Reduced Reactive Oxygen Species and Oxidative Phosphorylation in Arsenic-Induced Cancer Stem Cells

砷诱导的癌症干细胞中活性氧的减少和氧化磷酸化

• 批准号: 10441939
• 负责人: Fei Chen
• 金额: $ 35.55万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-14 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10441939
• 关键词: Address; Aerobic; Area; Argentina; Arsenic; Bangladesh; Biochemical; Bladder; Cancer Model; Carcinogens; Cell Line; Cells; Characteristics; Chemicals; Chile; China; Clinical; Cytoplasm; DNA Binding; DNA Methylation; DNA biosynthesis; Data; Deposition; Environmental Exposure; Epidemiology; Epithelial Cells; Etiology; Exhibits; Exposure to; Generations; Genes; Genetic Transcription; Glycolysis; Goals; Human; Impairment; India; International Agency for Research on Cancer; JNK-activating protein kinase; Lead; Link; Liver; Lung; MAPK8 gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Metabolic; Metabolism; Metals; Mexico; Mitochondria; Mitochondrial DNA; Mitochondrial Proteins; Molecular; Mongolia; Mus; NOD/SCID mouse; Oxidative Phosphorylation; Pathway interactions; Phosphorylation; Planet Earth; Population; Preclinical Testing; Production; Prostate; Proteome; Public Health; Reactive Inhibition; Reactive Oxygen Species; Regimen; Research; Role; S-Adenosylhomocysteine; STAT3 gene; Serine; Signal Pathway; Signal Transduction; Skin; Taiwan; Testing; Therapeutic; alkalinity; arsenic carcinogenesis; base; bronchial epithelium; cancer cell; cancer stem cell; cancer therapy; carcinogenesis; carcinogenicity; cell transformation; clinically relevant; drinking water; embryonic stem cell; exposed human population; ground water; histone methylation; inhibitor/antagonist; metabolomics; mitochondrial metabolism; mouse model; mtTF1 transcription factor; pre-clinical; response; stem-like cell; stemness; targeted agent; targeted treatment; therapeutic evaluation; transcriptomics; tumor; upstream kinase; Address; Aerobic; Area; Argentina; Arsenic; Bangladesh; Biochemical; Bladder; Cancer Model; Carcinogens; Cell Line; Cells; Characteristics; Chemicals; Chile; China; Clinical; Cytoplasm; DNA Binding; DNA Methylation; DNA biosynthesis; Data; Deposition; Environmental Exposure; Epidemiology; Epithelial Cells; Etiology; Exhibits; Exposure to; Generations; Genes; Genetic Transcription; Glycolysis; Goals; Human; Impairment; India; International Agency for Research on Cancer; JNK-activating protein kinase; Lead; Link; Liver; Lung; MAPK8 gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Metabolic; Metabolism; Metals; Mexico; Mitochondria; Mitochondrial DNA; Mitochondrial Proteins; Molecular; Mongolia; Mus; NOD/SCID mouse; Oxidative Phosphorylation; Pathway interactions; Phosphorylation; Planet Earth; Population; Preclinical Testing; Production; Prostate; Proteome; Public Health; Reactive Inhibition; Reactive Oxygen Species; Regimen; Research; Role; S-Adenosylhomocysteine; STAT3 gene; Serine; Signal Pathway; Signal Transduction; Skin; Taiwan; Testing; Therapeutic; alkalinity; arsenic carcinogenesis; base; bronchial epithelium; cancer cell; cancer stem cell; cancer therapy; carcinogenesis; carcinogenicity; cell transformation; clinically relevant; drinking water; embryonic stem cell; exposed human population; ground water; histone methylation; inhibitor/antagonist; metabolomics; mitochondrial metabolism; mouse model; mtTF1 transcription factor; pre-clinical; response; stem-like cell; stemness; targeted agent; targeted treatment; therapeutic evaluation; transcriptomics; tumor; upstream kinase

In our previous studies we had demonstrated that arsenic (As3+), an environmental metalloid metal, was able to induce transformation of the human bronchial epithelial cells. Additional experimental data revealed presence of cancer stem-like cells (CSCs) among the transformed cells induced by As3+. Other preliminary data showed that: (i) The CSCs induced by consecutive low-concentration As3+ treatment of the human bronchial epithelial cells exhibited significant decrease of reactive oxidative species (ROS) due to severe inhibition of the mitochondrial oxidative phosphorylation (OXPHOS); (ii) As3+ induced JNK and STAT3 (pSTAT3S727) phosphorylation in mitochondria along with a diminish of the mitochondrial transcription factor A (TFAM); (iii) integrated transcriptomic and metabolomic analyses demonstrated a higher rate of glycolysis and lower levels of mitochondrial metabolism due to mitochondrial DNA (mtDNA) depletion among these As3+-induced CSCs; and (iv) a unique glycolytic feature that is different from naïve embryonic stem cells (ESCs) and cancer cells was found in these As3+-induced CSCs. Both ESCs and cancer cells direct glycolysis for lactate production. In contrast, the As3+-induced CSCs show increased conversion of the glycolytic intermediates into the subsidiary pathways for the generation of N-acetylglucosamine important for O-GlcNAcylation of the stemness genes and the S-adenosyl methionine (SAM) that contributes to DNA and histone methylation. Accordingly, the goal of this application is to determine: (1) is As3+-induced JNK-dependent pSTAT3S727 responsible for the inhibition of mitochondria; (2) if so, how this JNK-dependent pSTAT3S727 signaling pathway elicited by As3+ impairs the integrity or function of mitochondria, such as mtDNA replication, transcription, OXPHOS, etc; and (3) how the impaired function of mitochondria contributes to the generation of the CSCs induced by As3+. We hypothesize that As3+-induced JNK-dependent pSTAT3S727 signaling promotes formation of the CSCs by inhibiting mitochondrial OXPHOS and ROS generation, and the subsequent enhancement of glycolysis of the cells. To test this hypothesis, the following three specific aims are proposed: Specific Aim 1: determine how As3+ activates mitochondrial JNK that phosphorylates STAT3 S727 (pSTAT3S727) in BEAS-2B and other lung cells for the formation of CSCs. We will focus on the activation of mitochondrial-localized upstream kinases of JNK in response to As3+. The JNK dependent phosphorylation of additional mitochondrial proteins will be investigated through mitochondrial phosphoproteome; Specific aim 2: understand how As3+-induced JNK-dependent pSTAT3S727 inhibits mitochondria by addressing the role of pSTAT3S727 in mtDNA binding, its interaction with the mitochondrial transcription factor A (TFAM), and its effects on mitochondrial ROS production, proteome and the cellular metabolomics in the As3+-treated cells and As3+-induced CSCs; Specific Aim 3: utilize our unique mouse orthotopical lung cancer model, together with clinically relevant targeting agent, to preclinically explore therapeutic potential of inhibitors of JNK, STAT3 and glycolysis in CSC-generated lung cancer model in mice. Both short- and long-term systemic regimens of JNK, STAT3 and glycolysis inhibitors will be tested. We anticipate that the results from the proposed studies will unravel importance of As3+-induced JNK-dependent pSTAT3S727 on the generation of CSCs and lead to emerging of new concepts of As3+ carcinogenesis by emphasizing the ability of As3+ in CSC induction. Moreover, we believe that the date generated from completion of this project will be of real value in defining some new and straightforward targeting points that may help accelerate their use in clinical settings.

在以前的研究中，我们证明了砷（AS3+）是一种环境金属金属，能够 诱导人支气管上皮细胞的转化。其他实验数据揭示了存在 由AS3+诱导的转化细胞中的癌症干细胞（CSC）。显示的其他初步数据 那就是：（i）由人支气管上皮的连续低浓度AS3+治疗引起的CSC 由于严重抑制，细胞暴露了反应性氧化物物种（ROS）的显着降低 线粒体氧化磷酸化（OXPHOS）； （ii）AS3+诱导JNK和STAT3（PSTAT3S727） 线粒体中的磷酸化以及线粒体转录因子A（TFAM）的减少； （iii） 综合转录组和代谢组学分析表明，糖酵解的速率更高，较低的水平 这些AS3+诱导的CSC中线粒体DNA（mtDNA）部署引起的线粒体代谢； （iv）与幼稚的胚胎干细胞（ESC）和癌细胞不同的独特糖酵解特征是 在这些AS3+诱导的CSC中发现。 ESC和癌细胞都将糖酵解用于乳酸产生。在 对比，AS3+诱导的CSC显示出糖酵解中间体转化为子公司的增加 生成N-乙酰葡萄糖的途径，对干性基因的O-Glcnacylation重要 有助于DNA和组蛋白甲基化的S-腺苷（SAM）。根据这个目标 应用是确定：（1）AS3+诱导的JNK依赖性PSTAT3S727负责抑制 线粒体； （2）如果是这样，则该JNK依赖性PSTAT3S727信号传导途径是AS3+引起的 线粒体的完整性或功能，例如mtDNA复制，转录，oxphos等； （3）如何 线粒体的功能受损有助于AS3+诱导的CSC的产生。我们假设 AS3+诱导的JNK依赖性PSTAT3S727信号传导通过抑制促进CSC的形成 线粒体的Oxphos和ROS的产生，以及随后增强细胞糖酵解。到 检验该假设，提出以下三个特定目的：特定目的1：确定AS3+的方式 激活线粒体JNK，该JNK磷酸化STAT3 S727（PSTAT3S727）在BEAS-2B和其他肺部细胞中的磷酸化JNK CSC的形成。我们将重点介绍JNK的线粒体 - 区域化上游激酶的激活 对AS3+的响应。将研究其他线粒体蛋白的JNK依赖性磷酸化 通过线粒体磷酸蛋白酶；特定目标2：了解AS3+诱导的JNK依赖性如何 PSTAT3S727通过解决PSTAT3S727在mtDNA结合中的作用来抑制线粒体，并与之相互作用 线粒体转录因子A（TFAM）及其对线粒体ROS的影响，蛋白质组和 AS3+处理的细胞和AS3+诱导的CSC中的细胞代谢组学；特定目标3：利用我们独特的鼠标 矫形肺癌模型以及临床相关的靶向剂，以临床前探索 小鼠CSC生成的肺癌模型中JNK，STAT3和糖酵解抑制剂的治疗潜力。 将测试JNK，STAT3和糖酵解抑制剂的短期和长期全身性方案。我们 预计拟议研究的结果将揭示AS3+诱导的JNK依赖性的重要性 PSTAT3S727关于CSC的产生，并通过 强调AS3+在CSC诱导中的能力。此外，我们相信完成的日期 在定义一些可能有帮助的新的，直接的目标点方面，这个项目将具有真正的价值 加速其在临床环境中的使用。