Biology and Targeting of noncoding RNAs in AML

AML 中非编码 RNA 的生物学和靶向

基本信息

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
项目类别：
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

项目摘要

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.

急性髓系白血病（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非常重要。非随机 50-55% 的 AML 患者存在染色体异常。相比之下，大约 45-50% 的 AML 病例细胞遗传学正常（CN-AML）。最近的工作发现了 CN-中新的复发基因突变反洗钱。其中，核磷蛋白（NPM1）基因的突变代表了最常见的基因突变。 CN-AML 的改变。最近出现了一类新型非编码 RNA（转录本长度超过 200 个核苷酸）发现了长非编码RNA（lncRNA）。虽然lncRNA在固体中有助于致癌肿瘤，其在 AML 中的作用尚未得到表征。我们的团队最近鉴定出了lncRNA HOXB-AS3 NPM1 突变 (NPM1mut) CN-AML 病例中上调最多的 lncRNA 之一。我们进一步表明 HOXB-AS3 敲低导致 AML 细胞系中母细胞增殖和集落形成减少体外原发性 AML 患者。使用锁核酸 (LNA) 间隙体在体内沉默 HOXB-AS3 导致与对照组相比，经过治疗的患者来源的异种移植（PDX）小鼠的存活率有所提高。比较蛋白质组学鉴定了 HOXB-AS3 的几种 RNA 结合蛋白伴侣，例如 EBP1，它们与与核糖体生物发生。进一步的实验表明 HOXB-AS3 与 EBP1 结合并调节通过影响 EBP1 和 NPM1 复合物之间的相互作用来调节 AML 中的核糖体生物合成。总而言之，我们的初步数据支持我们的假设，即 HOXB-AS3 在 NPM1mut AML 中发挥重要作用，并且阻断 HOXB-AS3 可能是 NPM1mut AML 的可行治疗靶点。本次提案的总体目标是目的是剖析 HOXB-AS3 促进髓系白血病发生的机制并探索如何针对该 lncRNA 进行治疗。我们将通过以下具体目标来实现这一目标 (SA): 1) 具体目标1：阐明HOXB-AS3促进白血病发生的机制在反洗钱中。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) 具体目标2：研究体内药代动力学标记 LNA 的合成纳米颗粒的 (PK)、药效学 (PD) 和抗白血病活性使用 AML 的 PDX 模型针对 HOXB-AS3 的gapmer。为了这个目标，我们将进行临床前研究在 AML 过表达 HOXB-AS3 的 PDX 模型中合成 Tf-NP LNA gapmer 对抗 HOXB-AS3 评估： a) 毒性； b) 血浆药代动力学和细胞内浓度； c) PD终点； d) PK/PD建模；和 e) 功效本项目完成后，我们将对 lncRNA 在以下方面的作用有更深入的了解： NPM1mut AML 并将这些发现纳入未来的治疗策略中，可能会整体改善结果。