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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的基因组不稳定性是癌症和年龄相关疾病的主要驱动力。尽管长期以来，DNA 损伤反应被认为可以调节基因组完整性和细胞命运，越来越多的证据表明基因组不稳定性也会引发炎症反应。最近的研究深入了解了此类反应的机制证明胞质 DNA 传感器、环 GMP-AMP 合酶 (cGAS)，可以在 DNA 损伤与先天免疫的联系中发挥关键作用。有趣的是，我们的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来胞浆双链 DNA 做出反应，我们证明人类神经胶质细胞显示静止状态和激活后 cGAS 蛋白表达的稳定水平。此外，我们表明这些细胞类型组成型表达关键的下游 cGAS 接头蛋白，干扰素基因刺激剂 (STING)。在这项 R03 试点研究中，我们将开始检验 cGAS-STING 通路检测细胞质的假设基因毒性应激后小胶质细胞和/或星形胶质细胞中的 DNA 并启动神经胶质细胞自动炎症反应。该项目源于两家公司之间的新合作经验丰富的研究人员在 DNA 损伤研究方面具有互补的专业知识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 损伤是否会引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，我们将将这种反应与自身炎症介质的产生相关联或潜在的抗肿瘤因子，例如 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。这拟议的 R03 试点研究是这一新研究方向重要的第一步，将为更全面地调查该机构的作用提供坚实的理由 CNS 细胞衰老和癌症中的 cGAS-STING 通路，未来 R01 将寻求机制支持。