Immune control and genomic instability at micronuclei

微核的免疫控制和基因组不稳定性

• 批准号: 10544747
• 负责人: JOHN MACIEJOWSKI
• 金额: $ 49.28万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10544747
• 关键词: Anaphase; Automobile Driving; Biochemical; Cell Nucleus; Cell division; Cells; Cellular biology; Chromatin; Chromosomal Instability; Chromosome Segregation; Chromosomes; Chronic; Collaborations; Complex; Crows; Cytoplasm; DNA; DNA Binding; DNA Damage; DNA Sequence Rearrangement; DNA-Binding Proteins; Data; Defect; Digestion; Disease; Docking; Electron Transport Complex III; Endoplasmic Reticulum; Environment; Etiology; Evolution; Exhibits; Functional disorder; Gene Rearrangement; Genes; Genome; Genomic DNA; Genomic Instability; Goals; Health; Human; Immune; Immune System Diseases; Immune signaling; Incentives; Individual; Inflammatory; Interphase; Laboratories; Lead; Letters; Malignant Neoplasms; Membrane; Methods; Missense Mutation; Mitosis; Modeling; Molecular; Mutagenesis; Mutate; Mutation; N-terminal; Nuclear; Nuclear Envelope; Numerical Chromosomal Abnormality; Pathway interactions; Pattern; Penetration; Phase; Play; Positioning Attribute; Process; Prognosis; Proteins; Proteomics; Resected; Role; Rupture; Site; Source; Stimulator of Interferon Genes; Structural Chromosomal Abnormality; Structure; TREX1 gene; TREX2 gene; Testing; Therapeutic; Three Prime Repair Exonuclease 1; Topoisomerase; Tumor Immunity; Vesicle; Viral; Work; anti-tumor immune response; cancer genome; chromosome missegregation; chromothripsis; endonuclease; genome integrity; immune activation; immunoregulation; improved; insight; micronucleus; mutation carrier; novel; polyproline; prevent; protein protein interaction; pseudotoxoplasmosis syndrome; recruit; repaired; segregation; sensor; therapy resistant; tumor

PROJECT SUMMARY Chromosomal instability (CIN) is a hallmark of cancer characterized by high rates of chromosome mis-segre- gation during cell division. CIN can generate nuclear aberrations termed micronuclei when a chromosome or chromosome fragment lags during anaphase and fails to join the main chromatin mass that will form the prima- ry nucleus. Micronuclei recruit nuclear envelopes but defects in construction lead to frequent rupturing, loss of compartmentalization, and an unregulated exchange of proteins and small vesicles with the cytoplasm. Mi- cronuclear envelope rupturing causes broad dysfunction and is associated with extensive DNA damage and genomic rearrangements, including clustered mutational phenomena such as chromothripsis and kataegis, which are commonly observed in cancer genomes. Ruptured micronuclei can also activate the pro-inflammato- ry cGAS-STING pathway, which plays essential roles in anti-tumor immunity. These observations suggest that micronuclei may represent key platforms for genome evolution and immune activation in cancer. The mecha- nisms driving DNA damage and immune activation at micronuclei are poorly understood. The laboratory dis- covered that the endoplasmic reticulum (ER)-associated exonuclease TREX1, which is mutated in a variety of human immune diseases including Aicardi-Goutières Syndrome, accumulates at micronuclei upon micronu- clear envelope rupture where it resects micronuclear DNA and limits cGAS-STING activation. Therefore, TREX1 occupies central positions in key pathways with diverse roles in human health and disease. Conse- quently, there is strong rationale to understand mechanisms of TREX1 activity and engagement with cytosolic DNA. The long-term goals of the laboratory are to determine mechanisms of DNA damage, clustered mutage- nesis, and immune activation at sites of nuclear envelope rupture. The specific Aims of this proposal are to 1) Elucidate mechanisms of TREX1 structure and function, 2) Determine how TREX1 is recruited to micronuclei, and 3) Dissect pathways of micronuclear DNA damage. Each objective is supported by extensive preliminary data. Aim 1 will focus on a previously uncharacterized region in TREX1, which is essential for its ability to de- grade cytosolic DNA and inhibit cGAS activation. Aim 2 will build on results showing that TREX1 DNA binding function is dispensable for its localization to micronuclei, while its association with the ER is essential. Aim 3 will use a new method to purify micronuclei to dissect sources of micronuclear DNA damage. Taken together, these data will provide fundamental insights into cancer genome evolution, explain how previously uncharac- terized TREX1 mutations cause Aicardi-Goutières syndrome, and may identify new strategies to improve anti- tumor immunity.

项目摘要 染色体不稳定性（CIN）是癌症的标志，其特征是高染色体渗透疾病的率 细胞分裂期间的门。 CIN可以在染色体或 后期期间染色体片段滞后，未能连接主染色质质量，这将形成底漆 ry核。微核募集核信封，但建筑缺陷导致经常破裂，丧失 隔室化，以及蛋白质和小蔬菜与细胞质的不受调节的交换。 m 农作物包膜破裂会引起广泛的功能障碍，并与广泛的DNA损伤和 基因组重排，包括簇状的突变现象，例如铬胸和kataegis， 通常在癌症基因组中观察到。破裂的微核还可以激活pro-Inflammato- RY CGAS丁字道，在抗肿瘤免疫中起着重要作用。这些观察表明 微核可能代表了癌症中基因组进化和免疫激活的关键平台。机械 - 在微核驱动DNA损伤和免疫激活的NISM知之甚少。实验室 涵盖内质网（ER）相关的外切核酸外切酶Trex1，该核酸酶Trex1在多种 人类免疫疾病包括Aicardi-Goutières综合征，在微核时积聚在微核综合征 清除包膜破裂，它切除了微核DNA并限制CGAS刺激激活。所以， TREX1在人类健康和疾病中发挥不同作用的主要途径中占据中心位置。 con 有很强的理由来了解TREX1活动的机制和与胞质的互动 脱氧核糖核酸。实验室的长期目标是确定DNA损伤的机制，聚集的突变 - NES，核包膜破裂部位的免疫激活。该提议的具体目的是1） 阐明TREX1结构和功能的机制，2）确定如何将TREX1募集到微核， 3）剖析微核DNA损伤的途径。每个目标都得到广泛的初步支持 数据。 AIM 1将重点放在TREX1中以前未表征的区域上，这对于其具有脱颖而出的能力至关重要 等级的胞质DNA并抑制CGA激活。 AIM 2将基于结果表明TREX1 DNA结合 功能可用于将其定位到微核，而其与ER的关联至关重要。目标3 将使用一种新方法净化微核，以剖析微核DNA损伤的来源。在一起， 这些数据将提供对癌症基因组进化的基本见解，解释以前如何不符合 terex1突变引起aicardi-goutières综合征，并可能确定改善反抗的新策略 肿瘤免疫。