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

microRNA在调节CML干细胞自我更新和存活中的作用

• 批准号: 9207740
• 负责人: Danilo Perrotti
• 金额: $ 26.45万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9207740
• 关键词: 3' Untranslated Regions; ABL1 gene; Affect; Attention; Automobile Driving; Binding; Biological; Biological Assay; CCAAT-Enhancer-Binding Proteins; CD34 gene; Cell Proliferation; Cell Survival; Cells; Cellular biology; Chronic Myeloid Leukemia; Clinical; Dasatinib; Data; Disease; Disease Reservoirs; Dose; Drug resistance; Equilibrium; Event; Frequencies; Functional disorder; Genes; Gleevec; Goals; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic stem cells; Heterogeneous-Nuclear Ribonucleoproteins; Imatinib; Impairment; In Vitro; Individual; Knowledge; Lead; 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; Pharmacology; 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; base; bcr-abl Fusion Proteins; beta catenin; cancer stem cell; cancer survival; differential expression; hnRNP A1; in vivo; killings; leukemia; leukemic stem cell; mouse model; novel therapeutic intervention; outcome forecast; progenitor; protein E; restoration; self-renewal; stem; translational cancer research

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-ß-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-ß-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（由申请人提供）：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）治疗留下了一批对这些药物表现出先天耐药性的细胞。静态 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 HSC 且不损害正常细胞的药物。我们发表的初步数据表明，一种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 表达的改变。事实上，我们发现可能靶向 Jak2-hnRNP A1-SET/PP2A-ß-catenin 的 miR 表达发生改变。 HSC 途径，对于静止 CML HSC 的生存和自我更新至关重要，但以 BCR-ABL1 表达而非激酶依赖性方式运作。鉴于 a) BCR-ABL1 蛋白而非 mRNA 和 Jak2 表达/活性升高。在静止的 CML HSC 中；b) 白血病中的 miR 水平存在异常，并且干细胞与祖细胞中的变化；以及 c) 最近发现的 RNA 诱饵活性不仅限于miR-328 但其他 miR 可能相互作用并干扰 hnRNP 功能；推动这一提议的假设是，静止的 Ph+ HSC 表现出失调的 miR 表达，该表达依赖于 BCR-ABL1 表达而不是激酶活性，并有助于增强生存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 静态库的要求并确定治疗方案。通过鉴定和整合体外和体内分析 miR 来恢复 miR 表达的药理学恢复的相关性，miR 以典型和/或诱饵方式通过直接干扰 BCR-ABL1-Jak2 影响 CML HSC 存活和自我更新-hnRNP A1-SET-ß-连环蛋白途径因此，为了实现这些目标，我们将依靠使用高度 HSC 富集的 CD34+/CD38-(CD90+)。 BM 细胞形成 CML 患者，独特的 SCL-tTA-BCR/ABL 小鼠模型和合成 miR 或 antagomiR 治疗移植 GFP+/Luc+ CML HSC 的 NSG 小鼠，以便： 1) 识别 CML HSC 中干扰 BCR 的 miR 失调-ABL1-Jak2 途径通过其经典和/或 hnRNP A1 诱饵活性 2) 评估是否调节 miR 水平；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 miR 表达调节在根除 CML 中的治疗作用。拟议的工作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会出现新的观察结果，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。

项目成果

Cellular and Molecular Networks in Chronic Myeloid Leukemia: The Leukemic Stem, Progenitor and Stromal Cell Interplay.

• DOI: 10.2174/1389450117666160615074120
• 发表时间: 2017
• 期刊: Current drug targets
• 影响因子: 3.2
• 作者: Perrotti D;Silvestri G;Stramucci L;Yu J;Trotta R
• 通讯作者: Trotta R

BCR-ABL1 mediated miR-150 downregulation through MYC contributed to myeloid differentiation block and drug resistance in chronic myeloid leukemia.

• DOI: 10.3324/haematol.2018.193086
• 发表时间: 2018-12
• 期刊: Haematologica
• 影响因子: 10.1
• 作者: Srutova K;Curik N;Burda P;Savvulidi F;Silvestri G;Trotta R;Klamova H;Pecherkova P;Sovova Z;Koblihova J;Stopka T;Perrotti D;Polakova KM
• 通讯作者: Polakova KM

Twisting IL-1 signaling to kill CML stem cells.

扭转 IL-1 信号传导杀死 CML 干细胞。

• DOI: 10.1182/blood-2016-10-741009
• 发表时间: 2016
• 期刊: Blood
• 影响因子: 20.3
• 作者: Stramucci,Lorenzo;Perrotti,Danilo
• 通讯作者: Perrotti,Danilo

SETting OP449 into the PP2A-activating drug family.

• DOI: 10.1158/1078-0432.ccr-14-0166
• 发表时间: 2014-04-15
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Neviani P;Perrotti D
• 通讯作者: Perrotti D

Persistence of Drug-Resistant Leukemic Stem Cells and Impaired NK Cell Immunity in CML Patients Depend on MIR300 Antiproliferative and PP2A-Activating Functions.

• DOI: 10.1158/0008-5472.bcd-19-0039
• 发表时间: 2020-07
• 期刊: Blood cancer discovery
• 影响因子: 11.2
• 作者: Silvestri G;Trotta R;Stramucci L;Ellis JJ;Harb JG;Neviani P;Wang S;Eisfeld AK;Walker CJ;Zhang B;Srutova K;Gambacorti-Passerini C;Pineda G;Jamieson CHM;Stagno F;Vigneri P;Nteliopoulos G;May PC;Reid AG;Garzon R;Roy DC;Moutuou MM;Guimond M;Hokland P;Deininger MW;Fitzgerald G;Harman C;Dazzi F;Milojkovic D;Apperley JF;Marcucci G;Qi J;Polakova KM;Zou Y;Fan X;Baer MR;Calabretta B;Perrotti D
• 通讯作者: Perrotti D