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Arsenic-induced miRNA-199 and miRNA-214 deplete mitochondrial DNA for the generation of cancer stem-like cells

砷诱导的 miRNA-199 和 miRNA-214 消耗线粒体 DNA，从而产生癌症干细胞样细胞

基本信息

批准号：
9521211
负责人：
Fei Chen
金额：
$ 26.36万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9521211
关键词：
Area Arsenic Binding Biochemical Bladder CRISPR/Cas technology Cancer Model Carcinogens Case-Control Studies Cell Line Cells ChIP-seq Characteristics Chromatin Chromatin Structure Clinical DNA Methylation DNA biosynthesis Data Deposition Dose Environmental Exposure Epigenetic Process Epithelial Cells Exhibits Exposure to Generations Genes Genetic Genetic Transcription Geology Glucose Glycolysis Goals Histone H3 Human Impairment In Vitro International Agency for Research on Cancer Lead Link Liver Lung Lysine MAPK8 gene Malignant - descriptor Malignant Neoplasms Malignant neoplasm of lung Maps Metabolic Metabolic Pathway Metabolism Metals MicroRNAs Mitochondria Mitochondrial DNA Molecular Monitor Mus NOD/SCID mouse Non-Small-Cell Lung Carcinoma Organ Oxidative Phosphorylation Pathway interactions Phosphorylation Planet Earth Preclinical Testing Production Prostate Public Health Reporting Research Research Project Grants Risk Role S-Adenosylhomocysteine STAT3 gene Serine Signal Transduction Skin Testing Transcriptional Regulation base c-myc Genes cancer cell cancer stem cell cancer therapy carcinogenesis carcinogenicity cell transformation drinking water embryonic stem cell exposed human population ground water histone methylation in vivo knock-down metabolomics mitochondrial dysfunction mitochondrial metabolism mouse model mtTF1 transcription factor notch protein novel therapeutics overexpression pluripotency population based promoter response self-renewal stem-like cell stemness targeted treatment transcription factor transcriptome sequencing transcriptomics tumor

项目摘要

We have shown that consecutive treatment of the human bronchial epithelial cells with the environmentally relevant concentration of As3+ (0.125 – 0.25M), an environmental metalloid metal, for six months, induces transformation of the human bronchial epithelial cells, some of which possess characteristics of the cancer stem-like cells (CSCs), such as tumor sphere formation in vitro, self- renewal in vivo, increased expression of the stemness genes, including Oct4, Sox2, KLF4, and c-myc. In addition, these cancer stem-like cells exhibited a pronounced increase in the expression of several microRNAs, most notably, the miR-214, miR-199, miR-10b, miR-34b, etc. Furthermore, 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. Lastly, 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 miRNAs, esp. miR214/199, responsible for the depletion of mtDNA and the consequent inhibition of mitochondria; (2) if so, how miRNAs induced by As3+ impairs the integrity and function of mtDNA and mitochondria; 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 and miR-214/199 switch mitochondrial OXPHOS to glycolysis for the formation of CSCs. To test this hypothesis, the following three specific aims are proposed: Specific Aim 1: As3+-activated JNK and pSTAT3S727 enforce expression of miR-214 and miR-199 that down-regulate mitochondrial transcription factor A (TFAM) in BEAS-2B and other lung cells for the generation of CSCs. We will focus on the transcriptional regulation of the miR-214/199 cluster with emphases on promoter DNA methylation and transcription factor binding in cellular response to As3+ and its down-stream signaling; Specific aim 2: Understand how As3+-induced JNK, miR-214/199 and mitochondrial dysfunction contribute to the formation of CSCs with an emphasis on metabolic reprogramming from OXPHOS to glycolysis. Specific Aim 3: Defining the causal roles of As3+-induced JNK- and miR-214/199-dependent metabolic reprogramming in the changes of epigenetics related to chromatin structure and accessibility that linked to self-renewal and/or differentiation of the As3+-induced CSCs through high-throughput profiling. We will identify metabolite- dependent epigenetic and chromatin changes in non-transformed cells, As3+-induced transformed cells and CSCs, which will be further verified through overexpressing or CRISPR-Cas9-based knockdown of the key genes in the related metabolic pathways and monitoring the self-renewal and differentiation status of the CSCs. The high-throughput approaches will include ChIP-seq to map H3K9me3, H3K27me3, and H3K4me3, and RNA-seq to profile transcription of the genes, esp. for those contributing to the pluripotency, self-renewal and differentiation of the CSCs. We anticipate that the results from the proposed studies will unravel importance of As3+-induced miR-214/199 on the generation of CSCs and lead to emerging of new concepts of As3+ carcinogenesis by emphasizing the capability of As3+ in CSC induction. Moreover, we believe that the date generated from this project will help us in developing novel therapeutic strategies by targeting JNK, miR-214/199 and CSCs through utilizing our unique mouse orthotopical lung cancer model in NOD/SCID mice in a separate research project.

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