Extracellular ATP in cancer: mechanism study of ATP internalization and ATP-induced drug resistance in cancer cells and tumors

癌症中的细胞外 ATP：癌细胞和肿瘤中 ATP 内化和 ATP 诱导的耐药性的机制研究

• 批准号: 9812230
• 负责人: Xiaozhuo Chen
• 金额: $ 45.05万
• 依托单位: OHIO UNIVERSITY ATHENS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-10 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9812230
• 关键词: A549; ATP-Binding Cassette Transporters; Address; Affect; Animals; Antibodies; Applications Grants; Biochemical Reaction; Biological; Biological Process; Cancer Cell Growth; Cancer Center; Cancer Patient; Cancer cell line; Cell Proliferation; Cell Survival; Cells; Cellular Metabolic Process; Coupled; Cytosol; Destinations; Development; Drug Targeting; Drug resistance; Endosomes; Energy Metabolism; Enzymes; Event; Free Energy; Genetic Transcription; Genetic study; Goals; Growth; Human; In Vitro; Knowledge; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Membrane; Modeling; Molecular; Mus; Names; Nature; Normal tissue morphology; Nutrient; Organism; Paper; Pathway interactions; Phenotype; Phosphorylation; Play; Prevalence; Process; Protein Analysis; Proteins; Publications; Radioactive; Reaction; Regulation; Reporting; Research; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Signaling Molecule; Source; Stromal Cells; Testing; Time; Training; Warburg Effect; Work; analog; anti-cancer therapeutic; base; bioluminescence imaging; cancer cell; cancer therapy; cancer type; cell type; design; effective therapy; extracellular; improved; in vivo; insight; novel; novel therapeutics; protein biomarkers; reverse genetics; targeted treatment; treatment strategy; tumor; tumor growth; tumor metabolism; tumor microenvironment; tumor xenograft; undergraduate student; uptake

ABSTRACT ATP is one of the most versatile biological molecules in all organisms. It provides readily available energy for almost all enzymatic reactions and phosphorylates proteins and enzymes in almost all signal transduction cascades. It also functions as an extracellular messenger. Although all of these biological processes cannot function without ATP, ATP has almost been taken for granted in performing these essential functions, and research focuses have been much more on other molecules involved in these reactions. Recent research has shown that ATP plays some very important but previously unrecognized functions in cancer. Intratumoral extracellular ATP (eATP) is found to be more than 1,000 times higher than that in normal tissues, in the range of 100-500 µM, but the sources, destination and functions of the high levels of eATP are still largely unclear. Our recent studies have revealed that eATP can function either extracellularly or intracellularly because eATP can be internalized by cancer cells by different types of endocytotic processes, particularly macropinocytosis. Once inside cells, ATP elevates intracellular ATP levels; promotes cell metabolism, growth and proliferation; enhances signal transduction for cell survival and proliferation; and augments drug resistance. However, exactly how eATP is internalized and then released by cancer cells is far from fully understood. It is also presently unknown if the eATP internalization is a general phenomenon and phenotype among different cancer types. Furthermore, how and if internalized eATP-induced drug resistance is regulated at transcriptional and translational levels is largely unclear. There is a big gap in our current understanding of eATP and our ultimate goal of more effectively treating cancer by disrupting eATP traffic and functions. The long-term research goal of my lab is to understand how eATP affects cancer at molecular, cellular and animal levels and use gained knowledge to design novel therapies to more effectively treat cancer. The objectives of this proposed study are to determine the prevalence and mechanisms that control and regulate ATP's cross-membrane internalization and intracellular release, and drug resistance in cancer. The central hypothesis of this study is that eATP is internalized by most cancer types and then released inside cells to perform diverse functions such as promoting cell survival and drug resistance not only at the signaling level but also at transcriptional and translational levels. To test this hypothesis, three specific aims are proposed: Specific Aim 1. Determine the prevalence of eATP internalization among difference cancer cell lines and cancer types. Specific Aim 2. Use various ATP analogs and endosome markers to study mechanisms of internalization and release of eATP in cancer cells. Specific Aim 3. Use reverse genetics and bioluminescence imaging to study eATP-mediated drug resistance in cancer cells and xenografted tumors in mice. The completion of the study will provide insights into how eATP gets into cancer cells and performs functions such as promotion of drug resistance, and identification of novel targets for reducing drug resistance and improving cancer patients' survival.

摘要 三磷酸腺苷是所有生物中用途最广泛的生物分子之一。它提供了随时可用的能量， 几乎所有的酶反应和磷酸化几乎所有的信号转导中的蛋白质和酶 瀑布。它还起到细胞外信使的作用。尽管所有这些生物过程都无法 没有ATP的功能，在执行这些基本功能时，ATP几乎被认为是理所当然的，并且 研究的重点更多地放在参与这些反应的其他分子上。最近的研究表明 研究表明，ATP在癌症中发挥着一些非常重要但以前未被认识到的功能。瘤内 在这个范围内，细胞外的ATP(EATP)被发现比正常组织高出1000多倍。 100-500微米，但高水平eATP的来源、目的地和功能在很大程度上仍不清楚。 我们最近的研究表明，eATP可以在细胞外或细胞内发挥作用，因为eATP 可通过不同类型的内吞过程，特别是巨噬细胞吞噬作用被癌细胞内化。 一旦进入细胞，三磷酸腺苷就会提高细胞内的三磷酸腺苷水平，促进细胞新陈代谢、生长和增殖； 增强细胞生存和增殖的信号转导；并增强耐药性。然而， EATP究竟是如何内化，然后由癌细胞释放的，目前还远未完全了解。它也是 目前尚不清楚eATP内化是否是不同肿瘤之间的普遍现象和表型 类型。此外，内在化eATP诱导的耐药性如何以及是否在转录和转录水平上受到调节。 翻译水平在很大程度上还不清楚。我们目前对eATP的理解存在很大差距，我们最终的 通过扰乱eATP的交通和功能来更有效地治疗癌症的目标。长期的研究目标 我实验室的主要任务是了解eATP如何在分子、细胞和动物水平上影响癌症，并利用 设计新疗法以更有效地治疗癌症的知识。这项拟议研究的目标 是为了确定控制和调节ATP跨膜的流行率和机制 肿瘤的内化和细胞内释放，以及耐药。这项研究的中心假设是 EATP被大多数类型的癌症内化，然后在细胞内释放，以执行各种功能，如 AS不仅在信号水平上促进细胞存活和耐药性，而且在转录和 翻译级别。为了验证这一假设，提出了三个具体目标：具体目标1.确定 EATP内化在不同癌细胞系和癌症类型中的流行率。具体目标2.使用 不同的ATP类似物和内吞体标记物研究eATP内化和释放的机制 癌细胞。具体目的3.利用反向遗传学和生物发光成像技术研究eATP介导的 小鼠癌细胞和异种移植瘤的耐药性。研究完成后，将会提供 深入了解eATP是如何进入癌细胞并发挥促进耐药性等功能的， 并确定减少耐药性和提高癌症患者生存的新靶点。