Role of O-GlcNAc-ylation on tumor progression

O-GlcNAc 基化在肿瘤进展中的作用

基本信息

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
项目类别：
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

项目摘要

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控制着关键的信号传导和生物过程，例如信号。 O-GlcNAc 稳态的扰动。葡萄糖水平升高与糖尿病、癌症和神经退行性疾病有关。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，包括我们最近报道，抑制 HBP 活性可显着降低乳腺癌的侵袭表型。我们推测，大量的葡萄糖（一种易于代谢的碳水化合物）会驱动乳腺肿瘤细胞的生长。通过 HBP 的通量，导致通过 O-GlcNAc 化修饰的蛋白质组合的富集。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- 重要的是，我们在 Hedgehog (Hh) 通路的转录因子中发现了修饰的 GLI 蛋白。血糖升高时，GLI 的 O-GlcNAc 修饰会恶化 Hh/GLI 活性并抑制 HBP；我们认为 HBP 引导的 O-GlcNAc 化从根本上来说是侵入性的，并且是有效的。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。确定 GLI O-GlcNAc 化的原因和后果我们将首先确定 HBP 的参与。接下来，我们将进行研究以确定 GLI 蛋白的 O-GlcNAc 修饰。 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为了丰富相关性，我们还将评估人类 TNBC 和 PDX 模型。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。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，进展和抗肿瘤药物的反应。