Investigating the role of DUX and miR-34a regulation in cell fate plasticity and cancer

研究 DUX 和 miR-34a 调节在细胞命运可塑性和癌症中的作用

• 批准号: 10518393
• 负责人: Bradley Don Weaver
• 金额: $ 4.82万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-11-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10518393
• 关键词: 3' Untranslated Regions; B-Cell Acute Lymphoblastic Leukemia; B-Lymphocyte Subsets; Binding Sites; Biological Assay; Cancer Biology; Cell Cycle; Cell Fate Control; Cell Proliferation; Cell Therapy; Cell model; Cells; Clinical; Collaborations; DNA Damage; Data; Development; Embryo; Embryonic Induction; Ensure; Epigenetic Process; Facioscapulohumeral Muscular Dystrophy; Family; Feedback; Gene Fusion; Genes; Genetic Transcription; Height; Human; Huntsman Cancer Institute at the University of Utah; Knock-out; Knockout Mice; Link; Luciferases; Malignant - descriptor; Malignant Neoplasms; Mammals; Maps; Measures; Mentorship; Messenger RNA; MicroRNAs; Molecular; Mus; Mutate; Mutation; Oncogenic; Pathway interactions; Physicians; Positioning Attribute; Post-Transcriptional Regulation; Probability; Productivity; Quantitative Reverse Transcriptase PCR; RNA; Regenerative Medicine; Regulation; Reporter; Repression; Role; S phase; Scientist; Site; Specific qualifier value; Surgeon; TP53 gene; Techniques; Teratoma; Testing; Therapeutic; Totipotency; Totipotent; Transcript; Translational Regulation; Translational Repression; Translations; Tumor Suppressor Proteins; Work; blastomere structure; cancer therapy; cell growth; cell regeneration; cell type; clinical development; embryo cell; embryo stage 2; embryo tissue; embryonic stem cell; epigenetic silencing; experimental study; homeodomain; human tissue; improved; in vivo; insight; mRNA Expression; novel; novel therapeutics; overexpression; posttranscriptional; prevent; programs; response; ribosome profiling; sarcoma; stem; stem cell fate; targeted cancer therapy; therapy resistant; transcription factor; transcriptome; tumorigenesis

Project Summary Hijacked plasticity of cell fate potential, combined with oncogenic mutations, drives malignant transformation and therapeutic resistance in many cancers. Totipotency – the ability of a cell to become all embryonic and extra- embryonic tissues – represents the height of embryonic/stem cell fate potential in mammals. DUX is a double homeodomain transcription factor that is misexpressed in subsets of B-cell acute lymphoblastic leukemia and round-cell sarcomas. Our lab recently defined DUX (mouse Dux / human DUX4) as a major driver of totipotent developmental programs in mouse and human. Pluripotent mouse embryonic stem cells (mESCs) expressing mouse Dux are potently converted into a ‘2C-like’ totipotent state, which epigenetically and transcriptionally resemble the 2-cell mouse embryo. In vivo, DUX expression is highly transient, and lasts less than a cell cycle in early embryos. Although the transcriptome of totipotency is well studied, it is unclear how the embryo employs post-transcriptional and translational regulation downstream of DUX to establish this plasticity in cell fate, and what mechanisms subsequently restrict developmental potency to ensure proper lineage determination. Recently, we and others have discovered a central role for p53 in DUX activation. P53 is a master tumor suppressor and transcription factor, which also activates the micro-RNA (miRNA) 34 family (a/b/c) in response to DNA damage. miR-34a is expressed in many mouse and human tissues, and is a well-studied tumor suppressor itself, repressing pathways involved in cellular growth and proliferation. Importantly, miR-34a deletion in mESCs confers a high probability of their conversion into a totipotent state, however the mechanisms governing this expanded cell fate plasticity are unclear. We find that loss of miR-34a in mESCs causes accumulation of Dux transcripts, and DUX4 contains a predicted miR-34a target site in its 3’ UTR, suggesting a direct regulatory role of miR-34a. We hypothesize that p53 and miR-34a activate and repress Dux (and/or Dux targets), respectively, forming a feedback loop which regulates the totipotency network and ultimately restricts cell fate plasticity in vivo. Uncovering the mechanisms safe-guarding cell fate potential and preventing oncogenesis in vivo will yield novel insights in both cancer biology and regenerative medicine. I am uniquely positioned to answer these questions in the Cairns Lab within the Huntsman Cancer Institute at the University of Utah. Leveraging our advanced DUX expertise, combined with key collaborations with experts in translational control, I am confident in my abilities to execute the experiments outlined within this proposal. Additionally, my excellent clinical mentorship team, crafted from highly productive physician- and surgeon-scientists, will promote my clinical development as I seek to combine key insights from molecular drivers of cell fate plasticity and oncogenesis into development of novel targets for cancer therapeutics and regenerative medicine.

项目摘要 细胞命运潜能的可塑性被劫持，与致癌突变相结合，驱动恶性转化， 许多癌症的治疗耐药性。全能性-一个细胞成为所有胚胎和额外的能力- 胚胎组织-代表哺乳动物中胚胎/干细胞命运潜力的高度。DUX是一个双 在B细胞急性淋巴细胞白血病亚群中错误表达的同源域转录因子， 圆细胞肉瘤我们的实验室最近将DUX（小鼠DUX /人DUX 4）定义为全能性的主要驱动因素。 小鼠和人类的发育程序。多能小鼠胚胎干细胞（mESC）表达 小鼠Dux被有效地转化为“2C样”全能状态， 类似于2细胞小鼠胚胎。在体内，DUX表达是高度瞬时的，持续时间短于一个细胞周期 早期胚胎虽然全能性的转录组已经被很好地研究，但胚胎如何利用 DUX下游的转录后和翻译调节，以建立细胞命运的这种可塑性，以及 随后什么机制限制发育潜能以确保适当的谱系确定。 最近，我们和其他人发现了p53在DUX激活中的核心作用。P53是一种主要肿瘤 抑制子和转录因子，其也激活微RNA（miRNA）34家族（a/B/c）以响应 DNA损伤。miR-34 a在许多小鼠和人类组织中表达，并且是一种被充分研究的肿瘤 抑制因子本身，抑制参与细胞生长和增殖的途径。重要的是，miR-34 a缺失 在mESCs中，赋予它们转化为全能状态的高概率，然而， 控制这种扩大的细胞命运可塑性还不清楚。我们发现mESCs中miR-34 a的缺失会导致 Dux转录物的积累，DUX 4在其3' UTR中含有预测的miR-34 a靶位点，这表明DUX 4在其3' UTR中含有预测的miR-34 a靶位点。 miR-34 a的直接调节作用。我们假设p53和miR-34 a激活和抑制Dux（和/或Dux 目标），分别形成一个反馈回路，调节全能性网络，并最终限制 体内细胞命运可塑性。揭示保护细胞命运潜力和预防 体内肿瘤发生将在癌症生物学和再生医学中产生新的见解。我独一无二 在密歇根大学亨茨曼癌症研究所的凯恩斯实验室， 犹他州。利用我们先进的DUX专业知识，结合与翻译专家的关键合作， 控制，我相信我的能力来执行这个建议中概述的实验。此外，我的 优秀的临床指导团队，从高生产力的医生和外科医生，科学家精心制作，将促进 我的临床发展，因为我试图结合联合收割机的关键见解，从分子驱动的细胞命运可塑性， 肿瘤发生转化为癌症治疗和再生医学的新靶点的开发。