Roles for Intracellular pH Dynamics in Cancer Cell Behaviors

细胞内 pH 动态在癌细胞行为中的作用

• 批准号: 10407970
• 负责人: Bree K. Grillo-Hill
• 金额: $ 10.99万
• 依托单位: SAN JOSE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-11 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407970
• 关键词: 3-Dimensional; Acids; Affect; Amino Acids; Animal Model; Antibodies; Apoptosis; Apoptotic; Area; Biology; Biosensor; Carrier Proteins; Cell Count; Cell Cycle; Cell Cycle Kinetics; Cell Cycle Progression; Cell Cycle Regulation; Cell Cycle Stage; Cell Death; Cell membrane; Cell physiology; Cells; Cellular Assay; Cellular biology; Complex; Coupled; Cues; Cyclin B; Data; Development; Disease; Disease Progression; Drosophila genus; Drosophila melanogaster; Dysplasia; Epidermal Growth Factor; Eye; G2 Phase; Genetic; Genetic Models; Genetic Screening; Goals; Growth; Human; Image; Imaging Techniques; Individual; Invaded; Ion Transport; Lead; Link; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Mediating; Methods; Mitochondria; Molecular; Molecular Analysis; NHE1; Neoplasm Metastasis; Oncogenes; Oncogenic; Outcome; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Patients; Phenotype; Phosphotransferases; Physiological; Post-Translational Protein Processing; Process; Protein Conformation; Proteins; Protons; Publishing; Reagent; Regulation; Research; Role; Signal Pathway; Signaling Molecule; Sodium; Stress; TP53 gene; Technology; Testing; Tissues; Tumor-Derived; cancer cell; cell behavior; cell growth; cell motility; fly; genetic analysis; in vivo; insight; migration; mutant; neoplastic cell; new therapeutic target; novel therapeutic intervention; overexpression; protein function; protonation; quantitative imaging; response; sensor; tool; tumor; tumor heterogeneity; tumor progression; tumorigenesis

Project Summary Constitutively increased intracellular pH (pHi) is common to most cancers regardless of tissue origin or genetic background. Increased pHi is necessary for survival of patient-derived tumor cells, and is sufficient to induce oncogenic phenotypes, including dysregulated tissue growth, dysplasia, and invasive cell migration. pH-regulated cell behaviors are predominantly mediated by changes in the protonation state of pH sensitive proteins, or pH sensors, which is viewed as a post-translational modification which cannot be resolved by mass spectrometry or antibodies. At the molecular level, changes in pHi can alter the protonation state of selective amino acid residues with pKa values near neutral, which can markedly affect protein conformations and function. By changing the protonation of multiple proteins in unison, pHi dynamics can coordinate complex cell processes. Although understudied, emerging evidence suggests that the higher pHi of most cancers contributes to pathogenesis. In this proposal, we will characterize pH sensitive cancer cell behaviors and identify pH sensors that regulate proliferation (Aim 1), programmed cell death (Aim 2) and metastasis (Aim 3). Using the genetic model organism Drosophila, we will bridge scales from tissue-level phenotypic analyses to individual pH-sensitive proteins that regulate these cancer cell behaviors. We will use cutting edge technologies including fluorescent cell cycle and apoptosis markers, genetically encoded pH bio-sensors, and quantitative imaging techniques to generate 3-D maps of intratumoral pHi. Our unpublished data with genetic modifier screens identified a number of candidate pH sensors with known roles in regulating these processes. We will test predictions on the pH-sensitive function of these candidate pH sensors. Significant outcomes from our studies include new insights on how pH-regulated cellular behaviors enable tumorigenesis and metastasis. The strengths of the PI in linking tissue-level phenotypes to individual pH-sensitive molecules permit unique insights in this under-studied area of biology. Further, understanding the molecular mechanisms of pH-sensitive proteins will inform novel therapeutic approaches to limit cancer progression.

项目摘要 组成性增加的细胞内pH（pHi）对于大多数癌症是常见的， 组织起源或遗传背景。增加的pHi对于患者源性肿瘤的存活是必要的。 肿瘤细胞，并足以诱导致癌表型，包括失调的组织 生长、发育不良和侵入性细胞迁移。pH调节的细胞行为主要是 由pH敏感蛋白或pH传感器的质子化状态的变化介导， 被视为不能通过质谱法解析的翻译后修饰，或 抗体的在分子水平上，pH的变化可以改变选择性质子化的质子化状态。 pKa值接近中性的氨基酸残基对蛋白质的影响显著 构象和功能。通过一致改变多种蛋白质的质子化，pHi 动力学可以协调复杂的细胞过程。虽然研究不足，但新的证据 表明大多数癌症的较高pHi有助于发病机制。 在这项提案中，我们将描述pH敏感的癌细胞行为，并确定pH 调节增殖（Aim 1）、程序性细胞死亡（Aim 2）和转移（Aim 3）。使用遗传模式生物果蝇，我们将从组织水平的桥梁尺度 对调节这些癌细胞行为的单个pH敏感蛋白进行表型分析。 我们将使用包括荧光细胞周期和凋亡标记在内的尖端技术， 基因编码的pH生物传感器，以及生成3D地图的定量成像技术 肿瘤内pHi的变化。我们未发表的遗传修饰筛选数据确定了一些 候选pH传感器在调节这些过程中具有已知的作用。我们将测试预测， 这些候选pH传感器的pH敏感功能。 我们研究的重要成果包括对pH调节细胞的新见解， 行为使肿瘤发生和转移。PI在连接组织水平方面的优势 表型到单个pH敏感分子允许在这一研究不足的领域的独特见解 生物学。此外，了解pH敏感蛋白的分子机制将有助于 限制癌症进展的新治疗方法。