The role of ISG15 and ISGylation in the senescence-associated secretory phenotype

ISG15 和 ISGylation 在衰老相关分泌表型中的作用

• 批准号: 10788670
• 负责人: ALOYSIUS John KLINGELHUTZ
• 金额: $ 42.76万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10788670
• 关键词: Address; Affect; Aging; Brain; Cell Aging; Cell Line; Cell model; Cell secretion; Cells; DNA Damage; Data; Development; Disease; Dyskeratosis Congenita; Enzymes; Fibroblasts; Genes; Human; ISG15 gene; Infection; Inflammation; Interferon Inducers; Interferons; Kidney; Knock-in; Knock-out; Knockout Mice; Knowledge; Laboratories; Ligation; Liver; Malignant Neoplasms; Maps; Mediating; Methods; Modification; Mus; Muscle; Mutate; Mutation; Pathogenicity; Patients; Phenotype; Play; Premature aging syndrome; Proliferating; Proteins; Proteomics; Rattus; Rodent; Role; Sampling; Site; Skin; Stress; System; Technology; Telomerase; Telomere Shortening; Therapeutic Intervention; Tissues; Ubiquitin Like Proteins; Up-Regulation; Viral Oncogene; Virus Replication; age related; cell immortalization; cytokine; experimental study; genetic manipulation; human model; innovation; insight; mutant; pathogen; senescence; targeted treatment; telomere; tumor progression

Cellular senescence is a cellular state that affects age-related diseases such as inflammation, cancer development, and cancer progression. Cells that enter senescence have a senescence-associated secretory phenotype (SASP) that has major, and usually detrimental, effects on other cells in the surrounding microenvironment. ISG15 is a ubiquitin-like protein that can covalently modify host and pathogenic proteins, changing their stability and function. ISG15 can also be secreted and function as a cytokine. While ISG15 is classically regarded as an interferon-stimulated gene that is induced during infection by pathogens, it can also be upregulated by DNA damage and stress. Recent studies have demonstrated that ISG15 is among the most upregulated genes across tissues in aging rodents. It has also been shown by us and others that ISG15 is upregulated during replicative senescence of human fibroblasts. Our preliminary data further indicates that ISG15 is significantly upregulated in skin fibroblasts derived from patients with dyskeratosis congenita, a human model of dysfunctional telomerase and shortened telomeres. Immortalization of DC cells by telomerase activation restores ISG15 levels to normal. Little is known about how ISG15 upregulation contributes to senescence and what proteins are ISGylated in senescent cells. Further, it is unknown how ISGylation changes upon cell immortalization. We hypothesize that upregulation of ISG15 and concomitant ISGylation of proteins caused by DNA damage is important for the SASP. In this proposal, we will determine what the interplay is between secreted ISG15 and ISGylation in senescence and immortalization. Using our unique patient cells and cell lines, we will map the senescence-associated ISGylome using innovative proteomic strategies pioneered in our laboratories. We will compare the ISGylomes between cells that have been induced to senesce by different mechanisms and from human and mouse, and we will determine how secreted ISG15 affects surrounding cells. Further, we will assess how immortalization by telomerase or viral oncogenes changes the ISGylome. Finally, we will specifically determine how ISG15 and ISGylation affects the SASP through genetic manipulation of cells to have reduced and enhanced ISGylation. These studies will provide key insight into how upregulation of ISG15 and ISGylation of specific cellular proteins play a role in cellular senescence and, in particular, the SASP.

细胞衰老是一种细胞状态，影响与年龄有关的疾病，例如炎症，癌症 发育和癌症进展。进入衰老的细胞具有衰老相关分泌 表型（SASP）对周围其他细胞具有重大且通常有害的影响 微环境。 ISG15是一种泛素样蛋白，可以共价修改宿主和致病蛋白， 改变其稳定性和功能。 ISG15也可以分泌并用作细胞因子。而ISG15是 经典被视为由病原体在感染期间诱导的干扰素刺激的基因，也可以 被DNA损伤和压力上调。最近的研究表明，ISG15是最大的 衰老啮齿动物的组织上上调基因。我们和其他人也表明ISG15是 在人类成纤维细胞的复制衰老过程中上调。我们的初步数据进一步表明 ISG15在源自患有患者的患者的皮肤成纤维细胞中显着上调，它是人类 功能障碍端粒酶和端粒缩短的模型。通过端粒酶对DC细胞的永生化 激活将ISG15水平恢复为正常水平。关于ISG15上调的贡献知之甚少 衰老和哪种蛋白质在衰老细胞中被渗透。此外，未知iSgylation如何改变 细胞永生化。我们假设ISG15的上调和伴随的Isgylation 由DNA损伤引起的蛋白质对SASP很重要。在此提案中，我们将确定 相互作用在衰老和永生化中分泌的ISG15和Isgylation之间。使用我们独特的病人 细胞和细胞系，我们将使用创新的蛋白质组学策略来绘制与衰老相关的iSgylome 在我们的实验室中率先。我们将比较已诱导的细胞之间的iSgylomes 通过不同的机制以及人类和小鼠，我们将确定分泌的ISG15如何影响 周围的细胞。此外，我们将评估端粒酶或病毒癌基因的永生化如何改变 Isgylome。最后，我们将明确确定ISG15和Isgylation如何通过遗传影响SASP 操纵细胞以减少和增强的Isgylation。这些研究将提供有关如何 ISG15的上调和特定细胞蛋白的ISGyLation在细胞衰老中起作用，在 特别是SASP。