MICRO RNA TARGETING OF NORMAL & LEUKEMIA STEM- PROGENITOR CELLS

正常的 MICRO RNA 靶向

• 批准号: 8212935
• 负责人: CURT I CIVIN
• 金额: $ 31.13万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8212935
• 关键词: 3' Untranslated Regions; Acute Myelocytic Leukemia; Affect; Allogenic; Antigen-Presenting Cells; Apoptosis; Applications Grants; Base Pairing; Binding; Blood; Bone Marrow Transplantation; CD34 gene; CD95 Antigens; Cancer Patient; Cell physiology; Cells; Clinical; Clinical Pathways; Clinical Trials; Complication; Comprehensive Cancer Center; Data; Databases; Development; Development Plans; Erythroid; Erythropoiesis; Failure; Funding; Genes; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Human; Immunocompromised Host; In Vitro; Incubated; Individual; Informatics; Laboratories; Lead; Left; Leukemic Hematopoietic Stem Cell; Longitudinal Studies; Marrow; Messenger RNA; MicroRNAs; Modeling; Molecular; Mus; Myelogenous; Play; Predisposition; Premalignant; Process; Progress Reports; Proteins; Publishing; Recombinants; Regulation; Resistance; Resources; Role; Site; Specific qualifier value; Staging; Stem cells; T-Lymphocyte; Technology; Translational Research; Translations; Transplant Recipients; Tumor Necrosis Factor Ligand Superfamily Member 6; base; cancer stem cell; effective therapy; graft vs host disease; improved; killings; leukemia; leukemic stem cell; leukemogenesis; mRNA Expression; member; novel; prevent; progenitor; research study; scale up; stem; stem cell biology; translational approach; translational study

Project 4: MicroRNA targeting of normal and leukemia stem-progenitor cells (Civin) MicroRNAs bind to 3' untranslated region sites in target mRNAs to down-regulate translation to protein. Since microRNAs do not base-pair exactly with their target mRNAs, they can block protein translation of many mRNAs and serve as powerful switches to regulate cell functions. To extend our long-term studies on regulation of hematopoiesis and stem cell biology, we profiled microRNA expression in hematopoietic stemprogenitor cells (HSPCs). We combined this data with human HSPC mRNA expression results and microRNA-mRNA target predictions into a novel database which predicted that certain of the HSPCexpressed microRNAs (HE-microRNAs) targeted several mRNAs critical to hematopoiesis. On this informatic basis, we formulated a model for microRNA control of hematopoiesis in which many genes specifying hematopoietic differentiation are expressed by early HSPCs, but held in check by HE-microRNAs. For several target mRNAs, we then demonstrated experimentally that translation is actually decreased by microRNAs. Mir-155 potently reduced myeloid and erythroid colony formation of normal human HSPCs, and mir-16 selectively inhibited erythropoiesis. Since the cells that we studied include rare stem cells and various stages of progenitors, we propose in Aim 1, to expand our microRNA profiles of HSPCs to more highly purified subsets of primary human and mouse HSPCs and primary human acute myeloid leukemia (AMI) cells. In Aim 2, we will determine if selected individual microRNAs experimentally inhibit development of primary HSPCs, as our model predicts, and if these microRNAs affect AMI stem cells and leukemogenesis. We will determine the proteins whose synthesis is inhibited by each functionally-active microRNA and thereby the molecular mechanisms of the hematopoietic effects. At this point in our studies, at least 2 of these microRNAs, mir-16 and mir-155 appear to be new regulators of normal hematopoietic and cancer stem cells. Relevance: We suggest that a major barrier impeding cure of the many cancer patients whom we cannot cure today is our failure to effectively attack and eliminate cancer stem cells. Specifically, we believe that development of more effective therapies in leukemia hinges on fuller understanding of the regulation of rare normal and leukemic hematopoietic stem cells. The studies in this project investigate the effects of new regulatory molecules called microRNAs, which appear to play previously unexpected, potent roles in control of blood formation and leukemia. Understanding the actions of these hematopoietic-regulatory microRNAs may provide new targets, for both expansion of normal stem cells and leukemia therapy.

项目4：MicroRNA靶向正常和白血病干祖细胞（Civin） MicroRNA与靶mRNA中的3'非翻译区位点结合以下调向蛋白质的翻译。 由于microRNAs不能与它们的靶mRNA完全碱基配对，它们可以阻断蛋白质的翻译。 许多mRNA，并作为强大的开关来调节细胞功能。为了延长我们的长期研究 调节造血和干细胞生物学，我们分析了microRNA表达在造血干祖细胞， 细胞（HSPCs）。我们将该数据与人HSPC mRNA表达结果相结合， microRNA-mRNA靶向预测到一个新的数据库，预测某些HSPC表达， microRNA（HE-microRNA）靶向对造血至关重要的几种mRNA。在这个信息 在此基础上，我们建立了一个造血microRNA调控模型，其中许多基因指定 造血分化由早期HSPC表达，但由HE-microRNA控制。为 几个目标mRNA，然后我们实验证明，翻译实际上是减少了 microRNAs。Mir-155有效减少正常人HSPC的髓系和红系集落形成， miR-16选择性抑制红细胞生成。 由于我们研究的细胞包括罕见的干细胞和不同阶段的祖细胞，我们建议在 目的1，将我们的HSPCs的microRNA谱扩展到更高纯度的原代人HSPCs亚群， 小鼠HSPC和原代人急性髓性白血病（AMI）细胞。在目标2中，我们将确定 正如我们的模型预测的那样，选择的单个microRNA实验性地抑制了原代HSPC的发育， 以及这些microRNA是否影响AMI干细胞和白血病发生。我们将确定那些 合成被每种功能活性的microRNA抑制，从而抑制微RNA的分子机制。 造血作用在我们的研究中，至少有两种microRNA，mir-16和mir-155， 成为正常造血和癌症干细胞的新调节器。 相关性：我们认为，一个主要的障碍，阻碍治愈许多癌症患者，我们不能 我们今天的治疗是我们未能有效地攻击和消除癌症干细胞。具体来说，我们认为， 开发更有效的白血病治疗方法取决于更全面地了解罕见的 正常和白血病造血干细胞。该项目的研究调查了新的 被称为microRNA的调节分子，似乎在控制中发挥了以前意想不到的有效作用。 血液形成和白血病。了解这些造血调控microRNA的作用 可能为正常干细胞的扩增和白血病治疗提供新的靶点。