Targeting mitochondrial dynamics in drug-resistant acute myeloid leukemia

靶向耐药急性髓系白血病的线粒体动力学

• 批准号: 10204995
• 负责人: Christina Glytsou
• 金额: $ 15.3万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204995
• 关键词: Ablation; Acute Myelocytic Leukemia; Adult; Apoptosis; Apoptotic; Architecture; BCL2 gene; Biochemical; Bioinformatics; Blood; CRISPR interference; CRISPR screen; CRISPR/Cas technology; Caspase; Cell Death; Cell Line; Cells; Clinical; Collaborations; Complex Analysis; Crista ampullaris; Data; Development Plans; Drug resistance; Electron Microscopy; Ensure; Equipment; Event; Food; Genes; Genetic; Goals; Growth; Hematopoietic Neoplasms; Hematopoietic stem cells; Homeostasis; Human; Image; In Vitro; Induction of Apoptosis; Institutes; Laboratories; Lead; Malignant Neoplasms; Mediating; Mentors; Microscopy; Mitochondria; Molecular; Morphology; OPA1 gene; Organelles; Pathogenesis; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacology; Phase; Physiological; Process; Prognosis; Proteins; Proteolysis; Proteolytic Processing; Regulation; Research; Research Personnel; Resistance; Resistance development; Resolution; Resources; Role; Sampling; Scientist; Shapes; Stress; Structure; Survival Rate; System; Techniques; Testing; Therapeutic; Training; Xenograft procedure; acute myeloid leukemia cell; base; cancer cell; career development; chaperonin; course development; cytochrome c; drug development; electron tomography; experimental study; genome-wide; in vivo; inhibitor/antagonist; innovation; leukemia; mimetics; mitochondrial dysfunction; mortality; mouse model; novel; post-doctoral training; pre-clinical; prevent; programs; protein complex; research and development; response; skills; small molecule; spatiotemporal; success; targeted treatment; treatment strategy; tumor

PROJECT SUMMARY/ABSTRACT Acute myeloid leukemia (AML) is the second most common leukemia in adults and typically has a dismal prognosis and high mortality, which is exemplified by a 28% five-year overall survival rate. Venetoclax, a selective inhibitor of the anti-apoptotic protein BCL-2, has received FDA approval for the treatment of AML. Despite promising early responses of AML patients to venetoclax, drug resistance ensues after prolonged treatment and highlights the urgency for a deep understanding of the underlying mechanisms. Recently, I discovered that mitochondria in AML cells undergo a morphological change upon venetoclax resistance. Using a genome-wide CRISPRi screen in human AML, I identified genes involved in mitochondrial structure as synthetic lethal vulnerabilities for venetoclax in AML. Mitochondria of venetoclax-resistant AML cells actively modify their architecture and function to prevent apoptosis. Supporting this, OPA1, the master regulator of mitochondrial cristae structure, and CLPB, a mitochondrial chaperonin, were strikingly upregulated in venetoclax-resistant AML cells relative to the sensitive cells. CLPB directly interacts with OPA1 to maintain the physiological mitochondrial morphology. Promisingly, genetic CLPB or OPA1 ablation enhances venetoclax-induced apoptosis of AML cells, by promoting cristae remodeling and mitochondrial stress. This proposal aims to leverage these observations by 1) delineating the mechanistic details by which mitochondrial dynamics and homeostasis lead to acquisition of drug resistance in AML, using super-resolution microscopy, electron tomography, and biochemical techniques, and 2) assessing the therapeutic potential of perturbing mitochondrial structure to augment venetoclax action in preclinical AML mouse models. This research stands to have significant clinical impact, because it can serve as a basis for developing new combinational targeted therapies for patients with AML. In addition, this proposal outlines my career development plan for obtaining the requisite training to transition into a successful independent investigator. This includes 1) guidance from my esteemed mentor Dr. Iannis Aifantis, expert in blood malignancies and mouse models; 2) scientific training by an expert advisory panel, consisting of Drs. Raoul Tibes, Hans-Willem Snoeck, Kivanc Birsoy and Evripidis Gavathiotis, all in top institutes of NYC; 3) hands-on training using state-of-art equipment, including super-resolution microscopy with Dr. Eli Rothenberg; 4) collaboration with experts in microscopy and bioinformatics; and 5) career development courses sponsored by NYU. The laboratory of Dr. Aifantis and NYU Department of Pathology will provide the resources critical to my training and research, ensuring my success. This extensive professional growth program will guide me during the mentored phase excelling as an independent academic scientist. Collectively, the proposed research and career development plans are expected to generate data with significant impact on circumventing targeted-therapy resistance in AML and setting the basis of my future research as an independent researcher.

项目概要/摘要 急性髓系白血病 (AML) 是成人中第二常见的白血病，通常具有令人沮丧的症状 预后差且死亡率高，五年总生存率为 28%。 Venetoclax，一种选择性 抗凋亡蛋白 BCL-2 的抑制剂已获得 FDA 批准用于治疗 AML。尽管 AML 患者对 Venetoclax 有希望的早期反应，但长期治疗后会出现耐药性， 强调了深入了解根本机制的紧迫性。最近，我发现 AML 细胞中的线粒体在维奈托克耐药时会发生形态变化。使用全基因组 人类 AML 中的 CRISPRi 筛选，我鉴定出参与线粒体结构的基因为合成致死基因 Venetoclax 在 AML 中的漏洞。维奈托克耐药 AML 细胞的线粒体主动改变其 防止细胞凋亡的结构和功能。线粒体的主要调节因子 OPA1 支持这一点 嵴结构和 CLPB（一种线粒体伴侣蛋白）在维奈托克耐药 AML 中显着上调 细胞相对于敏感细胞。 CLPB直接与OPA1相互作用，维持线粒体的生理功能 形态学。有望消除基因 CLPB 或 OPA1 增强 Venetoclax 诱导的 AML 细胞凋亡， 通过促进嵴重塑和线粒体应激。 该提案旨在通过 1）描述机制细节来利用这些观察结果 使用超分辨率，线粒体动力学和稳态导致 AML 获得耐药性 显微镜、电子断层扫描和生化技术，以及 2) 评估治疗潜力 扰乱线粒体结构以增强临床前 AML 小鼠模型中的 Venetoclax 作用。这项研究 具有重大的临床影响，因为它可以作为开发新组合的基础 AML 患者的靶向治疗。 此外，该提案概述了我的职业发展计划，以获得过渡所需的培训 成为一名成功的独立调查员。这包括 1) 我尊敬的导师 Iannis 博士的指导 Aifantis，血液恶性肿瘤和小鼠模型专家； 2）专家顾问团的科学培训， 由博士组成。 Raoul Tibes、Hans-Willem Snoeck、Kivinc Birsoy 和 Evripidis Gavathiotis 均来自顶尖研究所 纽约市； 3) 使用最先进的设备进行实践培训，包括 Eli 博士的超分辨率显微镜 罗滕伯格； 4）与显微镜和生物信息学专家的合作； 5）职业发展课程 由纽约大学赞助。 Aifantis博士实验室和纽约大学病理学系将提供资源 对我的培训和研究至关重要，确保我的成功。这项广泛的专业成长计划将指导 我在指导阶段作为一名独立的学术科学家表现出色。总的来说，拟议的 研究和职业发展计划预计将产生对规避风险产生重大影响的数据 AML 的靶向治疗耐药性，并为我作为独立研究者的未来研究奠定了基础。