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.

项目概要/摘要 急性髓系白血病 (AML) 是一种侵袭性恶性肿瘤，预后不良，部分原因是大量的 肿瘤内遗传异质性使得特定亚克隆甚至可以逃避强化化疗。基因组 研究已经确定了 AML 中常见突变的范围，并提出了一种连续突变模型 获得。早期突变被认为赋予干细胞或祖细胞适应性优势，从而实现克隆 该扩展亚克隆内的扩展和后来的突变可能会赋予增殖优势，从而导致 明显的恶性肿瘤。 AML 可以从头发生，但也可能发生在骨髓增生异常综合征患者中 (MDS) 或克隆造血 (CH)。在这些疾病状态中，DNMT3A 突变是关键的起始 突变，表明有机会针对这些恶性肿瘤的创始克隆。然而，针对 由于对这些突变的机制和具体作用了解甚少，DNMT3A 突变仍然难以捉摸。 疾病的突变。 该提案的目的是阐明两种常见 DNMT3A 的具体致癌机制 突变并确定其在维持 CH、MDS 和 AML 中的功能贡献。我们假设 野生型 DNMT3A 的基因恢复将阻止 DNMT3A 突变体 CH/MDS/AML 的进展，从而 靶向 DNMT3A 起始突变可以为骨髓疾病提供一种治疗选择。此外，一个 完整了解 DNMT3A 突变将有助于识别可针对的漏洞 这些突变所赋予的。具体目标 1 将利用诱导型小鼠模型和离体 CRISPR 筛选 定义两种 DNMT3A 突变的表型、机制和所赋予的脆弱性 蛋白质的功能域。 Specific Aim 2 将使用能够开启的双重组酶鼠模型 和关闭 DNMT3A 突变，以确定 DNMT3A 突变 CH 的可逆性并评估致癌性 MDS/AML 中 DNMT3A 突变的依赖性。这些研究的意义将带来新的治疗方法 DNMT3A 突变恶性肿瘤的治疗方法。 该提案将在 Ross Levine 博士（发起人）的实验室进行，他是该项目的负责人。 分子癌症医学计划。莱文实验室是人类肿瘤学和发病机制计划的一部分 纪念斯隆凯特琳癌症中心 (MSK) 是一家最先进的癌症研究机构。导师制将 也由 MSK 斯隆表观遗传学中心负责人 Kristian Helin 博士（共同发起人）提供。 这些隶属关系以及格斯特纳·斯隆·凯特林强大的科学和非科学资产 研究生院将提供丰富的协作、技术和科学资源来执行 提议的研究和职业发展。