Nutrient Sensing and Transcriptional Regulation

营养感应和转录调节

基本信息

批准号：
10421294
负责人：
Donald E Ayer
金额：
$ 38.41万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10421294
关键词：
Affect Age Biology Biotin Branched-Chain Amino Acids Breast Cancer Cell Breast Cancer cell line Cancer Biology Catabolism Cell Culture Techniques Cells ChIP-seq Choices and Control Clinical Critical Pathways Dependence Disease Equilibrium Gene Expression Gene Expression Regulation Genes Genetic Transcription Genomic approach Glucose Glycolysis Goals Growth Hypoxia In Vitro Ketones Label Maintenance Malignant Neoplasms Metabolic Metabolism Mitochondria Molecular Normal Cell Nutrient Oncogenes Outcome Oxidative Phosphorylation Oxygen Pathologic Pathway interactions Play Proteins Role Shapes Signal Transduction Survival Rate TXNIP gene Testing Therapeutic Therapeutic Intervention Transcriptional Regulation Warburg Effect Xenograft procedure aerobic glycolysis aggressive breast cancer c-myc Genes cancer cell cancer subtypes cofactor detection of nutrient experimental study extracellular glucose uptake in vitro activity in vivo interest malignant breast neoplasm member neoplastic cell new therapeutic target novel oxidation patient derived xenograft model programs response sensor targeted treatment therapeutic development tissue culture transcription factor transcriptome transcriptome sequencing triple-negative invasive breast carcinoma tumor tumor growth tumor microenvironment tumor xenograft tumorigenesis

项目摘要

SUMMARY Breast cancer is a heterogeneous disease consisting of several clinically and molecularly distinct subtypes. Compared to other subtypes, Triple Negative Breast Cancer (TNBC), which represents 15-20% of all breast cancer, occurs at a younger age and recurs more frequently with a reduced survival rate. Currently, no targeted therapies are available for TNBC. Our short-term goal is to dissect the molecular pathways that support TNBC growth and survival. Our long-term goal is to leverage these discoveries to identify novel targets suitable for therapeutic development. We focus on how normal cells and cancer cells integrate intra- and extracellular growth and nutrient signals to sustain growth and tumorigenesis. We are most interested in how two members of the extended Myc network of transcription factors integrate growth and nutrient signals. c-Myc and MondoA function in opposition to control the aggressive growth and survival of TNBC. In this application we focus on the control of gene expression by MondoA and how metabolism shapes the MondoA-dependent transcriptome in vitro and in vivo. MondoA is the principal regulator of glucose-dependent transcription with Thioredoxin Interacting Protein (TXNIP) being its best-characterized direct and glucose-induced transcriptional target. TXNIP has pleiotropic function. Best characterized among these many functions is one in fuel choice. TXNIP is a potent suppressor of glucose uptake and aerobic glycolysis, but it can also drive the catabolism of other fuels such as branched chain amino acids and ketones in mitochondria. Thus, high TXNIP levels drive fuel oxidation in mitochondria, while restricting utilization of glucose. Conversely, low TXNIP levels tip fuel use towards glucose and away from mitochondria. We have shown that several predominant oncogenes block MondoA transcriptional activity, thereby decreasing TXNIP levels. This likely contributes to oncogene- dependent aerobic glycolysis, i.e. the Warburg Effect, which is a common feature of cancer cells. We propose to study how fuel choice controls MondoA transcriptional activity in vitro and in vivo and how signals from the tumor microenvironment dictate MondoA transcriptional activity. In Aim 1, we will determine how TXNIP, hypoxia and how the balance between glycolysis and oxidative phosphorylation controls MondoA transcriptional activity. In Aim 2, we will determine whether MondoA is required for the establishment or maintenance of tumorigenesis and whether MondoA is transcriptionally active in well perfused/oxygenated tumor regions. To examine the generality of our findings, we will determine MondoA’s role in gene expression and nutrient-sensing using other breast cancer cell lines and in conditionally reprogrammed breast cancer cells from PDX models. In Aim 3, we will use genomics approaches to discover the direct MondoA-dependent transcriptome in vivo and determine how these genes collaborate with MondoA to sustain TNBC tumorigenesis. We will also discover the cofactors required for MondoA transcriptional activity in vivo using biotin proximity labeling.

摘要乳腺癌是一种异质性疾病，由多种临床和分子特征不同的疾病组成。与其他亚型相比，三阴性乳腺癌 (TNBC) 占所有亚型的 15-20%。乳腺癌的发病年龄较年轻，复发率较高，且生存率较低。我们的短期目标是剖析 TNBC 的分子途径。支持 TNBC 的发展和生存。我们的长期目标是利用这些发现来确定新的目标。适合治疗开发。我们专注于正常细胞和癌细胞如何整合。我们最感兴趣的是细胞外生长和营养信号如何维持生长和肿瘤发生。 Myc 转录因子扩展网络的两个成员整合了生长和营养信号。在本申请中，MondoA 的功能相反，可控制 TNBC 的侵袭性生长和存活。我们重点研究 MondoA 对基因表达的控制以及代谢如何塑造 MondoA 依赖性 MondoA 是葡萄糖依赖性转录的主要调节因子。硫氧还蛋白相互作用蛋白 (TXNIP) 是其最具特征的直接转录和葡萄糖诱导转录 TXNIP 具有多种功能，其中最突出的功能是燃料选择。 TXNIP 是葡萄糖摄取和有氧糖酵解的有效抑制剂，但它也可以驱动以下物质的分解代谢：因此，高 TXNIP 水平会驱动线粒体中的其他燃料，例如支链氨基酸和酮。线粒体中的燃料氧化，同时限制葡萄糖的离线利用，低 TXNIP 水平提示燃料的使用。我们已经证明，几种主要的癌基因会阻断葡萄糖和远离线粒体。 MondoA 转录活性，降低 TXNIP 水平，这可能有助于致癌基因-。我们提出，依赖性有氧糖酵解，即瓦伯格效应，这是癌细胞的一个共同特征。研究燃料选择如何控制 MondoA 体外和体内转录活性以及来自肿瘤微环境决定 MondoA 转录活性在目标 1 中，我们将确定 TXNIP、缺氧以及糖酵解和氧化磷酸化之间的平衡如何控制 MondoA 在目标 2 中，我们将确定 MondoA 是否是建立所必需的。肿瘤发生的维持以及 MondoA 在灌注/充氧良好的情况下是否具有转录活性为了检验我们研究结果的普遍性，我们将确定 MondoA 在基因表达中的作用。使用其他乳腺癌细胞系和条件重编程乳腺癌细胞进行营养感应在目标 3 中，我们将使用基因组学方法来发现直接的 MondoA 依赖性。体内转录组并确定这些基因如何与 MondoA 合作维持 TNBC 我们还将利用体内方法发现 MondoA 转录活性所需的辅助因子。生物素邻近标记。