Cohesin Mutations in Core-Binding Factor Acute Myeloid Leukemia

核心结合因子急性髓系白血病的粘连蛋白突变

• 批准号: 10166800
• 负责人: Katelyn E Heimbruch
• 金额: $ 5.1万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10166800
• 关键词: AML1-ETO fusion protein; ATAC-seq; Acute Myelocytic Leukemia; Adult Acute Myeloblastic Leukemia; Alleles; Animal Model; Apoptosis; Automobile Driving; Basic Science; Binding; Bone Marrow Cells; Bone Marrow Transplantation; CBFB gene; CBFbeta-MYH11 fusion protein; CSPG6 gene; Cell Nucleus; Cells; ChIP-seq; Childhood; Chimeric Proteins; Chromatin; Chromosomal Rearrangement; Chromosome 16; Chromosome abnormality; Chromosomes; Clinical; Complex; Core-Binding Factor; DNA; DNA Binding; Data; Dependence; Development; Disease; Engineering; Equipment; Exhibits; Fusion Oncogene Proteins; Future; Gene Expression; Gene set enrichment analysis; Genetic; Genetic Transcription; Genomics; Goals; Hematopoiesis; In Vitro; Knowledge; Lead; Learning; Malignant Neoplasms; Mentors; Molecular; Mus; Mutation; Myelogenous; Oncogenes; Oncogenic; Outcome; Pancytopenia; Pathway interactions; Patients; Phenotype; Physicians; Play; Prognosis; RUNX1 gene; Regulation; Relapse; Research; Resources; Role; Scientist; Signs and Symptoms; Testing; Toxic effect; Training; Transplantation; Up-Regulation; XCL1 gene; base; cellular transduction; chemotherapy; cohesin; design; experimental study; genetic makeup; genetic signature; in vivo; individual patient; leukemia; leukemic transformation; leukemogenesis; member; pressure; prognostic; self-renewal; t(8; 21)(q22; q22); targeted treatment; therapeutic target; transcriptome; transcriptome sequencing; treatment strategy; virtual

Abstract Core-binding factor leukemia represents a subtype of AML and encompasses 30% of pediatric and 15% of adult AMLs. The core-binding factor complex is essential in the regulation of normal hematopoiesis and is composed of AML1 (also known as RUNX1) and CBFB. Chromosomal alterations involving the core-binding factor complex are well-known “drivers” of leukemia development and have similar clinical and prognostic implications. A t(8;21) leads to the fusion oncoprotein AML1-ETO and an inv(16) leads to CBFB-MYH11. Surprisingly, these mutations alone do not induce AML in animal models, suggesting additional “hits” are required. Mutations in one of four members of the cohesin complex (SMC1a, SMC3, RAD21, STAG1) are commonly found in patients with AML and frequently (18-20%) co-occur with AML1-ETO, but never with CBFB-MYH11. This suggests a selective pressure for the presence or absence of cohesin mutations depending upon the driver oncogene. By learning more about the genetic makeup of these leukemias and how additional mutations promote leukemogenesis, we can develop targeted therapies which will reduce both toxicity and the relapse rate compared to existing treatments. We hypothesize that cohesin mutations synergize with AML1-ETO during leukemic transformation, whereas cohesin mutations and CBFB-MYH11 form a synthetic lethal interaction. In Aim 1, in vivo transplant studies will be performed with AML1- ETO;SMC3+/+ compared to AML1-ETO;SMC3+/- while CBFB-MYH11;SMC3+/+ and CBFB-MYH11; SMC3+/- will be studied in vitro. In Aim 2, molecular studies will be performed to delve into the mechanisms driving leukemia. ChIP-sequencing will be performed to identify changes in AML1-ETO genomic occupancy upon introduction of cohesin haploinsufficiency. ATAC-seq and RNA-seq will be performed to identify changes in chromatin accessibility and transcriptome of CBFB-MYH11 due to cohesin haploinsufficiency. Our preliminary studies indicate that the loss of cohesin augments in vitro self-renewal of AML1-ETO transduced cells. Preliminary ATAC-sequencing demonstrates that several motifs implicated in myeloid development (RUNX1, GATA2, ERG, PU.1) are enriched in the AML1-ETO;SMC3+/- background. Further, RNA sequencing reveals upregulation of the MYC, Rb, and E2F oncogenic gene signatures in AML1-ETO;SMC3+/- compared to SMC3+/+. Training potential abounds for a future physician scientist within this proposal as it involves using clinical observations to generate a hypothesis, design a research plan and develop clinical correlations based upon the results of basic science research. Mentoring support will be easily accessible throughout this training, as will be all necessary equipment and resources needed to complete this project.

抽象的 核心结合因子白血病代表AML的亚型，涵盖30％的小儿和15％的成人 AMLS。核心结合因子复合物在正常造血的调节中至关重要，并且组成 AML1（也称为Runx1）和CBFB的染色体改变涉及核心结合因子 复合物是白血病发育的众所周知的“驱动力”，具有相似的临床和预后 含义。 t（8; 21）导致融合癌蛋白AML1-ETO和INV（16）导致CBFB-MYH11。 令人惊讶的是，仅这些突变不会诱导动物模型中的AML，这表明其他“命中”是 粘蛋白复合物的四个成员之一（SMC1A，SMC3，RAD21，Stag1）的突变是 通常在AML患者和经常（18-20％）与AML1-Eto同时发现，但从未发现 CBFB-MYH11。这表明根据存在或不存在粘蛋白突变的选择性压力 在驾驶员癌基因上。通过更多地了解这些白血病的遗传构成以及如何附加 突变促进了白血病，我们可以开发靶向疗法，以降低毒性和 与现有治疗相比，中继率。我们假设粘着素突变与 白血病转化期间的AML1-ETO，而粘着蛋白突变和CBFB-MYH11 形成合成的致命相互作用。在AIM 1中，将使用AML1-进行体内移植研究 与AML1-ETO相比，eTO; SMC3+/+; SMC3 +/-而CBFB-MYH11; SMC3+/+和CBFB-MYH11; SMC3 +/- Will 在体外研究。在AIM 2中，将进行分子研究以深入研究驱动机制 白血病。将进行芯片序列，以确定AML1-Eto基因组占用率的变化 引入粘蛋白单倍弥补。将执行ATAC-SEQ和RNA-SEQ，以识别 CBFB-MYH11的染色质可及性和转录组，这是由于粘着蛋白单倍不足而引起的。我们的初步 研究表明，粘着蛋白的丧失会增强AML1-Eto翻译细胞的体外自我更新。 初步的ATAC序列表明，髓样发展中实现了几个图案（Runx1， GATA2，ERG，PU.1）富含AML1-Eto; SMC3 +/-背景。此外，RNA测序揭示了 与SMC3+/+相比，AML1-Eto; SMC3 +/-中MYC，RB和E2F致癌基因特征的上调。 培训潜在的潜在为未来的物理科学家而言，因为它涉及使用临床 观察以产生假设，设计研究计划并基于 基础科学研究的结果。在整个培训中，可以轻松获得指导支持，就像 完成该项目所需的所有必要的设备和资源。