Mechanisms and therapeutic implications of human clonal hematopoiesis (CH) mutations

人类克隆造血（CH）突变的机制和治疗意义

基本信息

批准号：
10643995
负责人：
Eirini Papapetrou
金额：
$ 65.84万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10643995
关键词：
Acute Myelocytic Leukemia Advanced Malignant Neoplasm Affect Age Alleles Blood Blood Tests Bone Marrow Cancer Model Cardiovascular Diseases Cell Line Cell Survival Cells Chromatin Clinic Clonal Evolution Clonal Expansion Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Counseling DNA Methylation DNMT3a mutation Dependence Disadvantaged Dysmyelopoietic Syndromes Engineering Event Gene Mutation Genetic Models Genetic Recombination Genome Genus Mentha Goals Guidelines Hematopoiesis Hematopoietic Human In Vitro Individual Inflammatory Large-Scale Sequencing Maintenance Malignant - descriptor Malignant Neoplasms Mediating Modeling Molecular Monitor Morbidity - disease rate Mus Mutation Myeloproliferative disease Non-Hematologic Malignancy Non-Malignant Oncogenic Organism Pathology Physiological Precancerous Conditions Prevalence Process Risk Risk Estimate Role SRSF2 gene Techniques Testing Therapeutic Time Tissues Work Xenograft procedure acute myeloid leukemia cell cell immortalization cell transformation cytokine cytotoxic fitness genetic testing hematopoietic stem cell expansion in vivo induced pluripotent stem cell insight leukemia leukemogenesis mutant patient derived xenograft model premalignant stressor therapy outcome transcriptome tumorigenesis

项目摘要

PROJECT SUMMARY/ABSTRACT In recent years, large-scale sequencing revealed that the presence of clonally expanded hematopoietic stem/progenitor cells in the bone marrow and blood of healthy individuals is a widespread phenomenon in humans, generally referred to as clonal hematopoiesis (CH). Clonally expanded hematopoietic cells in many instances harbor mutations associated with myeloid malignancies and, importantly, their presence is associated with increased risk of developing myeloid malignancies (myelodysplastic syndrome, MDS and acute myeloid leukemia, AML). Leukemic progression is invariably associated with acquisition of additional mutations by the CH clone and further clonal expansions. CH is thus a premalignant condition that often constitutes the initiating event of leukemogenesis and thus offers a glimpse into the early events of malignant transformation. However, there is a relative scarcity of models to study early events of leukemogenesis, as mice do not develop CH and most human cancer models (immortalized cell lines, patient-derived xenografts) capture advanced cancers. My lab has pioneered the modeling of myeloid malignancies with human induced pluripotent stem cells (iPSCs). We recently developed a model of successive clonal evolution of AML. The overarching goal of this proposal is to investigate the molecular mechanisms underlying the oncogenic effects of CH mutations. In the first Aim we will characterize cell-intrinsic (transcriptome, chromatin accessibility, DNA methylation) and cell-extrinsic (cytokine secretion, differentiation propensity) effects of the 3 most common CH mutations – DNMT3A, TET2 and ASXL1 – using CRISPR-edited isogenic human iPSCs. In the second Aim we will explore the role of cell competition in the clonal advantage of cells harboring CH mutations before and after acquisition of additional mutations. In the third Aim, we will address the question of whether fully transformed AML cells maintain or lose dependency upon the initial CH mutation using engineered mutant alleles reversible via Cre-loxP recombination and testing the effects of correction of the CH mutation on the initiation and maintenance of AML in vitro and in vivo in xenografts. .

项目摘要/摘要近年来，大规模测序表明，克隆膨胀的造血的存在骨髓中的茎/祖细胞和健康个体的血液是一种宽度现象人类，通常称为克隆造血剂（CH）。克隆膨胀的造血细胞实例携带与髓样恶mig虫相关的突变，重要的是，它们的存在是与增加髓样恶性肿瘤的风险增加有关（骨髓增生综合征，MD和MDS 急性髓样白血病，AML）。白血病的进展总是与获取其他 CH克隆和进一步的克隆扩展的突变。因此，CH是一种经常的预先态度构成白血病的发射事件，因此可以瞥见恶性的早期事件转型。然而，模型的相对稀缺性用于研究白血病的早期事件，如小鼠不发展CH和大多数人类癌症模型（永生的细胞系，患者衍生的Xenographictics）捕获高级癌症。我的实验室通过人类诱导的多能干细胞（IPSC）。我们最近开发了一种成功的AML克隆进化模型。该提案的总体目标是研究该提议的分子机制 CH突变的致癌作用。在第一个目的中，我们将表征细胞内（转录组，染色质） 33 最常见的CH突变 - DNMT3A，TET2和ASXL1 - 使用CRISPR编辑的等源性人IPSC。在第二个目的我们将探讨细胞竞争在具有CH的细胞的克隆优势中的作用获取其他突变之前和之后的突变。在第三个目标中，我们将解决完全转化的AML细胞是否维持或失去对初始CH突变的依赖通过Cre-loxp重组可逆的工程突变等位基因，并测试CH校正的影响异种移植物中AML的主动和体内的主动和维持的突变。。