Biology and novel therapy of AML expressing somatic or germline mutant RUNX1

表达体细胞或种系突变体 RUNX1 的 AML 的生物学和新疗法

基本信息

批准号：
10531564
负责人：
KAPIL BHALLA
金额：
$ 48.7万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10531564
关键词：
Age Apoptosis BCL1 Oncogene BCL2 gene Back Binding Biology Bromodomain C-terminal CRISPR screen Cell Line Cell model Cells Chromatin Complex Core-Binding Factor Coupled DNA Binding DNA Polymerase II Dependence Dysmyelopoietic Syndromes Engraftment Enhancers Exhibits Exons Familial Platelet Disorder Genes Genetic Transcription Germ-Line Mutation Growth Hematopoiesis Hematopoietic stem cells Heterozygote Homologous Transplantation In Vitro Induction of Apoptosis Introns Length Libraries Malignant - descriptor Maps Mediating Messenger RNA Mus Mutation Myeloproliferative disease Network-based Oncogenes Outcome Patients Prognosis Protac Protein Isoforms Proteins RNA RUNX1 gene Reader Recurrent disease Repression Residual state Resistance Risk Somatic Mutation Time Transcriptional Activation Domain Transcriptional Regulation antagonist autosome chemotherapy chromatin protein cofactor epigenome graft vs host disease homoharringtonine improved in vivo inhibitor knock-down loss of function mutation mutant new therapeutic target novel novel therapeutics patient derived xenograft model pre-clinical preclinical efficacy recruit small hairpin RNA stem cells targeted treatment transcription factor transcriptome

项目摘要

Project Summary: RUNX1 is the DNA-binding subunit of the core binding factor (CBF) complex and a master-regulator transcription factor, which is involved in normal and malignant hematopoiesis. Somatic, heterozygous RUNX1 mutations commonly occur in Myelodysplastic Syndrome (MDS) (10%), as well as in secondary (s) or de novo AML (~10%). Germline mutations in RUNX1 cause the highly penetrant (~40%) autosomal dominant, Familial Platelet Disorder (FPD), which can evolve into myeloid malignancy (FPD-MM). Majority of mutant (mt) RUNX1 behave mostly as loss of function mutations, conferring relative therapy-resistance and poorer survival in patients with AML. Consequently, there is a strong unmet need to develop novel therapies for AML expressing somatic or germline mtRUNX1. Our preliminary studies have demonstrated for the first time that shRNA- mediated knockdown of RUNX1 (mutant and wild-type) or disruption of its binding to CBFβ induces greater lethality in AML progenitor cells (HPCs) expressing mtRUNX1 compared to wild-type (wt) RUNX1. We also found that the +24kb enhancer (eR1) within the intragenic super-enhancer (SE) of RUNX1 regulates its transcription in AML cells. The chromatin reader BET (Bromodomain Extra-terminal) protein (BETP) BRD4 promotes transcription of RUNX1 and its targets. BRD4 degradation or eviction from chromatin, or gene-editing of the +24kb RUNX1 eR1, induces lethality in AML cells. By determining and utilizing the mRNA signature from RUNX1-depleted (by shRNA) AML cells, we queried, through LINCS1000-CMap (Connectivity Mapping) analysis, for expression mimickers (EMs). We identified novel EMs that repress RUNX1 and its targets and induce significantly more apoptosis of AML cells expressing mtRUNX1 versus wtRUNX1. Therefore, the hypothesis motivating our studies is that knocking down of levels of RUNX1 and its targets will induce lethality not only in AML blasts expressing somatic mtRUNX1 but also in FPD/MM HPCs expressing germline mtRUNX1. The specific aims of studies proposed are: AIM 1: To determine impact on active enhancers, transcriptome and pre-clinical in vitro and in vivo efficacy of BETP antagonist along with its co-repression of RUNX1, BCL2 and CDK6, alone or in combination with BCL2 or CDK6 inhibitor, in AML blasts and patient- derived xenograft (PDX) models expressing somatic mutant RUNX1. Additionally, we will evaluate pre-clinical efficacy of co-targeting CRISPR-Cas9 screen-discovered top ‘druggable’ dependencies along with BETP antagonist against AML blasts expressing somatic mtRUNX1. AIM 2: To elucidate pre-clinical in vitro and in vivo efficacy of the EMs homoharringtonine (omacetaxine) or fedratinib alone and in combination with BETP antagonists against patient-derived AML blasts and PDX models expressing somatic mtRUNX1. AIM 3: To determine pre-clinical in vitro and in vivo efficacy of selected EMs that repress RUNX1 and its targets against patient-derived HPCs from FPD-MM expressing germline mtRUNX1 and other somatic co-mutations versus HPCs from RUNX1-FPD expressing only germline mtRUNX1.

项目概要： RUNX1 是核心结合因子 (CBF) 复合物的 DNA 结合亚基，也是一个主调节因子转录因子，参与正常和恶性造血作用，杂合 RUNX1。突变通常发生在骨髓增生异常综合征 (MDS) (10%) 以及继发性或新发性骨髓增生异常综合征 (MDS) 中 RUNX1 的种系突变导致 AML（~10%），导致高度外显性（~40%）常染色体显性遗传、家族性遗传。血小板疾病 (FPD)，可演变成骨髓恶性肿瘤 (FPD-MM)，大多数突变体 (mt) RUNX1。主要表现为功能丧失突变，赋予相对治疗抗性和较差的生存率经测试的 AML 患者，对于开发针对 AML 表达的新疗法存在强烈的未满足需求。我们的初步研究首次证明了shRNA-体细胞或种系mtRUNX1。介导的 RUNX1（突变型和野生型）敲低或破坏其与 CBFβ 的结合会诱导更大的与野生型 (wt) RUNX1 相比，表达 mtRUNX1 的 AML 祖细胞 (HPC) 的致死率。发现RUNX1基因内超级增强子（SE）内的+24kb增强子（eR1）调节其 AML 细胞中的转录染色质读取器 BET（溴结构域额外末端）蛋白 (BETP) BRD4 促进 RUNX1 的转录及其靶标的降解或从染色质中驱逐，或基因编辑。 +24kb RUNX1 eR1 的 mRNA 通过测定和利用 mRNA 特征诱导 AML 细胞致死。我们通过 LINCS1000-CMap（连接映射）查询了 RUNX1 耗尽（通过 shRNA）的 AML 细胞分析，我们发现了抑制 RUNX1 及其靶标的新型 EM。与 wtRUNX1 相比，表达 mtRUNX1 的 AML 细胞明显诱导更多的凋亡。激发我们研究的假设是降低 RUNX1 及其靶标的水平将导致致死不仅在表达体细胞 mtRUNX1 的 AML 母细胞中，而且在表达种系的 FPD/MM HPC 中 mtRUNX1. 拟议研究的具体目标是：目标 1：确定对活性增强剂的影响， BETP 拮抗剂的转录组和临床前体外和体内功效及其共抑制 RUNX1、BCL2 和 CDK6，单独或与 BCL2 或 CDK6 抑制剂组合，在 AML 母细胞和患者中此外，我们将评估表达体细胞突变体 RUNX1 的衍生异种移植 (PDX) 模型。共同靶向 CRISPR-Cas9 筛选发现的顶级“可成药”依赖性以及 BETP 的功效针对表达体细胞 mtRUNX1 的 AML 母细胞的拮抗剂 AIM 2：阐明体外和临床前的情况。单独使用高三尖杉酯碱（omacetaxine）或 fedratinib 以及与 BETP 联合使用的 EM 的体内疗效针对表达体细胞 mtRUNX1 的患者来源的 AML 母细胞和 PDX 模型的拮抗剂：To。确定抑制 RUNX1 及其靶标的选定 EM 的临床前体外和体内功效来自表达种系 mtRUNX1 和其他体细胞共突变的 FPD-MM 的患者来源的 HPC 来自 RUNX1-FPD 的 HPC 仅表达种系 mtRUNX1。