Investigating Mechanisms of Deregulated Nucleotide Metabolism in Cancer

研究癌症中核苷酸代谢失调的机制

基本信息

批准号：
10225501
负责人：
Tom Cunningham
金额：
$ 36.71万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10225501
关键词：
Affect Amino Acid Substitution Anabolism Apoptosis B-Lymphocytes Biochemical Biochemistry CRISPR/Cas technology Cancer Model Cancer cell line Cell Lineage Cell Proliferation Cell Survival Cells Chemistry Chimerism Complex Cre-LoxP Cytosol Cytotoxic Chemotherapy DNA Polymerase I DNA Polymerase II DNA Polymerase III DNA-Directed RNA Polymerase Data Dependence Development Diphosphates Economics Enzymatic Biochemistry Enzymes Feedback Foundations Future Gene Expression Genetic Genetic Transcription Genetically Engineered Mouse Growth Hematopoietic Homo Homologous Gene Human Immunologics Individual Intelligence Knockout Mice Lead Lesion Link Lymphoma Lymphomagenesis MYC gene Malignant Neoplasms Metabolic Metabolism Microscopy Modeling Molecular Molecular Biology Multienzyme Complexes Multiple Myeloma Mus Natural regeneration Normal Cell Normal tissue morphology Nucleic Acids Nucleotide Biosynthesis Nucleotides Oncogenic Output Pathway interactions Patients Pentosephosphate Pathway Phenotype Physiology Production Proliferating Property Purines Pyrimidine RNA chemical synthesis Refractory Research Resolution Ribose Ribose-Phosphate Pyrophosphokinase Role Route Stable Isotope Labeling Structural Models Structure Testing Therapeutic Tissues Toxic effect Work anti-cancer base c-myc Genes cancer cell cytotoxic design enzyme activity genetic approach inorganic phosphate liquid chromatography mass spectrometry loss of function mouse model mutant next generation novel nucleic acid biosynthesis nucleotide metabolism overexpression programs pyridine single molecule stoichiometry therapeutically effective tumor tumor metabolism

项目摘要

PROJECT SUMMARY Many cancers are currently treated by cytotoxic chemotherapies that exploit those cancers' dependence on enhanced nucleotide biosynthesis. However, the cytotoxic properties which make these compounds so efficacious in killing cancer cells also wreak havoc on normal proliferating cells and tissues. In order to understand how to exploit this vulnerability more effectively and more safely, we must focus our efforts on targets that are specifically required by cancer cell, but not normal cell, proliferation and survival. My discoveries have identified one such target – the enzyme phosphoribosyl pyrophosphate synthetase 2 (PRPS2). PRPS2, and its homolog PRPS1, generate a critical precursor necessary for producing all nucleotides and function as a `molecular throttle' capable of increasing or decreasing the rate at which these genetic building blocks are made. This proposal seeks to unravel the molecular basis for this selectivity through use of metabolic flux analysis, elegant structure/function studies, and bioorthogonal chemistry and molecular biology approaches. Our studies will open up new avenues for understanding the metabolic vulnerabilities of cancer cells and may lead to intelligently-designed rational therapeutic strategies of the future. We will conduct our studies using models of MYC-driven lymphoma and myeloma, using both genetically- engineered mouse models and human cancer cell lines. Importantly, MYC has been characterized as the transcriptional engine of cancer and its ability to stimulate nucleotide and nucleic acid production are signature features of its pro-growth anabolic program necessary to drive malignancies in the B cell lineage. Using our genetic approaches that block PRPS2 function in MYC overexpressing cells, we can leverage this dependency to decipher the mechanistic basis for the deregulation of nucleotide metabolism in MYC-overexpressing cancer cells and uncover novel connections between critical nodes in the nucleotide metabolism network. For example, our proposed studies will elucidate the economics of nucleotide metabolism by determining how disrupting nucleotide supply affects the machineries it fuels, and vice-versa. Collectively, the proposed studies will be transformative in our understanding of the roles of these key molecules in the normal and cancer setting, and provide a new conceptual paradigm which can be the foundation for the development of the next generation of safer, more effective precision-based therapies and approaches.

项目摘要目前，许多癌症都受到细胞毒性化学疗法的治疗，这些化学疗法利用这些癌症对这些癌症的依赖增强的核卫星生物合成。但是，使这些化合物的细胞毒性特性如此为了杀死癌细胞，也会对正常的增殖细胞和组织造成严重破坏。了解如何更有效，更安全地利用这种脆弱性，我们必须将精力集中在癌细胞特别需要的靶标，但不是正常细胞，增殖和存活。我的发现已经确定了一个这样的靶标 - 磷酸磷酸酶合成酶2 （PRPS2）。 PRPS2及其同源性PRPS1产生了生产所有人所必需的关键先驱核苷酸和功能作为“分子节气门”，能够增加或降低这些速率建立了遗传基础。该提议旨在阐明这种选择性的分子基础通过使用代谢通量分析，优雅的结构/功能研究以及生物正交化学和分子生物学方法。我们的研究将为理解新陈代谢开辟新的途径癌细胞的脆弱性，并可能导致未来智能设计的理性治疗策略。我们将使用MYC驱动的淋巴瘤和骨髓瘤模型进行研究，这两者在遗传学上 - 工程的小鼠模型和人类癌细胞系。重要的是，MYC已被描述为癌症的转录发动机及其刺激核苷酸和核酸产生的能力是特征在B细胞谱系中驱动恶性肿瘤所必需的促增长合成代谢程序的功能。使用我们的阻断MYC过表达细胞中PRPS2功能的遗传方法，我们可以利用这种依赖性解释了在过表达MYC的癌症中放松核苷酸代谢的机理基础细胞并发现核苷酸代谢网络中关键节点之间的新型连接。为了例如，我们提出的研究将通过确定如何确定如何阐明核苷酸代谢的经济学破坏核丁基供应会影响其燃料的机械，反之亦然。拟议的研究集体我们对这些关键分子在正常和癌症中的作用的理解将具有变革性设置并提供一个新的概念范式，这可以成为下一个开发的基础产生更安全，更有效的基于精确的疗法和方法。