microRNAs and integrative control of cardiopoiesis

microRNA 与心脏生成的综合控制

• 批准号: 8276579
• 负责人: MARK MERCOLA
• 金额: $ 50.95万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8276579
• 关键词: Address; Adult; Arizona; Biological; Biological Assay; Biological Process; Biology; Boxing; Cardiac; Cardiac Myocytes; Cardiology; Cardiovascular system; Cell Differentiation process; Cell physiology; Cells; Cellular Assay; Clinical; Collection; Data; Databases; Development; Disease; Drug Delivery Systems; Embryo; Embryonic Heart; Endoderm; Endothelial Cells; Fibroblasts; Genome; Genomic Library; Heart; Heart Diseases; Individual; Knock-out; Knowledge; Lead; Left; Libraries; Little' s Disease; Logic; Mammals; Mesoderm; MicroRNAs; Modeling; Molecular Profiling; Mus; Natural regeneration; Network-based; Oligonucleotides; Outcome; Physiology; Play; Preclinical Drug Evaluation; Prevalence; Primitive foregut structure; Process; Proteins; Proteome; Proteomics; Research; Role; Screening procedure; Signal Pathway; Signal Transduction; Signaling Protein; Small Interfering RNA; Smooth Muscle; Smooth Muscle Myocytes; Stem cells; System; Systems Analysis; Technology; Testing; Therapeutic; Validation; Vascular Endothelial Cell; Vascular Smooth Muscle; Xenopus; arm; base; cardiogenesis; cell type; follow-up; gain of function; heart cell; heart function; high throughput technology; in vivo; inhibitor/antagonist; innovation; insight; loss of function; network models; novel; progenitor; protein protein interaction; research study; small molecule; stem; stem cell differentiation

DESCRIPTION (provided by applicant): microRNAs (miRs) naturally fine-tune the responsiveness of signaling pathways, and a number of miRs are known to play important roles in heart development and disease. To date, knowledge of miRs involved in any biological process comes primarily from expression profiling. Relatively few miRs have been tested for actual regulatory roles, therefore our current knowledge of both the prevalence, importance and actual function of miRs in biological control remains rudimentary. Our data show that it is feasible to apply high throughput technology to screen stem and progenitor cell-based assays against whole genome libraries of miR oligonucleotide mimics. Using this approach, we discovered miRs that play unanticipated roles in initiating embryonic heart formation and act by distinguishing cardiac mesoderm from foregut endoderm. Here we propose to apply this same approach to identify miRs that control the next step in cardiogenesis: the differentiation and diversification cardiomyocytes, smooth muscle, and vascular endothelial cells from a common progenitor. Although the proper diversification of cardiopoietic lineages is critical to heart formation, regeneration and disease, little is known about how the process is controlled. To address this question, we will apply whole genome miR library screening, followed by an iterative process of validation of individual miRs and targets and model building to yield a refined network of signaling proteins and miRs that control cardiopoiesis. The network of miRs and target proteins will be evaluated through targeted gain and loss of function studies in embryos to visualize how miRs regulate signaling proteins in order to guide cardiopoiesis. This research will yield two outcomes: First, the functional screening and subsequent in vivo testing will reveal miRs that control cardiopoiesis and hence be important for development, disease and regeneration. Second, we will construct and test signaling networks composed of the miR targets that will offer insight into the systems-level control of cardiopoiesis. The research is innovative in that it merges state of the art functional screening of miRs with miR target identification and systems analysis in order to expose the logic underying the emergence of distinct cell types in the heart from stem cells. PUBLIC HEALTH RELEVANCE: microRNAs have emerged as key regulators of many cellular processes, including heart development and disease. However, relatively few microRNAs have been functionally tested, so appreciation of the prevalence and importance of microRNA control of cardiac biology and physiology remains fragmentary. Here we will apply high throughput drug screening technology to discover microRNAs and target proteins that control heart development, revealing how diverse types of heart cells from stem and progenitor cells.

描述（由申请人提供）：microRNAs （miRs）天然微调信号通路的反应性，并且已知许多miRs在心脏发育和疾病中发挥重要作用。迄今为止，对参与任何生物过程的miRs的了解主要来自表达谱。相对较少的miRs已经被测试为实际的调节作用，因此我们目前对miRs在生物防治中的患病率，重要性和实际功能的了解仍然是初级的。我们的数据表明，应用高通量技术筛选基于干细胞和祖细胞的miR寡核苷酸模拟物全基因组文库的检测是可行的。利用这种方法，我们发现mir在启动胚胎心脏形成中起着意想不到的作用，并通过区分心脏中胚层和前肠内胚层发挥作用。在这里，我们建议采用相同的方法来识别控制心脏发生下一步的mir：来自共同祖细胞的心肌细胞、平滑肌细胞和血管内皮细胞的分化和多样化。虽然心脏系统的适当多样化对心脏的形成、再生和疾病至关重要，但人们对这一过程是如何控制的知之甚少。为了解决这个问题，我们将应用全基因组miR文库筛选，随后是单个miR和靶点的验证和模型构建的迭代过程，以产生控制心脏生成的信号蛋白和miR的精细网络。miRs和靶蛋白的网络将通过胚胎中有针对性的功能增益和损失研究来评估，以可视化miRs如何调节信号蛋白以指导心脏生成。这项研究将产生两个结果：首先，功能筛选和随后的体内测试将揭示控制心脏生成的miRs，因此对发育、疾病和再生很重要。其次，我们将构建和测试由miR靶点组成的信号网络，这将为心脏生成的系统级控制提供见解。这项研究的创新之处在于，它将最先进的miR功能筛选与miR靶标鉴定和系统分析相结合，以揭示心脏干细胞中不同细胞类型出现的逻辑。