Role of O-GlcNAc-ylation on tumor progression

O-GlcNAc 基化在肿瘤进展中的作用

• 批准号: 10589810
• 负责人: Lalita A. Shevde
• 金额: $ 33.29万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-10 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10589810
• 关键词: ABCB1 gene; Antineoplastic Agents; Attenuated; Biological Models; Biological Process; Breast Cancer Cell; Breast Cancer Model; Carbohydrates; Cell Cycle Progression; Cell Proliferation; Cells; Characteristics; Chemicals; Chemoresistance; DNA Repair; DNA Repair Gene; Diabetes Mellitus; Diet; Embryonic Development; Erinaceidae; Event; GLI Family Protein; GLI gene; Genetic; Genetic Transcription; Glucose; Homeostasis; Human; Hydroxyl Radical; Hyperglycemia; Immune; Impairment; Investigation; Licensing; Link; Malignant Neoplasms; Mammary Neoplasms; Mass Spectrum Analysis; Metabolic; Metabolism; Microclimate; Modeling; Modification; Molecular; Neurodegenerative Disorders; Normal tissue morphology; Nutritional status; Outcome; Pathway interactions; Phenotype; Process; Proteins; Proteomics; Reporting; Research Design; Role; Serine; Side; Signal Pathway; Signal Transduction; Site; Structure; Testing; Threonine; Tissues; Tumor Cell Invasion; Up-Regulation; cancer cell; cancer type; chemotherapeutic agent; cytotoxic; glucose uptake; malignant breast neoplasm; mouse model; neoplastic cell; novel; patient derived xenograft model; programs; resilience; response; small molecule inhibitor; smoothened signaling pathway; success; sugar; tool; transcription factor; triple-negative invasive breast carcinoma; tumor; tumor progression

Abstract O-linked ß-N-acetylglucosamine (O-GlcNAc) is a sugar attachment to the side chain hydroxyl of a serine or threonine residue on proteins. O-GlcNAcylation controls key signaling and biological processes such as signal transduction, transcription, cell cycle progression, and metabolism. Perturbations in O-GlcNAc homeostasis have been linked with diabetes, cancer, and neurodegenerative diseases. Increased glucose levels channel flux through the Hexoseamine Biosynthetic Pathway (HBP), culminating in increased O-GlcNAc levels. The activity of HBP and consequently cellular O-GlcNAc-ylation are elevated in several cancer types, including breast cancer. We recently reported that inhibiting HBP activity significantly decreased the invasive phenotype of breast tumor cells. We surmised that abundance of glucose, a readily-metabolizable carbohydrate, will drive flux through HBP, resulting in enrichment of a portfolio of proteins that are modified by O-GlcNAc-ylation. Using unbiased proteomics analysis, we identified that elevated glucose culture conditions enrich for O-GlcNAc- modified GLI proteins, transcription factors of the Hedgehog (Hh) pathway. Importantly, we identified that in elevated glucose conditions, O-GlcNAc-modification of GLI exacerbates Hh/GLI activity; and inhibiting HBP mitigated this effect. We hypothesize that HBP-directed O-GlcNAc-ylation fundamentally programs invasive and chemoresistant attributes in tumor cells through activating Hh/GLI signaling. In Aim 1 we will determine the molecular underpinnings of HBP-directed O-GlcNAc-ylation of GLI. We will determine the causes and consequences of GLI O-GlcNAc-ylation. We will first identify engagement of the HBP in O-GlcNAc-modification of GLI proteins. Next, we will undertake investigations to identify establish the mechanistic basis of how HBP signaling engages O-GlcNAc-modified GLI to program invasive and chemoresistant attributes in tumor cells. In Aim 2 we will evaluate the impact of an elevated O-GlcNAc landscape on molecular and cellular attributes of the mammary tumor and the associated immune microclimate using two distinct and complementary syngeneic mouse models of mammary cancer. To enrich the relevance, we will also evaluate human TNBC and PDX model systems. We will test if inhibiting GLI activity, in the context of elevated O-GlcNAc, uncouples the influence of O- GlcNAc-ylation on invasive and chemoresistant attributes of mammary tumor cells. Relevance: Our proposed studies are structured to systematically investigate how O-GlcNAc-driven metabolic reprogramming in cancer cells connects at the molecular level to aberrantly activate Hh/GLI signaling. The cumulative outcomes will create mechanistic understanding of how O-GlcNAc-ylation programs tumor invasion, progression and response to anti-neoplastics.

抽象的 O连接的ß-N-乙酰葡萄糖胺（O-GLCNAC）是糖的糖附着，或 蛋白质上的苏氨酸住所。 O-Glcnacylation控制关键信号传导和生物学过程，例如信号 转导，转录，细胞周期进程和代谢。 O-GLCNAC稳态的扰动 与糖尿病，癌症和神经退行性疾病有关。增加葡萄糖水平通道通量 通过六氨酰胺生物合成途径（HBP），达到O-GLCNAC水平的提高。 在几种癌症类型中，HBP的活性以及因此 乳腺癌。我们最近报道，抑制HBP活性显着降低了侵入性表型 乳腺肿瘤细胞。我们浏览了葡萄糖的抽象，一种易于超代谢的碳氢化物将驱动 通过HBP的通量，导致通过O-GlCNAC基化修饰的蛋白质组合富集。使用 公正的蛋白质组学分析，我们确定升高的葡萄糖培养条件富含O-GlcNAC- 修饰的Gli蛋白，刺猬（HH）途径的转录因子。重要的是，我们确定了 GLI加剧HH/GLI活性的葡萄糖条件升高，O-GlCNAC调节；并抑制HBP 减轻了这种效果。我们假设HBP指导的O-GLCNAC基因从根本上进行侵入性和 通过激活HH/GLI信号传导，肿瘤细胞中的化学抗性属性。 在AIM 1中，我们将确定GLI的HBP指导O-GlCNAC基化的分子基础。我们将 确定Gli O-GlcNAC基化的原因和后果。我们将首先确定HBP的参与 在Gli蛋白的O-GLCNAC修饰中。接下来，我们将进行调查，以确定建立 HBP信号传导如何使O-GLCNAC修饰的GLI与编程侵入性和 肿瘤细胞中的化学抗性。 在AIM 2中，我们将评估升高的O-GlCNAC景观对分子和细胞属性的影响 乳腺肿瘤和相关的免疫小气候使用两个不同的互补合成性 乳腺癌的小鼠模型。为了丰富相关性，我们还将评估人类TNBC和PDX模型 系统。我们将测试在O-GlcNAC升高的情况下，抑制GLI活性是否会抑制O-的影响。 GlcNAC基因在乳腺肿瘤细胞的浸润性和化学抗性属性上。 相关性：我们提出的研究的结构是系统地研究O-GLCNAC驱动的代谢 在癌细胞中重新编程在分子水平上连接，以异常激活HH/GLI信号传导。这 累积结果将使O-GlCNAC基因侵袭O-GlCNAC基因侵入的机械理解， 对抗塑料的进展和反应。