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：
10308449
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/10308449
关键词：
Age Apoptosis 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 Enhancers Exhibits Exons Familial Platelet Disorder Genes Genetic Transcription Germ Lines Germ-Line Mutation Growth Hematopoiesis Hematopoietic stem cells Homologous Transplantation In Vitro Introns Length Libraries Malignant - descriptor Mediating Messenger RNA Mus Mutation Myeloproliferative disease Network-based Oncogenes Outcome Patient risk Patients Prognosis Protac Protein Isoforms Proteins RNA RUNX1 gene Reader Recurrent disease Repression Residual state Resistance Somatic Mutation Time Transcriptional Activation Domain antagonist base chemotherapy chromatin protein 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.

项目摘要： RUNX1是核心结合因子（CBF）复合物的DNA结合亚基和主调节器转录因子与正常和恶性造血有关。体细胞，杂合runx1 突变通常发生在骨髓增生综合征（MDS）（10％）中，以及次生或从头开始 AML（〜10％）。 Runx1中的种系突变导致高渗透剂（〜40％）常染色体显性剂，家族血小板疾病（FPD），可以演变为髓样恶性肿瘤（FPD-MM）。大多数突变体（MT）Runx1 行为突变的丧失，会议相对治疗 - 抗性和较差的生存率，主要是行为的行为 AML患者。因此，有很强的未满足需要开发新型AML表达的疗法体细胞或种系Mtrunx1。我们的初步研究首次证明了shrna- 介导的Runx1（突变体和野生型）的敲低或与CBFβ结合的破坏会诱导更大的与野生型（WT）Runx1相比，AML祖细胞（HPC）的致死性表达MTRUNX1。我们也是发现Runx1的基因内超增强剂（SE）中的 +24Kb增强子（ER1）调节 AML细胞中的转录。染色质读取器BET（溴化域外末端）蛋白（BETP）BRD4 促进Runx1及其目标的转录。 BRD4降解或从染色质或基因编辑中驱逐在 +24Kb runx1 er1中，诱导了AML细胞的致死性。通过确定和使用来自的mRNA签名 runx1耗尽（由shRNA）AML单元，我们通过lincs1000-cmap（连接映射）查询分析，用于表达模仿者（EMS）。我们确定了抑制Runx1及其目标的小说EMS和诱导表达MTRUNX1与WTRUNX1的AML细胞的凋亡明显更多。因此，促使我们研究的假设是，敲低Runx1及其目标的水平将引起致命性不仅在表达体细胞Mtrunx1的AML爆炸中，而且在表达种系的FPD/MM HPC中 mtrunx1。提出的研究的具体目的是：目标1：确定对主动增强子的影响， Betp拮抗剂的转录组和临床前体外和体内效率以及其共抑制在AML爆炸和患者中，单独或与Bcl2或CDK6抑制剂组合的Runx1，Bcl2和CDK6 表达躯体突变runx1的衍生异种移植（PDX）模型。此外，我们将评估临床前共同目标CRISPR-CAS9屏幕上发现顶级“可药”依赖项的功效与betp 反对表达体细胞mtrunx1的AML爆炸的对手。目标2：阐明临床前体外和 EMS同伴（omacetaxine）或Fedratinib的体内效率单独并与Betp结合反对对患者衍生的AML爆炸和表达体细胞Mtrunx1的PDX模型的拮抗剂。目标3：到确定抑制Runx1及其目标的选定EM的体外和体内效率来自FPD-MM的患者衍生的HPC表达种系Mtrunx1和其他体细胞共突变与来自Runx1-FPD的HPC仅表示种系Mtrunx1。