Regulation of Cellular Proliferation by Novel Mitochondrial-Encoded Tumor Suppressors

新型线粒体编码肿瘤抑制剂对细胞增殖的调节

• 批准号: 10238768
• 负责人: Changhan Lee
• 金额: $ 32.69万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238768
• 关键词: ATAC-seq; Adenosine Monophosphate; Affect; Amino Acids; Bacteria; Basic Science; Binding; Bioinformatics; Cancer Model; Cell Cycle; Cell Nucleus; Cell Proliferation; Cell Separation; Cell Survival; Cell physiology; Cells; Cellular Stress; Chromatin; Code; Communication; Computer Analysis; Computing Methodologies; Coupled; DNA; DNA Binding; Data; Developmental Therapeutics Program; Electron Transport; Endothelium; Eukaryota; Eukaryotic Cell; Evolution; Exhibits; FDA approved; Foundations; Gene Expression; Genes; Genetic; Genetic Diseases; Genome; Genomic approach; Genomics; Homeostasis; In Vitro; Insulin Resistance; Language; Longevity; Machine Learning; Malignant - descriptor; Malignant Neoplasms; Maps; Mediating; Metabolic; Metabolic stress; Metabolism; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Molecular Biology Techniques; Molecular Genetics; Molecular and Cellular Biology; Mus; Names; Normal Cell; Normal tissue morphology; Nuclear; Nuclear Translocation; Nutrient; Open Reading Frames; Organelles; Osteoporosis; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Process; Property; Protein Kinase; Proteins; Proteomics; Publishing; RNA; Regulation; Reporting; Resistance; Ribosomal RNA; Ribosomes; Role; Signal Transduction; Site-Directed Mutagenesis; Source; Stress; System; TP53 gene; Testing; The Cancer Genome Atlas; Time; Tissue Microarray; Tissues; Toxic effect; Treatment Efficacy; Tumor Suppressor Proteins; age related; antitumor effect; base; cancer cell; cancer genome; cancer type; cell growth regulation; cell type; chemotherapy; chromatin immunoprecipitation; combinatorial; comparative; diet-induced obesity; exhaustion; functional genomics; genomic data; immune function; improved; in vivo; loss of function; mitochondrial genome; mouse model; muscle metabolism; mutant; new therapeutic target; novel; prevent; response; sensor; single-cell RNA sequencing; therapeutic development; tumor

ABSTRACT Cellular compartments are coordinated through a dynamic bidirectional communication network amongst various organelles. Here, we focus on the communication between mitochondria and the nucleus, organelles that each possess their own genomes. The mitochondrial and nuclear genomes have co-evolved for over a billion years and have likely required close communication and cross-regulation. However, whereas mitochondria are known to be regulated by over 1,000 nuclear-encoded proteins, but there is currently no known mitochondrial-encoded factor that actively communicates to and regulates the nucleus. We have recently identified a novel gene encoded within the mitochondrial DNA and named it MOTS-c (Mitochondrial ORF within the Twelve S rRNA type-c). MOTS-c is a small 16 amino acid peptide that regulates metabolic homeostasis, in part, via the master nutrient sensor AMPK (adenosine monophosphate-activated protein kinase). We recently reported that MOTS-c can translocate into the nucleus in response to metabolic stress to bind to chromatin and regulate nuclear gene expression. Further, our preliminary study using a multi-pronged approach, including single cell RNA-seq, bioinformatics (including machine learning), chromatin immunoprecipitation (ChIP) coupled with quantitative PCR (qPCR), and cell sorting, showed that MOTS-c can regulate cellular proliferation; MOTS-c targeted the p53/p21 pathway and ribosomal processes. Considering the important metabolic role of mitochondria in cellular proliferation processes (29), a critical question that remains largely enigmatic is how mitochondrial-encoded factors communicate to the nucleus to coordinate the metabolic shift with gene expression during proliferation. Notably, rapidly dividing cancer cells had undetectable levels of MOTS-c or nuclear-translocation deficiency, suggesting loss of mito-nuclear communication by MOTS-c. Together, cancer may be a genetic disease in which our two genomes exist in a state of disrupted bi-directional communication/regulation, and may serve as a unique model to start understanding the role of MOTS-c in cellular proliferation. Because MOTS-c expression/function was dysregulated and that MOTS-c can negatively regulate cell cycle/proliferation, we hypothesize that MOTS-c is a mitochondrial-encoded tumor suppressor, the first of its kind to be identified, that directly regulates the nucleus to coordinate cellular metabolism with proliferation. We propose three aims to test this hypothesis. First, we will characterize MOTS-c as a tumor suppressor that regulates cell proliferation at the molecular, cellular, genetic level. Second, we will comprehensively map the MOTS-c-dependent functional nuclear genomic landscape using multiple complimentary genomics approach, including single cell RNA-seq, ATAC-seq (chromatin accessibility), and genomic footprinting using ChIP-seq. The data from each genomic approach will be integrated using cutting-edge computational methods, including machine learning, to decipher the message(s) MOTS- c delivers to the nuclear genome to regulate cancer cell proliferation and survival. Lastly, we will determine how MOTS- c-mediated communication to the nucleus can differentially regulate cellular proliferation and stress resistance in normal and malignant cells using mouse models of cancer. If successful, we predict that our study will have broad and lasting impact on (i) basic research by introducing the paradigm-shifting concept of mitochondrial-encoded tumor suppressors that coordinate cellular metabolism and proliferation and (ii) therapeutic development by revealing mtDNA as a source of novel drug targets (currently there are no FDA-approved drugs based on the mitochondrial genome).

抽象的 蜂窝室通过各种动态双向通信网络协调 细胞器。在这里，我们专注于线粒体与细胞核之间的通信，每个细胞器 拥有自己的基因组。线粒体和核基因组共同发展了十亿年， 可能需要密切的沟通和交叉调节。但是，众所周知线粒体是 受1,000多个核编码蛋白的调节，但目前尚无已知线粒体编码因子 这积极通信并调节细胞核。我们最近确定了一个编码的新型基因 线粒体DNA并将其命名为MOTS-C（十二个s型C中的线粒体ORF）。 mots-c是a 小的16个氨基酸肽，可以通过主要营养传感器AMPK来调节代谢稳态 （腺苷一磷酸激活的蛋白激酶）。我们最近报告说MOTS-C可以转移到 核对代谢应激的响应核与染色质结合并调节核基因表达。此外，我们的 使用多管齐的方法的初步研究，包括单细胞RNA-seq，生物信息学（包括机器 学习），染色质免疫沉淀（芯片）与定量PCR（QPCR）和细胞分类相结合 MOTS-C可以调节细胞增殖。 MOTS-C针对p53/p21途径和核糖体过程。 考虑线粒体在细胞增殖过程中的重要代谢作用（29），这是一个关键的问题 这在很大程度上仍然是神秘的是线粒体编码的因素如何与细胞核交流以协调 代谢转移在增殖过程中与基因表达。值得注意的是，迅速分裂的癌细胞具有无法检测到的水平 MOTS-C或核转移缺陷的缺陷，表明MOTS-C失去了MITO-核通信。 总之，癌症可能是一种遗传疾病，其中我们的两个基因组以双向干扰状态存在 沟通/调节，可以作为开始理解MOTS-C在细胞中的作用的独特模型 增殖。因为MOTS-C表达/功能失调，并且MOTS-C可以负调节 细胞周期/增殖，我们假设MOTS-C是线粒体编码的肿瘤抑制剂，这是同类的第一个抑制剂 可以鉴定，直接调节细胞核以与增殖相协调细胞代谢。我们提出了三个 旨在检验这一假设。首先，我们将MOTS-C作为调节细胞增殖的肿瘤-C表征 分子，细胞，遗传水平。其次，我们将全面绘制MOTS-C依赖性功能核 使用多种免费基因组学方法的基因组景观，包括单细胞RNA-Seq，ATAC-Seq （染色质的可及性）和使用CHIP-SEQ进行基因组足迹。每种基因组方法的数据将是 使用包括机器学习在内的尖端计算方法进行集成，以破译消息 C向核基因组传递以调节癌细胞的增殖和存活。最后，我们将确定如何mots- C介导的与细胞核的通信可以在正常情况下差异地调节细胞增殖和应激性抗性 使用癌症的小鼠模型和恶性细胞。 如果成功，我们预测我们的研究将对（i）基础研究产生广泛而持久的影响 线粒体编码的肿瘤抑制剂的范式转移概念，该肿瘤抑制剂协调细胞代谢和 通过揭示mtDNA作为新药物的来源（目前在那里），增殖和（ii）治疗性发育 不是基于线粒体基因组的FDA批准药物）。