Role of O-GlcNAcome on Breast Cancer Initiating Cells

O-GlcNAcome 对乳腺癌起始细胞的作用

• 批准号: 10521975
• 负责人: Lauren Elizabeth Ball
• 金额: $ 6.99万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10521975
• 关键词: Breast Cancer Cell; Cancer Control; Cancer Model; Cell Maintenance; Cells; Drug Targeting; Drug resistance; Epithelial; Grant; Growth; In Vitro; Malignant Neoplasms; Mammospheres; Metabolic; Metabolic Pathway; Molecular; Natural regeneration; Neoplasm Metastasis; Nutrient; O-GlcNAc transferase; Pathway interactions; Pharmacology; Play; Proliferating; Property; Proteins; Radiation therapy; Regulation; Relapse; Resistance; Role; Signal Transduction; Testing; Therapeutic; Time; base; cancer cell; cancer initiation; cancer stem cell; cancer therapy; chemotherapy; conventional therapy; design; detection of nutrient; experimental study; glycosylation; in vivo; inhibitor; malignant breast neoplasm; new therapeutic target; novel; overexpression; pre-clinical; refractory cancer; self-renewal; sensor; stem cell biomarkers; stem-like cell; therapeutic target; therapeutically effective; tumor; tumor initiation

Project Summary It is widely accepted that tumors are highly heterogeneous. There is a subpopulation of cells in a tumor, called tumor-initiating cell, that can be isolated and are able to self-renew, differentiate and form the bulk of the tumor. Many cancers don’t respond to traditional chemotherapy or radiotherapy, and those that initially respond, often relapse. Conventional therapy only attacks proliferating cells, leaving behind a pool of resistant stem-like cells that are able to regenerate the whole tumor. Understanding mechanisms that regulate tumor-initiating activity will lead to designing and developing effective therapeutics. Our lab has demonstrated for the first time that the nutrient sensor O-GlcNAc transferase (OGT) regulates cancer-initiating cells in vitro and in vivo. Reducing OGT, genetically or pharmacologically, blocks mammosphere formation in vitro and reduced epithelial-mesechymal markers (EMT), cancer stem cell markers. Importantly, overexpression of OGT, in multiple breast cancer cells, increases cancer stem cell markers including NANOG, increases mammosphere formation in vitro and increases tumor initiation in vivo. In this proposal, we hope to uncover molecular mechanism by which OGT regulates tumor initiation, by in part, understanding OGT interactome and O-GlcNAcome in breast cancer tumor initiating cells. This information will allow us to identify novel therapeutic targets in treating cancer and reverse drug resistance. Based on our preliminary results, the central hypothesis of this application is that the nutrient sensor O- GlcNAc transferase plays a fundamental role in breast cancer initiating cells via, in part, NANOG regulation. Completion of these experiments will contribute to our understanding of how nutrient-sensing pathways connects at the molecular level to self-renewing cancer stem cells (CSCs) and providing a framework for understanding how cancer alterations in metabolic pathways regulate core self-renewal signaling that controls CSC maintenance. In Aim #1, we will determine the molecular basis of OGT/O- GlcNAc regulation of the master CSC regulator NANOG. This aim will determine the molecular basis of OGT regulation of NANOG in breast cancer tumor initiating cells.In Aim #2, we will Identify OGT interactome/O- GlcNAcome between between cancer cells and cancer stem cells. This aim will identify OGT interacting proteins and O-GlcNAcylated protein in tumor initiating cells to identify novel pathways and regulators of tumor-initiating ability. The final aim will evaluate the role of OGT in regulating tumor-initiating activity in vivo. Importantly, we will test novel OGT inhibitors in preclinical cancer models and test whether OGT targeting drugs as potential anti-tumor initiation cell therapeutic strategy against breast cancer growth and metastasis in vivo. These studies will further our understanding of how metabolic reprogramming in cancer cells connects at the molecular level to tumor initiating cells and will create mechanistic understanding of how nutrient sensor OGT can couple to cancer initiation pathways and establish OGT as therapeutic target for treatment of resistant cancers.

项目摘要 人们普遍认为肿瘤是高度异质性的。肿瘤中有一个细胞亚群，称为 肿瘤起始细胞，其可以被分离并且能够自我更新、分化和形成肿瘤的主体。 许多癌症对传统的化疗或放疗没有反应，而那些最初 反应，经常复发。传统疗法只攻击增殖细胞，留下一个耐药细胞池。 能够再生整个肿瘤的干细胞样细胞。了解调控机制 肿瘤引发活性将导致设计和开发有效的治疗剂。我们的实验室已经证明 营养传感器O-GlcNAc转移酶（OGT）首次在体外调节癌症起始细胞 和体内。减少OGT，遗传或非遗传，在体外阻断乳腺球形成 以及减少的上皮-间质标记物（EMT）、癌症干细胞标记物。重要的是， 在多种乳腺癌细胞中，OGT的过表达增加了癌症干细胞标志物，包括 NANOG在体外增加乳腺球形成并在体内增加肿瘤起始。在这 因此，我们希望通过对OGT的研究，揭示OGT调控肿瘤发生的分子机制， 第二部分，了解OGT相互作用组和O-GlcNAcome在乳腺癌肿瘤起始细胞中的作用。这 这些信息将使我们能够确定治疗癌症和逆转耐药性的新治疗靶点。 基于我们的初步结果，该应用的中心假设是营养传感器O- GlcNAc转移酶在乳腺癌起始细胞中起着重要作用，部分通过NANOG 调控这些实验的完成将有助于我们理解营养感应是如何 在分子水平上与自我更新的癌症干细胞（CSCs）连接，并提供了一个 理解癌症代谢途径改变如何调节核心自我更新的框架 控制CSC维护的信令。在目标#1中，我们将确定OGT/O-的分子基础。 主CSC调节剂NANOG的GlcNAc调节。这一目标将确定OGT的分子基础 在目标#2中，我们将鉴定OGT相互作用组/O- GlcNA介于癌细胞和癌干细胞之间。这一目标将确定OGT相互作用 蛋白质和O-GlcNAc酰化蛋白质在肿瘤起始细胞中鉴定新的途径和调节剂 引发肿瘤的能力。最终的目的是评价OGT在调节肿瘤启动过程中的作用。 体内活性。重要的是，我们将在临床前癌症模型中测试新型OGT抑制剂， OGT靶向药物是否可作为潜在的抗乳腺癌起始细胞治疗策略 体内癌症生长和转移。这些研究将进一步加深我们对代谢是如何 癌细胞中的重编程在分子水平上与肿瘤起始细胞连接， 对营养传感器OGT如何与癌症起始途径偶联的机制理解， 确立OGT作为治疗抗性癌症治疗靶点。