The role of the cohesin complex in hematopoietic transformation and leukemia maintenance

粘连蛋白复合物在造血转化和白血病维持中的作用

基本信息

批准号：
10780841
负责人：
AARON D VINY
金额：
$ 48.74万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
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：
2023-09-19 至 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10780841
关键词：
3-Dimensional AML/MDS Acute Myelocytic Leukemia Acute Promyelocytic Leukemia Affect Alleles Biological Assay Biological Markers Blood Cells Bone Marrow Bone Marrow Stem Cell Cell Lineage Cells Charge Chromatin Chromatin Loop Chromatin Structure Complex Credentialing Cytarabine Cytotoxic agent Dependence Disease remission Doxorubicin Dysmyelopoietic Syndromes Enhancers Epigenetic Process Equilibrium Euchromatin Event FLT3 gene FLT3 inhibition Functional disorder Gene Expression Genetic Genetic Transcription Genomics Hematopoiesis Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells Heterochromatin Hi-C Homeostasis Human Impairment In Vitro Leukemic Hematopoietic Stem Cell Maintenance Malignant - descriptor Malignant Neoplasms Maps Mediating Mission Modeling Molecular Profiling Mus Mutagenesis Mutate Mutation Myelogenous Myeloproliferative disease Neighborhoods Output Patients Phenocopy Phenotype Play Population Prediction of Response to Therapy Production Recurrence Regimen Regulation Repression Resources Role Sampling Secondary acute myeloid leukemia Serial Passage Strategic Planning Structure Survival Rate System Techniques Technology Therapeutic Tissue-Specific Gene Expression Transcriptional Regulation Treatment Protocols Tretinoin Validation acute myeloid leukemia cell biobank cancer subtypes chemotherapy chromatin immunoprecipitation chromatin remodeling cohesin driver mutation epigenomics gene regulatory network hematopoietic differentiation hematopoietic stem cell differentiation hematopoietic stem cell self-renewal hip replacement arthroplasty in vivo inhibitor innovative technologies insight leukemia leukemic transformation leukemogenesis loss of function member mouse model multiple omics mutant mutant mouse model mutational status new therapeutic target novel patient derived xenograft model prevent progenitor programs recombinase restoration self-renewal stem stem cell proliferation stem cell self renewal targeted treatment therapy resistant transcription factor transcriptome sequencing treatment response tumorigenesis

项目摘要

PROJECT SUMMARY In this proposal, we will define the effects of epigenetic dysregulation via Stag2 loss on three-dimensional chromatin structure and gene expression. We will define key lineage-specific chromatin features and validate their hierarchy in differentiation in contrast to stem cell self-renewal. In Aim 1, we will perform low-input Hi-ChIP assay on Ctcf (loop/TAD-boundary), Rad21 (pan-cohesin loop), ChIP assay for H3K27Ac (enhancer) and RNA- seq (gene expression) on phenotypically defined mouse and human stem and progenitors to define population- specific looping events. Using our novel ‘on-off-on’ dual recombinase (Stag2GL) allele, we will first determine if the restoration of Stag2 can reverse the effects on self-renewal/differentiation and define chromatin loops 1) perturbed during Stag2 loss and 2) restored after Stag2 reversion. Locus-specific functional assays will determine the key events required for self-renewal through in vitro studies to phenocopy in ‘Stag2-on' or prevent reversion in ‘Stag2-off’. In Aim 2, using our low cell input Hi-C and Hi-ChIP techniques to assess chromatin loops alterations identified in our fully penetrant Stag2/Flt3 co-mutant AML models in combination with the Stag2GL allele. Using a multi-omics approach, we will define the chromatin landscape of Flt3 with and without the antecedent Stag2 mutation. We have created a well-annotated MDS and AML patient biorepository, as well as a biorepository of healthy bone marrow from hip arthroplasty patients. Using these primary patient samples, we will map chromatin loops and transcriptional output in stem and progenitor populations and intersect lineage- specific interactions in human and mouse. Functional validation of chromatin loops lost or gained in the co- mutant models will be assessed in vitro, prioritizing loci identified in primary AML patient samples. Stag2 reversion in this system will definitively determine whether Stag2 is required for the maintenance of AML and capability for chromatin reversibility, hereby representing a genetic dependency in Stag2-mutant AML. Finally, these mouse models and serially passaging patient derived xenograft resources will constitute the systems to assess the effects of Stag2 loss on therapeutic response to standard chemotherapeutic regimens (cytarabine and doxorubicin) as well as to emerging targeted therapies such as FLT3 inhibition in FLT3-ITD mutant AML (gilteritinib). Taken together, the results of this proposal will elucidate the pathophysiological mechanism of Stag2, leading to a deeper understanding of 1) epigenetic mechanisms of chromatin remodeling on transcriptional regulation, epigenetic plasticity, 2) the contributions of Stag2 loss towards tumorigenesis in MDS and secondary AML and 3) identification of key novel therapeutic targets.

项目摘要在此提案中，我们将通过Stag2损失对表观遗传失调的影响对三维染色质结构和基因表达。我们将定义关键谱系特异性染色质特征并验证与干细胞自我更新相比，它们在分化方面的层次结构。在AIM 1中，我们将执行低输入Hi-Chip 对CTCF（环/t-boundary），Rad21（泛粘蛋白环），H3K27AC（增强剂）和RNA-的测定法测定在表型定义的小鼠和人类茎以及祖细胞上的Seq（基因表达），以定义种群特定的循环事件。使用我们的小说“ On-On-On-On-On-On-On-On-On-On-On Dual重组酶（Stag2GL）等位基因，我们将首先确定是否是否 stag2的恢复可以扭转对自我更新/分化的影响，并定义染色质环1）在stag2丢失期间受到干扰和2）逆转后恢复。特定于基因座的功能测定将通过体外研究确定自我更新所需的关键事件 “ stag2-off”中的归还。在AIM 2中，使用我们的低细胞输入HI-C和HI-CHIP技术来评估染色质环路在我们完全渗透的Stag2/flt3共突变AML模型中确定的变化与Stag2Gl结合使用多词方法，我们将在有和没有的情况下定义FLT3的染色质景观先决的stag2突变。我们创建了一个通知良好的MDS和AML患者生物库，以及来自髋关节置换术患者的健康骨髓的生物座。使用这些主要患者样本，我们将绘制茎和祖先种群中的染色质环和转录输出，并与谱系相交人和小鼠中的特定相互作用。在共同体中丢失或获得的染色质环的功能验证将在体外评估突变模型，并确定本地式AML患者样品中的优先级。 Stag2 该系统中的归还将确定确定是否需要Stag2维护AML和染色质可逆性的能力，特约代表Stag2突变AML中的遗传依赖性。最后，这些鼠标模型和连续传递的患者派生的定义资源将构成系统评估STAG2损失对治疗对标准化学治疗方案的反应的影响（Cytarabine 和阿霉素）以及新兴的靶向疗法，例如FLT3-ITD突变体AML中的FLT3抑制作用（Gilteritinib）。综上所述，该提案的结果将阐明 stag2，导致对1）染色质重塑的表观遗传机理的深入了解转录调节，表观可塑性，2）Stag2损失对MDS的肿瘤发生的贡献和次级AML和3）鉴定关键的新型治疗靶标。