Biology and Targeting of noncoding RNAs in AML

AML 中非编码 RNA 的生物学和靶向

• 批准号: 10794593
• 负责人: Ramiro Garzon
• 金额: $ 34.25万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-24 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10794593
• 关键词: APRIN gene; Acute Myelocytic Leukemia; Affect; Animals; Binding; Biogenesis; Biology; Cell Line; Cell Nucleus; Cell Proliferation; Cells; Chromosome abnormality; Classification; Clinical; Code; Complex; Cytogenetics; Cytoplasm; Data; Disease; Dose; Drug Kinetics; Ensure; Epigenetic Process; Future; Gene Expression; Gene Expression Regulation; Gene Mutation; Genes; Genetic Transcription; Genome Stability; Goals; Heterozygote; In Vitro; Leukemic Cell; Maintenance; Modeling; Molecular Abnormality; Mus; Mutate; Mutation; Myelogenous; Names; Neoplasm Metastasis; Nucleolar Proteins; Nucleotides; Outcome; Patients; Pattern; Pharmacodynamics; Plasma; Play; Pre-Clinical Model; Prognosis; Proliferating; Protein Biosynthesis; Proteins; Proteomics; RNA; RNA-Binding Proteins; Recurrence; Regulator Genes; Reporting; Ribosomes; Role; Schedule; Solid Neoplasm; Stress; Therapeutic; Toxic effect; Transcript; Transferrin; Untranslated RNA; Up-Regulation; Work; acute myeloid leukemia cell; antileukemic activity; carcinogenesis; cohort; comparative; effective therapy; experimental study; improved; in vivo; knock-down; leukemia; leukemogenesis; locked nucleic acid; loss of function; nanoparticle; novel; novel therapeutics; nucleophosmin; overexpression; patient derived xenograft model; patient subsets; pharmacodynamic model; preclinical study; prognostic significance; simulation; targeted treatment; therapeutic target; therapeutically effective; treatment strategy

The prognosis of acute myeloid leukemia (AML) is still very poor. Thus, understanding the mechanisms regulating the biology of AML is important for developing effective therapies for this disease. Non-random chromosomal abnormalities are identified in 50-55% of all AML patients. In contrast, about 45-50% of all AML cases are cytogenetically normal (CN-AML). Recent work has identified novel recurrent gene mutations in CN- AML. Among them, mutations of the nucleophosmin (NPM1) gene, represent the most common genetic alteration in CN-AML. Recently a novel class of noncoding RNAs (transcripts longer than 200 nucleotides) named long noncoding RNAs (lncRNAs) was discovered. While lncRNAs contribute to carcinogenesis in solid tumors, their role in AML has not been characterized. Our group recently identified the lncRNA HOXB-AS3 among the top up-regulated lncRNAs in NPM1 mutated (NPM1mut) CN-AML cases. We further showed that HOXB-AS3 knockdown leads to a decrease blast proliferation and colony formation in AML cell lines and primary AML patients in vitro. Silencing HOXB-AS3 in vivo using locked nucleic acid (LNA) gapmers resulted in an increased survival of treated patient derived xenograft (PDX) mice with respect to controls. Comparative proteomics identified several RNA binding protein partners of HOXB-AS3, such as EBP1, which are associated with ribosomal biogenesis. Further experiments indicated that HOXB-AS3 binds to EBP1 and regulates ribosomal biogenesis in AML by affecting the interactions between EBP1 and NPM1 complex. Altogether, our preliminary data supports our hypothesis that HOXB-AS3 plays an important role in NPM1mut AML, and blocking HOXB-AS3 may be a viable therapeutic target in NPM1mut AML. The overall goal of this is proposal is to dissect the mechanisms through which HOXB-AS3 contributes to myeloid leukemogenesis and to explore how to target therapeutically this lncRNA. We will accomplish this goal through the following Specific Aims (SA): 1) Specific Aim 1: To elucidate the mechanisms by which HOXB-AS3 promotes leukemogenesis in AML. We will perform in vitro and in vivo studies to elucidate how HOXB-AS3 modulates cell proliferation and ribosome biogenesis; 2) Specific Aim 2: To investigate in vivo the pharmacokinetic (PK), pharmacodynamics (PD) and anti-leukemic activity of a synthetic nanoparticle tagged LNA gapmer against HOXB-AS3 using PDX models of AML. In this aim we will conduct preclinical studies of synthetic Tf-NP LNA gapmer against HOXB-AS3 in PDX models of AML overexpressing HOXB-AS3 to evaluate: a) toxicity; b) plasma PK and intracellular concentrations; c) PD endpoints; d) PK/PD modeling; and e) efficacy At completion of this project, we will have an increased understanding of the role of lncRNAs in NPM1mut AML and incorporated these findings into future treatment strategies that may improve overall outcomes.

急性髓样白血病（AML）的预后仍然很差。因此，了解机制 调节AML的生物学对于开发该疾病的有效疗法很重要。非随机 在所有AML患者的50-55％中鉴定出染色体异常。相比之下，大约有45-50％的全部AML 病例在细胞遗传学上是正常的（CN-AML）。最近的工作已经确定了CN-的新型复发基因突变 AML。其中，核磷脂（NPM1）基因的突变代表了最常见的遗传 CN-AML的改变。最近，一类新的非编码RNA（转录本长于200个核苷酸） 发现了命名为长的非编码RNA（lncrnas）。而lncRNA有助于固体的癌变 肿瘤，它们在AML中的作用尚未被表征。我们的小组最近确定了lncrna hoxb-as3 在NPM1突变（NPM1MUT）CN-AML病例中，最高的上调LNCRNA。我们进一步表明 HOXB-AS3敲低导致AML细胞系中的爆炸增殖和菌落形成减少， 体外AML患者。使用锁定的核酸（LNA）间隙在体内沉默Hoxb-As3导致 相对于对照组，治疗的患者衍生异种移植（PDX）小鼠的存活率增加。比较 蛋白质组学确定了HOXB-AS3的几种RNA结合蛋白伴侣，例如EBP1，它们与之相关 与核糖体生物发生。进一步的实验表明HOXB-AS3与EBP1结合并调节 通过影响EBP1与NPM1复合物之间的相互作用，AML中的核糖体生物发生。总共，我们的 初步数据支持我们的假设，即HOXB-AS3在NPM1MUT AML中起重要作用，并且 阻断HOXB-AS3可能是NPM1MUT AML中可行的治疗靶标。这是提议的总体目标 是为了剖析HOXB-AS3有助于髓样白血病发生的机制并探索 如何以治疗性靶向此lncRNA。我们将通过以下特定目标来实现这一目标 （SA）：1）特定目标1：阐明HOXB-AS3促进白血病发生的机制 在AML。我们将进行体外和体内研究，以阐明HOXB-AS3如何调节细胞增殖 和核糖体生物发生； 2）特定目的2：在体内调查药代动力学 （PK），药效学（PD）和合成纳米颗粒标记的LNA的抗白血病活性 使用AML的PDX模型对HOXB-AS3的GAPMER。在此目标中，我们将对 AML过表达HOXB-AS3的PDX模型中的HOXB-AS3的合成TF-NP LNA Gapmer to 评估：a）毒性； b）血浆PK和细胞内浓度； c）PD端点； D）PK/PD建模；和 e）该项目完成后的功效，我们将对LNCRNA在 NPM1MUT AML并将这些发现纳入了未来的治疗策略，这些策略可能会改善整体 结果。