MicroRNA Regulation of Stem Cell Self-renewal

MicroRNA 调控干细胞自我更新

• 批准号: 8208209
• 负责人: Shangqin Guo
• 金额: $ 15.37万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-24 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8208209
• 关键词: 3' Untranslated Regions; Adult; Biochemical; Biogenesis; Bone Marrow; Candidate Disease Gene; Cell physiology; Cells; Collaborations; Colony-Forming Units Assay; Computer Analysis; Development; Dicer Enzyme; Genes; Genetic Screening; Genetically Engineered Mouse; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic stem cells; In Vitro; Injury; Knockout Mice; Laboratories; Malignant - descriptor; Messenger RNA; MicroRNAs; Molecular; Molecular Profiling; Molecular Target; Mouse Strains; Mus; Natural regeneration; Pathway interactions; Phenotype; Plant Roots; Population; Principal Investigator; Process; Proteins; Recording of previous events; Refractory; Regulation; Research; Role; Scanning; Staging; Stem cells; Surveys; System; Technical Expertise; Therapeutic; Time; Tissues; Transplantation; base; cancer stem cell; comparative genomics; embryonic stem cell; in vitro Assay; in vivo; leukemia; leukemic stem cell; mRNA Expression; mouse model; programs; self-renewal; stem cell biology; success; tool

Cell intrinsic mechanisms exist to regulate hematopoietic stem cell (HSC) self-renewal. Self-renewal ability confined to the normal stem cells is essential for maintaining tissue integrity and is responsible for regeneration after injury. When acquired by aberrant cells, however, it could produce a self-renewing cancer stem cell that is refractory to treatments. Understanding the mechanisms of self-renewal is important for therapeutic expansion of HSCs as well as for eradicating the malignant roots of blood cancers. Contrasting to mRNA expression, microRNA expression faithfully marks developmental history and/or stage. HSCs develop through a well characterized step-wise differentiation process with a gradual loss of the self- renewing capacity. Disruption of the microRNA biogenesis key enzyme, Dicer, completely ablated stem cell activity. We thus hypothesized that specific microRNAs are required in regulating these key steps and the expression of some microRNAs may correlate with the ability to self-renew. Two complementary approaches were used to identify such specific microRNAs. By scanning the 3'UTR regions of genes known to be important for hematopoiesis, we identified that miR-150 exerted its function by targeting c-Myb. Additionally, microRNA expression profiling was performed using primary stem cells and their non-renewing immediate descendant cells. Based on the microRNA expression profile, we identified the miR-99b-let-7e-miR-125a cluster which dramatically expanded the hematopoietic compartment after long term transplantation in vivo. propose to gain detailed understanding of these microRNA species in regulating HSC self-renewal through two specific aims. 1. Determine the role of miR-150 in HSC function using miR-150 knockout mice. 2. Define the role of the miR-99b-let-7e-125a cluster in enhancing HSC self-renewal and assess its candidate molecular targets. These aims will be achieved through the combinational use of genetically engineered mouse strains and the appropriate in vivo and in vitro assays readily available in Dr. Scadden's laboratory. RELEVANCE (Seeinstructions): The proposed study promises the discovery of microRNA regulators for HSC self-renewal. Such microRNAs will provide new paradigms for HSC regulation. They will also serve as research tools to uncover additional molecular pathways and networks that confer the self-renewal phenotype, not only to normal HSCs, but also to leukemia stem cells.

造血干细胞（HSC）的自我更新是由细胞内在机制调控的。自我更新能力 局限于正常干细胞是维持组织完整性所必需的， 损伤后的再生然而，当被异常细胞获得时，它可以产生自我更新的癌症 对治疗无效干细胞。了解自我更新的机制对于 HSC的治疗性扩增以及用于根除血癌的恶性根源。 与mRNA表达相比，microRNA表达忠实地标记了发育历史和/或阶段。 HSC通过充分表征的逐步分化过程发育，随着自分化能力的逐渐丧失， 更新能力。microRNA生物发生关键酶Dicer的破坏完全消融了干细胞 活动因此，我们假设特定的microRNA是调节这些关键步骤所必需的， 一些microRNA的表达可能与自我更新的能力相关。两种互补的方法 来鉴定这些特定的microRNA。通过扫描已知的基因的3 'UTR区域， 对于造血重要，我们鉴定了miR-150通过靶向c-Myb发挥其功能。此外，本发明的目的是， 使用原代干细胞和它们的非更新性即刻细胞进行microRNA表达谱分析。 后代细胞基于microRNA表达谱，我们鉴定了miR-99 b-let-7 e-miR-125 a 簇，其在体内长期移植后显著扩大造血隔室。 我建议通过以下途径详细了解这些microRNA种类在调节HSC自我更新中的作用： 两个具体目标。1.使用miR-150敲除小鼠确定miR-150在HSC功能中的作用。2.定义 miR-99 b-let-7 e-125 a簇在增强HSC自我更新中的作用，并评估其候选者 分子靶点这些目标将通过结合使用基因工程 小鼠品系和适当的体内和体外试验，可在Scadden博士的实验室中获得。 相关性（参见说明）： 这项研究有望发现HSC自我更新的microRNA调节剂。这些microRNA 将为HSC监管提供新的范例。它们还将作为研究工具， 这些分子通路和网络不仅赋予正常的HSC自我更新表型，而且还 到白血病干细胞。