Hematopoiesis in germline RUNX1mutation carriers: impact of inflammation and the bone marrow niche

种系 RUNX1 突变携带者的造血作用：炎症和骨髓生态位的影响

基本信息

批准号：
10297017
负责人：
Anupriya Agarwal
金额：
$ 53.29万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10297017
关键词：
3-Dimensional Acute leukemia Affect Biogenesis Bone Marrow Bone Marrow Cells Bone Marrow Diseases Bone Marrow Transplantation CCL2 gene CRISPR/Cas technology Candidate Disease Gene Cell Differentiation process Cell Survival Cells Chronic Classification Scheme Climacteric Clinical Clonal Expansion Coculture Techniques Collection DNA Damage Data Defect Development Diagnostic Differentiation and Growth Disease Disease Progression Early identification Elements Environment Event Evolution Exposure to Extracellular Fluid Familial Platelet Disorder Family Family member First Degree Relative Gene Expression Genetic Genetic Predisposition to Disease Genetic Transcription Germ-Line Mutation Goals Growth Guidelines Hematopoiesis Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells IL8 gene Impairment In Vitro Individual Inflammation Inflammatory Institutes Knowledge Life Malignant Neoplasms Mediating Modeling Molecular Monitor Mus Mutant Strains Mice Mutate Mutation Myelogenous Pathway interactions Patient-Focused Outcomes Patients Pharmacology Phenotype Platelet aggregation Predisposition Prevention Prevention strategy Production Prognostic Marker RUNX1 gene Regulation Resources Ribosomes Risk Role Sampling Screening for cancer Signal Transduction Stress Stromal Cells Syndrome Techniques Testing Therapeutic Thrombocytopenia Time Transgenic Mice Translations Up-Regulation Variant World Health Organization Xenograft Model Xenograft procedure cytokine design early detection biomarkers erythroid differentiation experience hematopoietic differentiation improved improved outcome in vivo induced pluripotent stem cell leukemia mesenchymal stromal cell monocyte mouse model mutant mutation carrier novel novel therapeutics premalignant prevent progenitor programs response self-renewal stem therapeutic biomarker therapeutically effective tool transcription factor transcriptome transcriptomics transplant model

项目摘要

Hematopoiesis impact in germline RUNX1 mutation carriers: of inflammation and the bone marrow niche PROJECT SUMMARY/ABSTRACT Germline variants that confer risk for leukemia have been recognized increasingly with 15-20% of acute leukemia patients having at least 1 first-degree relative afflicted with leukemia. Germline variants are now included for the first time in the revised leukemia classification scheme outlined by the World Health Organization, and new clinical guidelines now include testing for inherited susceptibility as a critical element of patient diagnostics. It is devastating for these patients and their family members to live with the knowledge of having increased risk for developing cancer in their lifetimes. Thus, for these patients, early cancer detection, disease monitoring, and prevention would be life-changing, but require a comprehensive understanding of molecular changes that occur prior to overt leukemia. The long-term goal of this study is to identify the mechanisms that regulate hematopoiesis at the premalignant stage in patients with germline RUNX1 mutations [also called familial platelet disorder (FPD)]. These individuals have life-long thrombocytopenia, qualitative platelet aggregation defects, and a risk of developing a variety of hematopoietic malignancies. The acquisition of secondary mutations occurs over time in FPD patients leading to leukemia development. However, the intrinsic and/or extrinsic factors that render pre-leukemic cells vulnerable to acquire secondary mutations are unknown. To fill this knowledge gap, we performed single-cell transcriptome profiling of primary FPD bone marrow samples and identified unique transcriptional changes in FPD progenitors compared to healthy controls. Consistent these changes, FPD progenitors have impaired megakaryocytic but enhanced myeloid differentiation. Cytokine profiling of bone marrow hematopoietic and mesenchymal stromal cells (MSCs) show that there is increased cytokine production within the bone marrow, leading to chronic inflammatory stress, which may confer a growth advantage to FPD progenitors. Moreover, FPD stromal cells also show defective differentiation, clonogenic capacity, and deregulated gene expression that may further promote an inflammatory state. Thus, we hypothesize that early transcriptomic changes in FPD stem/progenitors cooperate with inflammatory microenvironmental signals to provide a growth advantage to these pre-leukemic cells and alter their differentiation. Wewill test our hypothesis using the following aims: 1)Identify how early transcriptomic changes alter hematopoiesis of FPD cells. 2) Determine the impact of inflammatory cytokines and their blockade in FPD evolution. 3) Assess the role of the FPD stromal microenvironment on growth and differentiation of FPD and healthy stem/progenitors. To achieve our goals, we have assembled several tools, including primary stem/progenitor/and stromal cells, iPSCs, xenograft and transgenic mouse models, CRISPR/Cas9 hematopoiesis targeting and editing technique, and 3D co-culture models. This project will advance our understanding of in FPD patients and will provide the molecular basis for the rational design of new therapeutics of RUNX1-mediated changes and the inflammatory microenvironment to normalize hematopoiesis delay or prevent leukemia in FPD patients.

造血作用影响在种系 RUNX1 突变携带者中：炎症和骨髓生态位项目概要/摘要人们越来越多地认识到，导致白血病风险的种系变异占 15-20% 的急性白血病风险。至少有 1 名一级亲属患有白血病的白血病患者。种系变体现在首次纳入世界卫生组织修订后的白血病分类方案组织和新的临床指南现在包括遗传易感性测试作为关键因素患者诊断。对于这些患者及其家人来说，了解这些知识是毁灭性的一生中患癌症的风险增加。因此，对于这些患者来说，早期癌症检测，疾病监测和预防将改变生活，但需要全面了解在明显的白血病之前发生的分子变化。这项研究的长期目标是确定种系 RUNX1 突变患者癌前阶段造血调节机制 [也称为家族性血小板疾病（FPD）]。这些人患有终生血小板减少症，血小板聚集缺陷，以及罹患各种造血系统恶性肿瘤的风险。此次收购随着时间的推移，FPD 患者中会发生二次突变，导致白血病的发展。然而，使白血病前期细胞容易获得二次突变的内在和/或外在因素是未知。为了填补这一知识空白，我们对原代 FPD 骨进行了单细胞转录组分析骨髓样本并鉴定出与健康人相比 FPD 祖细胞中独特的转录变化控制。与这些变化一致的是，FPD 祖细胞的巨核细胞受损，但骨髓细胞增强差异化。骨髓造血细胞和间充质基质细胞 (MSC) 的细胞因子分析显示骨髓内细胞因子的产生增加，导致慢性炎症应激，这可能赋予 FPD 祖细胞生长优势。此外，FPD 基质细胞也显示出缺陷分化、克隆形成能力和基因表达失调可能进一步促进炎症状态。因此，我们假设 FPD 干细胞/祖细胞的早期转录组变化与炎症微环境信号合作，为这些白血病前期提供生长优势细胞并改变其分化。我们将使用以下目标来检验我们的假设：1）确定多久转录组变化改变 FPD 细胞的造血作用。 2) 确定炎症细胞因子的影响以及他们对 FPD 发展的封锁。 3) 评估 FPD 基质微环境对生长和生长的作用 FPD 和健康干/祖细胞的分化。为了实现我们的目标，我们组装了多种工具，包括原代干细胞/祖细胞/和基质细胞、iPSC、异种移植和转基因小鼠模型， CRISPR/Cas9 造血作用瞄准和编辑技术和 3D 共培养模型。这个项目将加深我们对为 FPD 患者的合理设计提供分子基础 RUNX1 介导的变化和炎症微环境使造血正常化延缓或预防 FPD 患者的白血病。