Developing differentiation therapy for the treatment of patients with acute myeloid leukemia

开发治疗急性髓系白血病患者的分化疗法

• 批准号: 9914232
• 负责人: David B Sykes
• 金额: $ 16.93万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9914232
• 关键词: ATAC-seq; Acute Myelocytic Leukemia; Acute Promyelocytic Leukemia; Adult; Affect; American Society of Hematology; Anabolism; Anthracycline; Basic Science; Biological Assay; Biological Models; Boston; Bypass; CRISPR/Cas technology; Case Study; Cell Culture Techniques; Cell Line; Cells; ChIP-seq; Classification; Clinical; Clinical Trials; Collaborations; Core Facility; Critical Pathways; Cytarabine; Cytotoxic Chemotherapy; DHODH gene; DNA Methylation; DNA Sequence Alteration; Dana-Farber Cancer Institute; Data; Diagnosis; Differentiation Therapy; Dihydroorotate dehydrogenase; Disease; Drug Kinetics; Engineering; Enzymes; Epigenetic Process; Equilibrium; Evaluation; Flow Cytometry; Foundations; Freezing; Gene Expression; Gene Mutation; General Hospitals; Genes; Genetic; Genomics; Goals; Grant; Healthcare; Hematopoietic stem cells; Homeobox; Human; Human Cell Line; Immunotherapy; In Vitro; Inpatients; Institutes; KDM1A gene; Knowledge; Laboratories; Laboratory Research; Learning; Leukemic Cell; Link; Maintenance; Malignant - descriptor; Malignant Neoplasms; Massachusetts; Medical; Mentors; Mentorship; Metabolic; Metabolic Pathway; Microscopy; Modeling; Molecular; Molecular Analysis; Molecular Mechanisms of Action; Muramidase; Mus; Mutation; Myelogenous; Myelopoiesis; Outpatients; Patient Selection; Patients; Pediatric Hospitals; Pharmaceutical Chemistry; Phenotype; Physicians; Positioning Attribute; Post-Translational Protein Processing; Postdoctoral Fellow; Preclinical Drug Development; Preparation; Process; Property; Proteins; Proto-Oncogene Proteins c-akt; Publishing; Pyrimidine; Regenerative Medicine; Reporter; Research; Research Personnel; Resistance; Role; Scientific Advances and Accomplishments; Scientist; Series; Societies; Starvation; Survival Rate; Syndrome; System; Therapeutic; Time; United States National Institutes of Health; Uridine; Work; Xenograft Model; acute myeloid leukemia cell; animal facility; career; chemotherapy; clinical translation; cytotoxicity; design; detection of nutrient; experience; experimental study; gene function; genetic analysis; genome-wide; human model; in vivo; inhibitor/antagonist; leukemia; leukemia initiating cell; leukemia/lymphoma; metabolic profile; mouse model; myeloblast; novel; novel therapeutics; nucleotide metabolism; overexpression; patient subsets; response; screening; senior faculty; small molecule; standard of care; success; teacher; transcription factor; transcriptome sequencing

Project Summary The goal of this project is to develop new, effective, and well-tolerated differentiation therapy for the treatment of patients with acute myeloid leukemia (AML). Despite remarkable advances in understanding the genomic underpinnings of AML, patients still receive the same chemotherapy that they did forty years ago. The study that established the combination of cytarabine and an anthracycline as standard chemotherapy was published in 1973(!). Immunotherapy, and novel target mutations (e.g. IDH, Flt3-ITD, Dot1L) raise the possibility of better therapies for well-defined patient sub- populations. However, differentiation therapy holds the potential of being more globally applicable for leukemias with a wide variety of underlying mutations. The finding that HoxA9 is overexpressed in 70% of AML prompted us to establish a phenotypic screening system to identify small molecules that could overcome the differentiation block established by HoxA9. A high-throughput flow-cytometry differentiation screen, followed by the molecular analysis of compound- resistant cell lines, led to the identification of the enzyme dihydroorotate dehydrogenase (DHODH) as the target of specific inhibitors that could trigger myeloid differentiation. DHODH, and its role in regulating the pool of intracellular uridine, is a novel target in AML. This unexpected in vitro finding in our engineered cell line was confirmed in human cell lines and in ex vivo PDX models of AML. We have since demonstrated that DHODH inhibitors are highly active in vivo in murine syngeneic leukemia models, human xenograft models, and PDX models of AML. This application proposes to determine the molecular mechanism through which the depletion of intracellular uridine results in myeloid differentiation. We have outlined a series of studies that build on preliminary data showing the existence of a clear therapeutic window between normal hematopoietic stem cells and leukemic cells in terms of their ability to survive periods of uridine starvation. While arguably ambitious, the proposed experiments take advantage of completed preliminary studies, allowing us to query the effect of DHODH inhibition in multiple model systems. In particular, the profiling of a 300+ panel of cell lines has yielded lines that are sensitive and lines that are 1000x more resistant to DHODH inhibition. Understanding the basis of this striking phenotype has both basic science and immediate clinical implications in terms of patient selection and disease indication. Understanding the link between pyrimidine biosynthesis and myeloid differentiation will advance our understanding of normal myelopoiesis and of how differentiation is dysregulated in the setting of leukemia. The ready availability of potent inhibitors of DHODH with favorable pharmacokinetic properties raises the exciting possibility of rapid clinical translation. David Sykes is a post-doctoral fellow in Dr. David Scadden’s laboratory at the Massachusetts General Hospital (MGH) Center for Regenerative Medicine. David Scadden is a full professor with a proven track-record of mentoring fellows in the transition to independence. The Center for Regenerative Medicine is a state-of-the-art research laboratory including microscopy and flow-cytometry core facilities as well as its own animal facility. David has established close collaborations with investigators at the Broad Institute, the Dana-Farber Cancer Institute, Boston’s Children’s Hospital, and Bayer Healthcare. These collaborators bring a wealth of co-mentorship to the project as well as scientific expertise in cancer epigenetics, metabolite profiling, medicinal chemistry, and pre-clinical drug development. David’s biosketch speaks to his academic successes. He spent a year as medical chief resident, gaining critical experience as a teacher and a leader. His preliminary research has been recognized with grants from the NIH (R03) as well as the American Society of Hematology, Alex’s Lemonade Stand Foundation, and the Leukemia and Lymphoma Society. He is dedicated to a career as a physician-scientist, and balances outpatient and inpatient clinical responsibilities (20%) with his laboratory research (80%). David is a firm believer that patient encounters help to inform and to guide basic research. To this end, he has identified an entirely new clinical syndrome (the TEMPI syndrome), and written a number of clinical case reports. David looks forward to continued mentorship as he learns to carry out and to interpret experiments in epigenetic and metabolite profiling, and to work with chemists in the rational design of small molecules around targeting critical pathways in differentiation arrest. The next years will build productively on preliminary studies in preparation for transition to an independent investigator position.

项目摘要 该项目的目标是开发新的、有效的、耐受性良好的分化治疗方法。 急性髓系白血病(AML)患者。 尽管在了解急性髓细胞白血病的基因组基础方面取得了显著进展，但患者仍在接受 和他们40年前做的化疗一样。建立阿糖胞苷联合制剂的研究 1973年发表了一种作为标准化疗的蒽环类药物(！)。免疫疗法和新靶点 突变(例如IDH、Flt3-ITD、DOT1L)增加了对明确定义的亚组患者进行更好治疗的可能性 人口。然而，辨证治疗有可能在全球范围内更适用于 具有各种各样潜在突变的白血病。发现HoxA9在70%的AML中高表达 促使我们建立了一种表型筛选系统来识别可以克服 HoxA9建立分化阻滞剂。 高通量流式细胞术分化筛选，然后对化合物- 抗性细胞系，导致二氢罗酸脱氢酶(DHODH)被鉴定为 靶点为可能引发髓系分化的特定抑制物。DHODH及其在调节池子中的作用 胞内尿苷，是AML治疗的新靶点。在我们的工程细胞系中，这一出人意料的体外发现 在人类细胞系和急性髓系白血病的体外PDX模型中得到证实。从那以后我们已经证明了DHODH 抑制物在小鼠同基因白血病模型、人类异种移植模型和PDX中具有高度活性 急性髓系白血病的模型。 这项应用提出了确定细胞内耗竭的分子机制 尿苷可导致髓系分化。我们概述了一系列以初步数据为基础的研究 显示正常造血干细胞和白血病之间存在一个明确的治疗窗口 就细胞在尿苷饥饿期间存活的能力而言。虽然可以说雄心勃勃，但拟议的 实验利用了已完成的初步研究，使我们能够质疑DHODH的效果 多个模型系统中的抑制。特别是，对300多个细胞系面板的剖析已经产生了线 敏感的品系和对DHODH抑制抗性1000倍的品系。理解这一点的基础 突出的表型在患者选择和临床应用方面具有基础科学和直接的临床意义。 疾病指征。 了解嘧啶生物合成和髓系分化之间的联系将促进我们的 了解白血病背景下正常的骨髓生成和分化是如何失调的。这个 具有良好药代动力学特性的DHODH有效抑制剂的现成可提高兴奋性 快速临床翻译的可能性。 大卫·赛克斯是马萨诸塞州总医院大卫·斯卡登博士实验室的博士后研究员 (MGH)再生医学中心。大卫·斯卡登是一名全职教授，在 在向独立过渡的过程中指导同伴。再生医学中心是最先进的 研究实验室包括显微镜和流式细胞仪核心设施以及自己的动物设施。 大卫与博德研究所的研究人员建立了密切的合作关系，达纳-法伯癌症 波士顿儿童医院和拜耳医疗集团。这些合作者带来了丰富的 对该项目的共同指导以及癌症表观遗传学、代谢物图谱、药物 化学和临床前药物开发。 大卫的生平素描说明了他在学术上的成就。他花了一年的时间担任医疗总住院医生，获得了 作为教师和领导者的批判性经验。他的初步研究得到了来自 美国国立卫生研究院(R03)以及美国血液病学会、亚历克斯的柠檬水立场基金会和 白血病和淋巴瘤学会。他致力于作为一名内科科学家的职业，并平衡 门诊和住院的临床责任(20%)与他的实验室研究(80%)。大卫是一家公司 相信患者的接触有助于提供信息和指导基础研究。为此，他确定了一个 全新的临床综合征(Tempi综合征)，并撰写了多篇临床病例报告。 大卫期待着在他学习执行和解释实验的过程中继续得到指导 表观遗传学和代谢物图谱，并与化学家合作，合理设计周围的小分子 针对分化抑制中的关键途径。未来几年将在初步研究的基础上取得丰硕成果 为过渡到独立调查员职位做准备。