Role of microRNAs in the regulation of CML stem cell survival and self renewal

microRNA在调节CML干细胞存活和自我更新中的作用

基本信息

批准号：
8429383
负责人：
Danilo Perrotti
金额：
$ 29.75万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-16 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8429383
关键词：
3&apos Untranslated Regions ABL1 gene Affect Attention Automobile Driving Binding Biological Biological Assay CCAAT-Enhancer-Binding Proteins CD34 gene Cell Proliferation Cell Survival Cells Chronic Myeloid Leukemia Dasatinib Data Disease Disease Reservoirs Dose Drug resistance Equilibrium Event Frequencies Functional disorder Genes Gleevec Goals Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells Heterogeneous-Nuclear Ribonucleoproteins Imatinib In Vitro Individual Knowledge Lead Left Leukemic Hematopoietic Stem Cell MAP Kinase Gene Maintenance Malignant - descriptor Malignant Neoplasms Mediating Messenger RNA Metabolism MicroRNAs Modeling Molecular Mus Oncogenic Pathway interactions Patients Pharmaceutical Preparations Philadelphia Phosphoric Monoester Hydrolases Phosphotransferases Play Protein Phosphatase 2A Regulatory Subunit PR53 Proteins Publishing RNA RNA-Binding Proteins Refractory Regulation Research Resistance Role Signal Transduction Stem cells Therapeutic Therapeutic Intervention Transplantation Tumor Suppressor Proteins Tyrosine Kinase Inhibitor Work anticancer research base bcr-abl Fusion Proteins cancer stem cell hnRNP A1 in vivo killings leukemia leukemic stem cell mouse model progenitor restoration self-renewal stem stem cell biology

项目摘要

DESCRIPTION (provided by applicant): Chronic myeloid leukemia (CML) is the first "clinically cured" stem cell-derived hematopoietic neoplasm; however, tyrosine kinase inhibitor (TKI) therapy leaves behind a pool of cells showing innate resistance to these drugs. These are quiescent CML stem cells (HSC) that represent an active cancer reservoir. Thus, it becomes clear that only drugs that can safely and efficiently target these HSCs without harming the normal ones have the potential to lead to disease eradication. Our published and preliminary data suggest that a likely mechanism involve an aberrant balance between kinases (i.e. BCR-ABL1 and Jak2) and phosphatases (i.e. PP2A) and this may depend on altered expression of specific microRNAs (miRs). In fact, we found altered expression of miRs that may target the Jak2-hnRNP A1-SET/PP2A-2-catenin HSC pathway, which is essential for survival and self-renewal of quiescent CML HSCs but operates in a BCR-ABL1 expression- but not kinase- dependent manner. Given that a) BCR-ABL1 protein but not mRNA and Jak2 expression/activity are elevated in quiescent CML HSCs; b) miR levels are aberrant in leukemias and change in stem vs. progenitor cells; and c) the recently discovered RNA decoy activity is not limited to miR-328 but other miRs likely interact and interfere with hnRNP function; the hypothesis driving this proposal is that quiescent Ph+ HSCs display a dysregulated miR expression which depends on BCR-ABL1 expression but not kinase activity, and contributes to enhanced survival and self-renewal of leukemic HSCs. Based on these considerations, the overall objective of this proposal is to understand the requirement of altered miR expression for the maintenance of the quiescent reservoir of CML HSCs and determine the therapeutic relevance of pharmacologic restoration of miR expression through the identification and integrated in vitro and in vivo analysis of the miRs, which in a canonical and/or decoy manner, affect CML HSC survival and self-renewal through direct interference with the BCR-ABL1-Jak2-hnRNP A1-SET-2-catenin pathway. Thus, to accomplish these goals we will rely on the use of highly HSC-enriched CD34+/CD38-(CD90+) BM cells form CML patients, the unique SCL-tTA-BCR/ABL mouse model and synthetic miRs or antagomiR to treat NSG mice transplanted with GFP+/Luc+ CML HSCs in order to: 1) Identify miRs dysregulated in CML HSCs that interfere with the BCR-ABL1-Jak2 pathway through their canonical and/or hnRNP A1 decoy activities; 2) Assess whether modulation of miR levels impairs in vitro and in vivo survival and self-renewal of CML HSCs; and 3) Determine the therapeutic role of modulation of miR expression in eradication of CML by using 2-O-MePS miRs and antagomiRs. We are confident that the successful completion of the proposed work will advance our knowledge of leukemic HSC biology and, based on the discoveries we made in the past few years, it is safe to predict that new observations will be made in this field and that some of them will reveal new strategies for therapeutic intervention. Hence, the strong importance and high relevance of this work for basic and translational cancer research.

描述（申请人提供）：慢性粒细胞白血病（CML）是第一种“临床治愈”的干细胞源性造血肿瘤；然而，酪氨酸激酶抑制剂（TKI）疗法留下了一批对这些药物表现出先天抵抗力的细胞。这些是静态的 CML 干细胞 (HSC)，代表活跃的癌症储存库。因此，很明显，只有能够安全有效地靶向这些造血干细胞而不伤害正常造血干细胞的药物才有可能消除疾病。我们发表的初步数据表明，可能的机制涉及激酶（即 BCR-ABL1 和 Jak2）和磷酸酶（即 PP2A）之间的异常平衡，这可能取决于特定 microRNA (miR) 表达的改变。事实上，我们发现 miR 的表达发生改变，可能靶向 Jak2-hnRNP A1-SET/PP2A-2-连环蛋白 HSC 通路，该通路对于静止 CML HSC 的生存和自我更新至关重要，但在 BCR-ABL1 表达中起作用 -但不是激酶依赖性方式。鉴于 a) 在静态 CML HSC 中，BCR-ABL1 蛋白升高，但 mRNA 和 Jak2 表达/活性升高； b) 白血病中的 miR 水平异常，并且干细胞与祖细胞中的变化； c) 最近发现的 RNA 诱饵活性不仅限于 miR-328，其他 miR 也可能与 hnRNP 功能相互作用并干扰；推动这一提议的假设是，静态 Ph+ HSC 表现出 miR 表达失调，该表达依赖于 BCR-ABL1 表达而不是激酶活性，并有助于增强白血病 HSC 的存活和自我更新。基于这些考虑，本提案的总体目标是了解改变 miR 表达对于维持 CML HSC 静态储存库的要求，并通过体外和体内鉴定和整合来确定 miR 表达药理学恢复的治疗相关性。 miR 的体内分析，通过直接干扰 BCR-ABL1-Jak2-hnRNP，以典型和/或诱饵方式影响 CML HSC 存活和自我更新A1-SET-2-连环蛋白途径。因此，为了实现这些目标，我们将依靠使用来自 CML 患者的高度 HSC 富集的 CD34+/CD38-(CD90+) BM 细胞、独特的 SCL-tTA-BCR/ABL 小鼠模型和合成 miR 或 antagomiR 来治疗 NSG 小鼠移植 GFP+/Luc+ CML HSC 以便： 1) 识别 CML HSC 中失调的 miR，这些 miR 会干扰BCR-ABL1-Jak2 途径通过其经典和/或 hnRNP A1 诱饵活性； 2) 评估miR水平的调节是否损害CML HSC的体外和体内存活和自我更新； 3) 使用 2-O-MePS miR 和 antagomiR 确定调节 miR 表达在根除 CML 中的治疗作用。我们相信，拟议工作的成功完成将增进我们对白血病 HSC 生物学的了解，并且根据我们过去几年的发现，可以安全地预测该领域将出现新的观察结果，并且一些其中将揭示治疗干预的新策略。因此，这项工作对于基础和转化癌症研究具有很强的重要性和高度相关性。