock-out; Link; Malignant Neoplasms; Mediating; Mediator of activation protein; Messenger RNA; Microglia; Mitochondria; Molecular; Mus; NF-kappa B; Natural Immunity; Neuraxis; Neurodegenerative Disorders; Neuroglia; Nuclear; Nucleic Acids; Pathway interactions; Pattern; Pharmacology; Pilot Projects; Play; Production; Research; Research Personnel; Research Project Grants; Rest; Roentgen Rays; Role; Rupture; Solid; Stimulator of Interferon Genes; TNF gene; Techniques; Testing; age related; autoinflammatory; brain cell; cell type; cytokine; driving force; ds-DNA; experience; genome editing; genome integrity; immune function; inhibitor/antagonist; insight; irradiation; novel; protein expression; response; senescence; sensor; small molecule; stem; tumor

Abstract Genomic instability is a major driving force for cancer and age-related diseases. While DNA damage responses were long thought to regulate genome integrity and cell fates, evidence is accumulating that genomic instability also triggers inflammatory responses. Recent studies have provided insight into the mechanisms underlying such responses with the demonstration that the cytosolic DNA sensor, cyclic GMP-AMP synthase (cGAS), can play a key role in linking DNA damage to innate immunity. Interestingly, our research team has recently described the ability of human microglia and astrocytes to respond to foreign cytosolic double-stranded DNA and we demonstrated that human glia show robust levels of cGAS protein expression at rest and following activation. Furthermore, we showed these cell types constitutively express the critical downstream cGAS adaptor protein, stimulator of interferon genes (STING). In this R03 pilot study, we will begin to test the hypothesis that the cGAS-STING pathway detects cytoplasmic DNA in microglia and/or astrocytes after genotoxic stress and initiates glial auto- inflammatory responses. This project stems from a new collaboration between two experienced investigators with complementary expertise in the study of DNA damage repair mechanisms and glial innate immune sensor molecules. In these preliminary studies, we will determine whether genomic DNA damage elicits micronuclei formation and an elevation in the level of the cGAS product cGAMP in cultured glia, and we will correlate such responses with the production of auto-inflammatory mediators or potentially anti-tumor factors such as type I interferon. Furthermore, we will directly assess the relative importance of the cGAS-STING pathway in glial responses to DNA damage following pharmacological inhibition and/or CRISPR/cas9 genome editing. The proposed pilot R03 studies are an important first step in this new research direction and will provide a solid rationale for a more comprehensive investigation into the role of the cGAS-STING pathway in CNS cellular senescence and cancer for which future R01 mechanism support will be sought.

摘要 基因组不稳定性是癌症和与年龄有关的疾病的主要驱动力。而 DNA损伤反应长期以来被认为是调节基因组完整性和细胞命运的， 越来越多的证据表明，基因组的不稳定性也会引发炎症反应。 最近的研究提供了对这种反应背后的机制的深入了解 证明了细胞质DNA传感器，环GMP-AMP合酶 cGAS（cGAS）可以在将DNA损伤与先天免疫联系起来方面发挥关键作用。有趣的是，我们 一个研究小组最近描述了人类小胶质细胞和星形胶质细胞的能力， 对外源胞质双链DNA的反应，我们证明了人类神经胶质细胞 在静止和活化后显示出强水平的cGAS蛋白表达。 此外，我们发现这些细胞类型组成性表达关键的下游蛋白。 cGAS衔接蛋白，干扰素基因刺激因子（STING）。在R 03初步研究中， 我们将开始检验cGAS-STING途径检测细胞质内 基因毒性应激后小胶质细胞和/或星形胶质细胞中的DNA，并启动胶质细胞自 炎症反应。这个项目源于两个人之间的新合作 在DNA损伤研究方面具有互补专业知识的经验丰富的研究人员 修复机制和神经胶质先天免疫传感器分子。在这些初步 研究中，我们将确定基因组DNA损伤是否诱发微核形成 以及培养的神经胶质细胞中cGAS产物cGAMP水平的升高，我们将 将这些反应与自身炎症介质的产生相关联，或 潜在的抗肿瘤因子如I型干扰素。此外，我们将直接 评估cGAS-STING通路在神经胶质细胞对DNA反应中的相对重要性 药理学抑制和/或CRISPR/cas9基因组编辑后的损伤。的 拟议的R 03试点研究是这一新研究方向的重要第一步， 将提供一个坚实的理由，更全面的调查的作用， cGAS-STING通路在CNS细胞衰老和癌症中的作用，未来R 01 将寻求机制支持。