Investigating Mechanisms of Deregulated Nucleotide Metabolism in Cancer

研究癌症中核苷酸代谢失调的机制

• 批准号: 10452714
• 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
• 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/10452714
• 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; rational design; single molecule; stoichiometry; therapeutically effective; tumor; tumor metabolism

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，许多癌症都通过细胞毒性化疗来治疗，这些化疗利用了这些癌症对药物的依赖性。 增强核苷酸生物合成。然而，使这些化合物如此的细胞毒性特性 有效杀死癌细胞也对正常增殖细胞和组织造成严重破坏。为了 为了了解如何更有效、更安全地利用这一漏洞，我们必须集中精力， 癌细胞而非正常细胞、增殖和存活特异性需要的靶标。我 已经发现了一种这样的靶-磷酸核糖焦磷酸合成酶2 （PRPS2）。PRPS 2及其同源物PRPS 1产生产生所有必需的关键前体， 核苷酸和功能作为一个“分子节流阀”能够增加或减少的速度，这些 基因构建模块被制造出来。这一建议试图解开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，和生物正交化学， 分子生物学方法。我们的研究将为理解代谢 癌症细胞的脆弱性，并可能导致未来智能设计的合理治疗策略。 我们将使用MYC驱动的淋巴瘤和骨髓瘤模型进行我们的研究，使用遗传- 工程小鼠模型和人类癌细胞系。重要的是，MYC被描述为 癌症的转录引擎及其刺激核苷酸和核酸产生的能力是标志性的 其促生长合成代谢程序的特征是在B细胞谱系中驱动恶性肿瘤所必需的。使用我们 基因方法阻断PRPS 2在MYC过表达细胞中的功能，我们可以利用这种依赖性 解读MYC过度表达癌症中核苷酸代谢失调的机制基础 细胞，并揭示核苷酸代谢网络中关键节点之间的新联系。为 例如，我们提出的研究将阐明核苷酸代谢的经济学，通过确定如何 中断核苷酸供应会影响它所驱动的机器，反之亦然。总体而言，拟议的研究 将改变我们对这些关键分子在正常和癌症中的作用的理解， 设置，并提供一个新的概念范式，可以为下一个发展的基础， 产生更安全、更有效的基于精确度的疗法和方法。