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 ß-N-乙酰氨基葡萄糖 (O-GlcNAc) 是附着在丝氨酸或丝氨酸侧链羟基上的糖。 蛋白质上的苏氨酸残基。 O-GlcNAcylation 控制关键信号传导和生物过程，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。葡萄糖水平增加会导致通量增加 通过己糖胺生物合成途径 (HBP)，最终导致 O-GlcNAc 水平增加。 HBP 的活性以及随后的细胞 O-GlcNAc 化在多种癌症类型中升高，包括 乳腺癌。我们最近报道，抑制 HBP 活性可显着降低侵袭表型 乳腺肿瘤细胞。我们推测，大量的葡萄糖（一种易于代谢的碳水化合物）会驱动 通过 HBP 的通量，导致通过 O-GlcNAc 化修饰的蛋白质组合的富集。使用 无偏见的蛋白质组学分析，我们发现升高的葡萄糖培养条件富含 O-GlcNAc- 修饰的 GLI 蛋白，Hedgehog (Hh) 通路的转录因子。重要的是，我们发现在 葡萄糖水平升高时，GLI 的 O-GlcNAc 修饰会加剧 Hh/GLI 活性；并抑制HBP 减轻了这种影响。我们假设 HBP 引导的 O-GlcNAc 化从根本上编程了侵入性和 通过激活 Hh/GLI 信号传导来增强肿瘤细胞的化学抗性。 在目标 1 中，我们将确定 HBP 指导的 GLI O-GlcNAc 化的分子基础。我们将 确定 GLI O-GlcNAc 化的原因和后果。我们将首先确定 HBP 的参与情况 GLI 蛋白的 O-GlcNAc 修饰。接下来，我们将进行调查以确定 HBP 信号如何参与 O-GlcNAc 修饰的 GLI 来编程侵入性和 肿瘤细胞的化学抗性。 在目标 2 中，我们将评估 O-GlcNAc 景观升高对分子和细胞属性的影响 使用两种不同且互补的同基因研究乳腺肿瘤和相关的免疫微气候 乳腺癌小鼠模型。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 活性是否可以消除 O-的影响 GlcNAc 基化对乳腺肿瘤细胞侵袭性和耐药性的影响。 相关性：我们提出的研究旨在系统地研究 O-GlcNAc 如何驱动代谢 癌细胞中的重编程在分子水平上连接，异常激活 Hh/GLI 信号传导。这 累积的结果将建立对 O-GlcNAc 化如何编程肿瘤侵袭的机制理解， 进展和对抗肿瘤的反应。