MicroRNA Function in Human Embryonic Stem Cells

MicroRNA 在人类胚胎干细胞中的功能

• 批准号: 7908076
• 负责人: Hannele RUOHOLA-BAKER
• 金额: $ 12.45万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7908076
• 关键词: Adult; Binding; Biological Assay; Cell physiology; Cells; Dicer Enzyme; Drosophila genus; Functional RNA; Future; Gene Targeting; Genes; Genetic Translation; Goals; Informatics; Mediating; Messenger RNA; MicroRNAs; Mus; Phenotype; Plants; Process; RNA Interference; Regenerative Medicine; Regulation; Research; Role; Stem cells; Testing; Tissues; Transplantation; aptamer; base; embryonic stem cell; gain of function; genome-wide; human DICER1 protein; human embryonic stem cell; loss of function; public health relevance; research study; self-renewal; small molecule; tumor

DESCRIPTION (provided by applicant): The long-term goal of this research is to gain a better understanding and control over human embryonic stem cells (hESCs) by identifying and manipulating the key microRNAs required for hESC self-renewal and differentiation. MicroRNAs, small (21-23nt) non-coding RNA molecules that can regulate mRNA translation and stability by binding to the target mRNAs with incomplete complementarity, are required for normal stem cell function in mouse, Drosophila and plants. We have previously shown that microRNAs are required for germline stem cell (GSC) self-renewing divisions in Drosophila. Our preliminary results suggest that microRNAs are also critical for stem cell self-renewing division and differentiation in human ESCs, and the goal of this proposal is to identify the critical microRNAs and their mRNA targets in these cells. Towards this goal, we will analyze the phenotypes observed in hESCs lacking mature microRNAs due to gene knockdown (KD) of microRNA processing enzymes, Dicer or Drosha. We will identify up-regulated mRNAs in these KD-lines by genome wide mRNA profiling and determine whether they are responsible for the Dicer or Drosha phenotypes through loss-of-function (LOF), gain-of-function (GOF) and rescue experiments (Aim1). To identify the critical microRNAs that mediate this regulation we will carry out genome wide QPCR profiling of hESC microRNAs. The function of the hESC enriched microRNAs will be analyzed by LOF, GOF and rescue experiments (Aim2). The two aims will connect: The critical microRNAs should have the capacity to rescue dicer KD phenotypes (Aim 2.2) and may be sufficient to down-regulate the target genes identified in Aim 1.2 (Aim 2.3). This study should result in findings that allow better manipulation and more efficient differentiation of hESC which is of particular importance for regenerative medicine. PUBLIC HEALTH RELEVANCE: This study will be the first analysis of the functions of mature miRNAs in hESCs. Identification of the miRNA function in hESCs holds the future for better control and manipulation of the hESCs in regenerative medicine. This is important due to the troubling fact that embryonic stem cells tend to form tumors when transplanted into adult tissue. microRNAs carry hope as molecules which can control stem cell self-renewing division.

描述（由申请人提供）：这项研究的长期目标是通过识别和操纵hESC自我更新和分化所需的关键microRNA来更好地理解和控制人类胚胎干细胞（HESC）。 microRNA，小（21-23NT）非编码RNA分子，可以通过与靶mRNA结合具有不完全互补性来调节mRNA平移和稳定性，这对于小鼠，果蝇和植物中的正常干细胞功能是必需的。我们以前已经表明，果蝇中种系干细胞（GSC）自我更新师必需的microRNA。我们的初步结果表明，microRNA对于人类ESC的干细胞自我更新分裂和分化也至关重要，该建议的目的是确定这些细胞中关键的microRNA及其mRNA靶标。为了实现这一目标，我们将分析由于microRNA加工酶，迪切尔（Dicer）或drosha的基因敲低（KD）而缺乏成熟microRNA的hESC中观察到的表型。我们将通过基因组广泛的mRNA分析来鉴定这些KD线中的上调mRNA，并确定它们是否通过功能丧失（LOF），功能获得（GOF）和救援实验负责DICER或DROSHA表型（AIM1）。为了确定介导该调节的关键microRNA，我们将对hESC microRNA进行基因组范围的QPCR分析。 LOF，GOF和救援实验将分析HESC富集的microRNA的功能（AIM2）。这两个目标将连接：关键的microRNA应具有挽救DICER KD表型的能力（AIM 2.2），并且可能足以下调AIM 1.2中确定的目标基因（AIM 2.3）。这项研究应导致发现可以更好地操纵和更有效地分化hESC，这对于再生医学尤为重要。公共卫生相关性：这项研究将是对成熟miRNA在hESC中的功能的首次分析。 hESC中miRNA功能的识别具有更好地控制和操纵再生医学中的hESC的未来。这很重要，因为令人不安的事实是，将胚胎干细胞移植到成人组织中时倾向于形成肿瘤。 MicroRNA具有希望，作为可以控制干细胞自我更新分裂的分子。