Investigating the Role of DNMT3A mutations in the initiation and maintenance of myeloid malignancies

研究 DNMT3A 突变在骨髓恶性肿瘤发生和维持中的作用

• 批准号: 10321210
• 负责人: Michael R Waarts
• 金额: $ 3.56万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-04 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321210
• 关键词: AML/MDS; Acute Myelocytic Leukemia; Binding Sites; CRISPR screen; CRISPR/Cas technology; Catalytic Domain; Cells; Clonal Expansion; DNA Binding; DNMT3a; DNMT3a mutation; Dependence; Development; Disease; Dysmyelopoietic Syndromes; Epigenetic Process; Epitopes; Event; FLT3 gene; Gene Frequency; Genes; Genetic; Genetic Heterogeneity; Genomics; Head; Hematopoiesis; Human; Individual; Knowledge; Laboratories; Leukemic Cell; Maintenance; Malignant - descriptor; Malignant Neoplasms; Medicine; Memorial Sloan-Kettering Cancer Center; Mentorship; Modeling; Molecular; Mutation; Myeloproliferative disease; NPM1 gene; Oncogenic; Oncology; Pathogenesis; Patients; Phenotype; Play; Precancerous Conditions; Prognosis; Proteins; Research Institute; Resources; Role; Schools; Suggestion; Tertiary Protein Structure; Therapeutic; Treatment Failure; Work; anticancer research; career development; chemotherapy; epigenome; fitness; gain of function; genetic variant; mouse model; mutant; novel; novel therapeutic intervention; premalignant; prevent; programs; recombinase; research and development; restoration; self-renewal; stem cells; therapeutic target; therapy development; treatment strategy; tumor

PROJECT SUMMARY/ABSTRACT Acute myeloid leukemia (AML) is an aggressive malignancy with a poor prognosis, owing in part to substantial intratumor genetic heterogeneity that allows specific subclones to evade even intensive chemotherapy. Genomic studies have identified the spectrum of frequent mutations in AML and suggest a model of sequential mutational acquisition. Early mutations are believed to confer a fitness advantage to a stem or progenitor cell enabling clonal expansion and later mutations within this expanding subclone may confer a proliferative advantage resulting in overt malignancy. AML can occur de novo, but may also develop in patients with myelodysplastic syndrome (MDS) or clonal hematopoiesis (CH). Across these disease states, DNMT3A mutations are critical initiating mutations, suggesting an opportunity to target the founding clone of these malignancies. However, targeting of DNMT3A mutations remains elusive due to a poor understanding of the mechanisms and specific role(s) of these mutations in disease. The objective of this proposal is to elucidate the specific oncogenic mechanisms of two frequent DNMT3A mutations and to define their functional contributions in the maintenance of CH, MDS, and AML. We hypothesize that genetic restoration of wildtype DNMT3A will block progression of DNMT3A-mutant CH/MDS/AML and thus that targeting the initiating DNMT3A mutations can provide a therapeutic option in myeloid disease. Moreover, a complete understanding of DNMT3A mutations will allow for the identification of targetable vulnerabilities conferred by these mutations. Specific Aim 1 will utilize inducible mouse models and ex vivo CRISPR screens to define the phenotypes, mechanisms, and conferred vulnerabilities of two DNMT3A mutations in distinct functional domains of the protein. Specific Aim 2 will use dual recombinase murine models capable of turning on and off DNMT3A mutations to determine the reversibility of DNMT3A-mutant CH and to evaluate the oncogenic dependency of DNMT3A mutations in MDS/AML. The implications of these studies will lead to novel therapeutic approaches for DNMT3A-mutant malignancies. This proposal will be conducted in the laboratory of Dr. Ross Levine (the Sponsor), who is the head of the Molecular Cancer Medicine program. The Levine lab is part of the Human Oncology and Pathogenesis Program at Memorial Sloan Kettering Cancer Center (MSK), a state of the art cancer research institute. Mentorship will also be provided by Dr. Kristian Helin (the Co-Sponsor), who is the head of the Center for Epigenetics at MSK. These affiliations, along with the strong scientific and non-scientific assets of the Gerstner Sloan Kettering Graduate School, will provide a rich set of collaborative, technical, and scientific resources to execute the proposed research and career development.

项目摘要/摘要 急性髓样白血病（AML）是一种侵略性恶性肿瘤，预后不良，部分原因是 肿瘤内遗传异质性允许特定的亚克隆逃避甚至强化化疗。基因组 研究已经确定了AML中频繁突变的频谱，并提出了一个顺序突变的模型 获得。据信早期突变赋予茎或祖细胞的健身优势，使克隆 扩展和以后的突变在此扩展亚克隆中可能会赋予增生优势，从而导致 明显的恶性肿瘤。 AML可能从头开始，但也可能在骨髓增生综合征患者中发展 （MDS）或克隆造血（CH）。在这些疾病状态下，DNMT3A突变是关键的启动 突变，提示有机会针对这些恶性肿瘤的创始克隆。但是，定位 DNMT3A突变仍然难以捉摸，因为对这些机制的机制和特定作用不足 疾病突变。 该提案的目的是阐明两个频繁DNMT3A的特定致癌机制 突变并定义其在CH，MD和AML维持中的功能贡献。我们假设 野生型DNMT3A的遗传恢复将阻止DNMT3A突变的CH/MDS/AML的进展，从而阻止 靶向启动DNMT3A突变可以为髓样疾病提供治疗选择。而且， 对DNMT3A突变的完全了解将允许识别可目标漏洞 由这些突变赋予。特定目标1将利用诱导鼠标模型和Ex Vivo CRISPR屏幕 定义在不同 蛋白质的功能结构域。特定的目标2将使用能够打开的双重组成酶鼠模型 并关闭DNMT3A突变，以确定DNMT3A突变物CH的可逆性并评估致癌性 MDS/AML中DNMT3A突变的依赖性。这些研究的含义将导致新的治疗性 DNMT3A突变的恶性肿瘤的方法。 该提案将在罗斯·莱文（Ross Levine）博士（赞助商）的实验室中进行，他是 分子癌医学计划。莱文实验室是人类肿瘤和发病机理计划的一部分 在最先进的癌症研究所的状态，在纪念斯隆·凯特林癌症中心（MSK）。指导将 还由MSK表观遗传学中心负责人Kristian Helin博士（共同提案国）提供。 这些隶属关系，以及Gerstner Sloan Kettering的强大科学和非科学资产 研究生院将提供一系列的合作，技术和科学资源，以执行 拟议的研究和职业发展。