Targeting PML for therapy in leukemia-initiating cells

针对 PML 治疗白血病起始细胞

• 批准号: 8525793
• 负责人: Keisuke Ito
• 金额: $ 24.9万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-20 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8525793
• 关键词: Ablation; Acute; Acute Promyelocytic Leukemia; Address; Arsenic; Arsenic Trioxide; Award; Blast Cell; Bone Marrow; Cell Cycle Regulation; Cell Maintenance; Cell Proliferation; Cells; Chronic Myeloid Leukemia; Clinic; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Dasatinib; Data; Defect; Development; Disease; Disease Reservoirs; Doctor of Philosophy; Effectiveness; Environment; Evaluation; Failure; Genes; Genetic; Goals; Hematologic Neoplasms; Hematologist; Hematopoietic stem cells; Imatinib; Imatinib mesylate; In Vitro; Israel; K-Series Research Career Programs; Knockout Mice; Knowledge; Lead; Maintenance; Malignant - descriptor; Malignant Neoplasms; Mediating; Medical center; Mentors; Modeling; Molecular; Mus; Nature; Osteocalcin; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Play; Population; Pre-Clinical Model; Program Development; Property; Proteins; Recurrent disease; Research; Research Personnel; Research Project Grants; Residual Tumors; Resistance; Resources; Role; Safety; Scientist; Sirolimus; Solid; Solid Neoplasm; Stem cells; Therapeutic; Training; Training Programs; Transgenic Mice; Translating; Tumor Suppressor Proteins; Tyrosine Kinase Inhibitor; University Hospitals; Work; anticancer research; base; cancer stem cell; career; cell growth; chemotherapy; clinically relevant; efficacy testing; experience; human FRAP1 protein; improved; in vivo; leukemia; leukemic stem cell; mTOR inhibition; mouse model; multicatalytic endopeptidase complex; novel therapeutic intervention; pre-clinical; preclinical study; prevent; programs; self-renewal; skills; stem cell biology; success; therapeutic target; tool; tumor

6- PROJECT SUMMARY This proposal describes a five-year training program for the development of an academic career in Cancer Stem Cell Biology. I have four-years of experience as a cancer biologist, two-years as a stem cell biologist, and five-years as a hematologist in a university hospital. I am now planning to expand my knowledge, skills and research field to cancer-initiating cells by combining stem cell and cancer research. My goal over the next five years is to become an independent scientist in the field of cancer stem cell biology, especially in leukemia-initiating cells. I am planning to reach my goal by a two-phase plan. The first phase (first 2-3 years) will be characterized by intense bench work to construct the basis of my research projects and to acquire experience in fields that are new to me, such as the characterization of hematological malignancies in clinically relevant mouse models and contribution of clinical trials in the U.S.A. Further, I will develop/establish niche-specific Pml-deleted mouse models for my own research during independent phase. In the first part of the program, I will be mentored by Pier Paolo Pandolfi, PhD, a leading cancer biologist who has great experience in modeling leukemia and solid tumors in vivo and who has trained numerous independent investigators. The program will be enriched by the collaboration of Drs. D.G. Tenen, D.E. Avigan, T. Suda and J. Teruya-Feldstein. The first part of the award will be completed at Beth Israel Deaconess Medical Center, a scientific environment of excellence with the solid clinical resources necessary to promote the success of this Career Development Award. The second phase will be entirely devoted to establishing my career as an independent scientist. One of the main topics in this phase would be detailed functional analysis of Pml in the microenvironment in niche-specific conditional knockout mice. Five years of support are necessary to produce sufficient data, especially in consideration of the mouse models that I am planning to establish, to allow me to pursue an independent career track. Research will focus on the study of the mechanisms regulating quiescence in leukemia-initiating cells (LICs). Specifically, I am aiming at the development of LIC-specific therapy in paradigmatic hematopoietic stem cell (HSC) disease: chronic myeloid leukemia (CML). Maintenance of leukemia has been demonstrated to be dependent upon a small sub-population of cells within the bulk leukemic population that have self-renewal properties and are termed "leukemia-initiating cells" (LICs). LICs share mechanistic properties with regular stem cells including a more quiescent nature, which is thought to mediate their resistance to standard chemotherapy-based treatment. Failure to effectively target LICs can result in disease relapse. Chronic Myelogenous Leukemia (CML) is an extensively studied stem cell disorder in which the LIC pool is not always eradicated by current targeted therapy, leading to disease relapse upon drug discontinuation. I have defined the essential role of PML in the maintenance of CML-initiating-cell, and present a new therapeutic approach for targeting quiescent LICs by pharmacological inhibition of PML. I have demonstrated that expression of the Promyelocytic Leukemia (PML) tumor suppressor is surprisingly high in both regular hematopoietic stem cells (HSCs) and in CML blasts and that loss of PML expression predicts a more favorable outcome in CML. I subsequently demonstrated that PML plays a key role in maintaining the quiescence and self-renewal properties of HSCs/LICs although the exact molecular mechanisms involved are poorly understood. PML is also known to be pharmacologically inhibited by treatment of cells with arsenic, which specifically decreases the stability of the protein. Taking advantage of this, I have used arsenic-mediated ablation of Pml in a mouse model of CML to successfully target LICs. This work has the potential to have a significant impact on treatment and eradication of CML. Importantly, my findings support the notion PML-ablation by arsenic might be an effective tool to render CML-initiating cells more sensitive to anti-tumor therapy. However, the effectiveness of PML targeting both in clinically relevant mouse model and in the clinic needs to be assessed. Therefore, in order to understand the key pathways downstream PML required for LIC maintenance and to translate PML-ablative LIC targeting to the clinic, I propose the following Specific Aims: (1) to further define the mechanisms of PML-dependent cell cycle regulation in LICs; (2) to analyze the effect of PML loss in the interaction between LICs and their niche; (3) to test the efficacy of arsenic-mediated therapy in preclinical models and finally to support a clinical trial of combination arsenic + Dasatinib treatment for CML through in vitro evaluation. The Training Program outlined in this proposal will launch my independent research career. Importantly, accomplishing these aims could greatly improve treatment of patients with CML, possibly allowing discontinuation of therapy after LIC eradication. Further, understanding PML function could provide other therapeutic targets for LIC and cancer stem cell ablation.

6-项目摘要 该提案描述了一个为期五年的培训计划，用于发展癌症干细胞的学术生涯 细胞生物学我有四年的癌症生物学家经验，两年的干细胞生物学家经验， 在一家大学医院做了五年的血液学家我现在计划扩大我的知识，技能和 通过干细胞和癌症研究相结合，将研究领域扩展到癌症起始细胞。 我在未来五年的目标是成为癌症干细胞生物学领域的独立科学家， 尤其是在白血病起始细胞中。我计划通过两阶段计划来实现我的目标。第一阶段（First 2-3年）的特点将是紧张的板凳工作，以构建我的研究项目的基础， 在对我来说是新的领域获得经验，如血液恶性肿瘤的表征， 临床相关的小鼠模型和美国临床试验的贡献。此外，我将开发/建立 小生境特异性Pml缺失小鼠模型，用于我自己独立阶段的研究。第一部分 计划，我将由Pier Paolo Pandolfi博士指导，Pier Paolo Pandolfi博士是一位领先的癌症生物学家，他在 在体内对白血病和实体瘤进行建模，并培训了许多独立的研究人员。的 该计划将通过D.G.博士的合作来丰富。Tenen，D.E.阿维根，T.苏达和J. Teruya-Feldstein。 该奖项的第一部分将在贝丝以色列女执事医疗中心完成，这是一个科学的环境， 卓越与必要的坚实的临床资源，以促进这个职业发展奖的成功。 第二阶段将完全致力于建立我的职业生涯作为一个独立的科学家。的一个主要 本阶段的主题是特定于利基的微环境中Pml的详细功能分析 条件性基因敲除小鼠五年的支持是必要的，以产生足够的数据，特别是在 考虑到我计划建立的小鼠模型，允许我追求独立的职业生涯 轨道 研究将集中于白血病起始细胞（LIC）中调节静止的机制的研究。 具体来说，我的目标是在典型的造血干细胞中开发LIC特异性治疗。 (HSC)慢性粒细胞白血病（CML） 白血病的维持已被证明依赖于细胞内的一小群细胞。 具有自我更新特性的大量白血病细胞群，被称为“白血病起始细胞”（LIC）。 LIC与常规干细胞具有共同的机械特性，包括更静止的性质，这被认为是 介导其对标准化疗治疗的抗性。如果不能有效地针对低收入国家， 在疾病复发。慢性粒细胞白血病（CML）是一种被广泛研究的干细胞疾病，其中 目前的靶向治疗并不总能根除LIC，导致药物治疗后疾病复发 中止 我已经确定了PML在CML起始细胞维持中的重要作用，并提出了一种新的治疗方法， 通过药物抑制PML靶向静止LIC的方法。我已经证明了 早幼粒细胞白血病（PML）肿瘤抑制因子的表达在两种常规白血病中都令人惊讶地高。 造血干细胞（HSC）和CML母细胞中PML表达缺失预示着更有利的 CML的结果。随后，我证明了PML在维持静止中起着关键作用， HSC/LIC的自我更新特性，尽管所涉及的确切分子机制知之甚少。 还已知用砷处理细胞可抑制PML， 降低了蛋白质的稳定性。利用这一点，我已经在一项研究中使用了砷介导的Pml消融。 CML小鼠模型成功靶向LIC。 这项工作有可能对CML的治疗和根除产生重大影响。重要的是我 研究结果支持砷的PML消融可能是一种有效的工具，使CML起始细胞更多地 对抗肿瘤治疗敏感。然而，PML靶向临床相关小鼠的有效性 模型和临床需要进行评估。因此，为了了解下游的关键途径 LIC维持所需的PML并将PML消融LIC靶向转化为诊所，我建议 以下具体目的：（1）进一步确定LIC中PML依赖性细胞周期调控的机制;（2） 分析PML缺失在LIC与生态位相互作用中的作用;（3）检验 在临床前模型中进行砷介导的治疗，并最终支持砷+ 通过体外评价达沙替尼治疗CML。 本建议书中概述的培训计划将启动我的独立研究生涯。 重要的是，实现这些目标可以大大改善CML患者的治疗， LIC根除后停止治疗。此外，了解PML功能可以提供其他 LIC和癌症干细胞消融的治疗靶点。