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

基本信息

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
项目类别：
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

项目摘要

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.

在我们之前的研究中，我们已经证明砷 (As3+)，一种环境类金属，能够诱导人支气管上皮细胞转化的其他实验数据揭示了其存在。其他初步数据显示，As3+ 诱导的转化细胞中存在癌症干样细胞 (CSC)。 (i)连续低浓度As3+处理人支气管上皮诱导的CSCs 由于严重抑制活性氧，细胞表现出活性氧化物质（ROS）显着减少。线粒体氧化磷酸化 (OXPHOS)；(ii) As3+ 诱导的 JNK 和 STAT3 (pSTAT3S727) 线粒体磷酸化以及线粒体转录因子 A (TFAM) 减少；综合转录组学和代谢组学分析表明糖酵解率较高且水平较低 As3+ 诱导的 CSC 中线粒体 DNA (mtDNA) 耗竭导致线粒体代谢的变化； (iv) 与幼稚胚胎干细胞 (ESC) 和癌细胞不同的独特糖酵解特征在这些 As3+ 诱导的 CSC 中发现，ESC 和癌细胞都直接糖酵解以产生乳酸。相比之下，As3+诱导的CSC显示糖酵解中间体向辅助物质的转化增加 N-乙酰氨基葡萄糖的生成途径对干性基因的 O-GlcNAc 酰化很重要 S-腺苷甲硫氨酸 (SAM) 有助于 DNA 和组蛋白甲基化。应用的目的是确定：(1) As3+ 诱导的 JNK 依赖性 pSTAT3S727 是否负责抑制线粒体；(2) 如果是这样，As3+ 引发的 JNK 依赖性 pSTAT3S727 信号通路如何损害线粒体线粒体的完整性或功能，例如 mtDNA 复制、转录、OXPHOS 等；以及 (3) 线粒体功能受损有助于 As3+ 诱导 CSC 的产生。 As3+ 诱导的 JNK 依赖性 pSTAT3S727 信号通过抑制线粒体 OXPHOS 和 ROS 的产生，以及随后细胞糖酵解的增强。检验这一假设，提出以下三个具体目标：具体目标 1：确定 As3+ 激活线粒体 JNK，磷酸化 BEAS-2B 和其他肺细胞中的 STAT3 S727 (pSTAT3S727) 我们将重点关注 JNK 线粒体局部上游激酶的激活。将研究其他线粒体蛋白对 As3+ 的 JNK 依赖性磷酸化反应。通过线粒体磷酸化蛋白质组；具体目标2：了解As3+如何诱导JNK依赖性 pSTAT3S727 通过解决 pSTAT3S727 在 mtDNA 结合中的作用及其与线粒体的相互作用来抑制线粒体线粒体转录因子 A (TFAM) 及其对线粒体 ROS 产生、蛋白质组和线粒体的影响 As3+ 处理的细胞和 As3+ 诱导的 CSC 的细胞代谢组学；具体目标 3：利用我们独特的小鼠原位肺癌模型与临床相关靶向剂一起进行临床前探索 JNK、STAT3 和糖酵解抑制剂在 CSC 生成的小鼠肺癌模型中的治疗潜力。我们将测试 JNK、STAT3 和糖酵解抑制剂的短期和长期全身治疗方案。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 影响 CSC 的生成并导致 As3+ 致癌的新概念的出现此外，我们相信 As3+ 在 CSC 诱导中的能力是从完成时生成的。这个项目的真正价值在于定义一些新的、直接的目标点，这些目标点可能会有所帮助加速它们在临床环境中的使用。