Cellular and molecular basis of microRNA-29a Induced Acute Myeloid Leukemia

microRNA-29a 诱导的急性髓系白血病的细胞和分子基础

• 批准号: 8580098
• 负责人: CHRISTOPHER Y PARK
• 金额: $ 36.69万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-10 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8580098
• 关键词: Acute Myelocytic Leukemia; Apoptosis; Appearance; Biological Assay; Biological Models; Blood Cells; Cell Cycle; Cell physiology; Cell surface; Cells; Ceramides; Clinical; Commit; Data; Development; Disease; Ectopic Expression; Epigenetic Process; Event; Exhibits; Gene Targeting; Genes; Genetic; Gold; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Human; Lesion; MLL-AF9; Maintenance; Malignant Neoplasms; Mediator of activation protein; MicroRNAs; Modeling; Molecular; Mus; Mutate; Mutation; Myelogenous; Myeloid Leukemia; Myeloproliferative disease; Outcome; Pathogenesis; Phase; Phenotype; Population; Process; RUNX1 gene; Regulation; Relative (related person); Role; Sampling; Somatic Mutation; Staging; Stem cells; Study models; Testing; Transplantation; Work; base; experience; genetically modified cells; human disease; in vivo; insight; leukemia; leukemic stem cell; leukemogenesis; mouse model; new therapeutic target; progenitor; public health relevance; self-renewal; stem; stem cell population; therapeutic target; transcriptome sequencing

DESCRIPTION (provided by applicant): Cellular and molecular basis of microRNA-29a induced acute myeloid leukemia. Acute myeloid leukemia (AML) arises from the accumulation of genetic and/or epigenetic changes in immature hematopoietic cells. While leukemogenesis involves multiple steps including initiation, progression, transformation, and maintenance, studying these processes in human disease presents many practical challenges; however, well-characterized mouse models of AML afford the opportunity to study each of them due to the relative ease of genetically modifying cells and testing cellular function in a variety of in vitroand in vivo assays. While AML arises from immature hematopoietic cells, the exact cell that initiates disease or ultimately transforms into the leukemia stem cell (LSC) is unclear. We previously showed that microRNA-29a (miR-29a) is highly expressed in human HSC and AML LSC, and that ectopic expression of miR-29a in mouse BM cells is sufficient to induce a myeloproliferative neoplasm (MPN)-like disease that progresses to AML. We characterized miR-29a induced disease and showed that self-renewing committed progenitors arise prior to the development of AML and that miR-29a induced AMLs contain LSCs that can be prospectively isolated based on the expression of specific cell surface markers. Although the precise role of the self-renewing progenitors has not been defined, their appearance raises the intriguing possibility that aberrant acquisition of self-renewal by committed progenitors may be an early event in AML pathogenesis. Overall, these data demonstrate that the miR-29a induced AML model is a robust and experimentally tractable model for studying AML pathogenesis. We propose to characterize miR-29a's roles during AML development and to elucidate the molecular basis of miR-29a's regulation of self-renewal in LSCs and aberrantly self-renewing progenitors. We will determine if miR-29a is required for leukemogenesis utilizing our newly developed miR-29a deficient mouse model as well as various mouse models of AML. We will identify the cell population/s that become transformed during miR- 29a driven leukemogenesis, and we will also test the contribution of two miR-29a targets, Dnmt3a and Smpd3 (genes also recurrently mutated in human AML) to miR-29a induced phenotypes. To identify potential genes or mutations that may cooperate with miR-29a to induce leukemia, we will perform RNA-Seq on LSCs and aberrantly self-renewing progenitors. Finally, we will leverage our experience working with the miR-29a induced AML model as well as with primary human AML samples to evaluate the role of miR-29a in LSC function in the serial transplantation setting, the gold-standard assay for self-renewal. Ultimately, we expect these studies to generate new insights into the role of miR-29a in AML, define the molecular mechanisms of miR-29a's actions in pre-leukemic intermediates (including the molecular basis for self-renewal), and determine whether miR-29a or its downstream mediators may serve as potential therapeutic targets in AML.

描述（由申请人提供）：microRNA-29A的细胞和分子基础诱导急性髓样白血病。急性髓细胞性白血病（AML）源于未成熟造血细胞中遗传和/或表观遗传变化的积累。虽然白血病涉及多个步骤，包括开始，进展，转型和维持，但研究人类疾病中的这些过程带来了许多实际挑战。然而，由于遗传修饰细胞的相对易于易于易于易于，并且在各种体外体内测定中测试细胞功能，因此有机会研究它们的小鼠模型。尽管AML来自未成熟的造血细胞，但尚不清楚造成疾病或最终转化为白血病干细胞（LSC）的确切细胞尚不清楚。我们先前表明，microRNA-29a（miR-29a）在人HSC和AML LSC中高度表达，并且在小鼠BM细胞中miR-29a的异位表达足以诱导发展为AML的脊髓增生性肿瘤（MPN）类似于AML。我们表征了miR-29a诱导的疾病，并表明在AML发展之前出现了自我更新犯的祖细胞，并且miR-29a诱导的AML含有LSC，可以根据特定细胞表面标记的表达来前瞻性地分离出LSC。尽管尚未定义自我更新祖细胞的确切作用，但它们的外观增加了一种有趣的可能性，即犯罪祖细胞对自我更新的异常获得可能是AML发病机理中的早期事件。总体而言，这些数据表明，miR-29a诱导的AML模型是用于研究AML发病机理的强大且具有实验性探索模型。我们建议在AML发育过程中表征miR-29a的作用，并阐明miR-29a在LSC中调节自我更新和异常更新的祖细胞的分子基础。我们将利用新开发的miR-29a缺陷小鼠模型以及AML的各种小鼠模型来确定白血病发生是否需要miR-29a。我们将确定在miR-29a驱动的白血病中转化的细胞群，我们还将测试两个miR-29a靶标的DNMT3A和SMPD3（在人AML中也反复突变）对miR-29a诱导的表型的贡献。为了鉴定可能与miR-29a合作以诱导白血病的潜在基因或突变，我们将对LSC和异常自我更新的祖细胞进行RNA-Seq。最后，我们将利用与miR-29a诱导的AML模型以及原代人AML样品一起工作的经验，以评估miR-29a在LSC功能在串行移植设置中的作用，这是自我更新的金标准测定法。最终，我们期望这些研究能够产生对miR-29a在AML中的作用的新见解，它定义了miR-29a在美氏学前中间体（包括自我恢复的分子基础）中的作用的分子机制，并确定miR-29a或其下游介体是否可以用作AML中的潜在治疗靶标。