Mechanisms and In Vivo Efficacy of Synergistic Acid Ceramidase and Bcl-2 Inhibition in Acute Myeloid Leukemia

酸性神经酰胺酶和 Bcl-2 协同抑制治疗急性髓系白血病的机制和体内疗效

• 批准号: 10743571
• 负责人: Johnson Ung
• 金额: $ 3.83万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2029-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10743571
• 关键词: Academia; Acute Myelocytic Leukemia; Affect; Apoptosis; Apoptotic; BCL2 gene; BCL2L1 gene; Bioinformatics; Biological Assay; Biological Markers; Bone marrow failure; Breast Cancer Model; Breast Cancer Treatment; CRISPR screen; CRISPR/Cas technology; Cancer Research Project; Cell Death; Cell Line; Cell Membrane Permeability; Ceramides; Clinical; Combined Modality Therapy; Data; Data Analyses; Drug Combinations; Electron Microscopy; Engraftment; Enzymes; Equilibrium; FDA approved; Family; Genetic; Goals; Grant; Human; Hydrolase; Impairment; In Vitro; Isopropyl Thiogalactoside; Knock-out; Learning; Link; Lipids; MCL1 gene; Malignant Neoplasms; Mediating; Mentors; Metabolism; Methods; Mitochondria; Modality; Mus; Myelogenous; Myeloproliferative disease; N-caproylsphingosine; Newly Diagnosed; Outcome; Outer Mitochondrial Membrane; Patients; Pharmaceutical Preparations; Phase; Postdoctoral Fellow; Productivity; Protein Family; Proteins; Relapse; Research; Research Personnel; Research Project Grants; Resistance; Respiration; Role; Sampling; Series; Signal Transduction; Solid Neoplasm; Sphingolipids; Testing; Therapeutic; Training; Treatment Efficacy; Up-Regulation; Work; acute myeloid leukemia cell; analog; antagonist; anticancer research; bcl-xlong protein; biological adaptation to stress; biomarker identification; cancer therapy; career; chemotherapy; combinatorial; design; drug sensitivity; galactosylgalactosylglucosylceramidase; genomic data; improved; in vivo; inhibitor; knock-down; leukemia; leukemia treatment; malignant breast neoplasm; member; mimetics; molecular targeted therapies; nanoliposome; novel; overexpression; pharmacologic; pre-doctoral; precision medicine; preclinical efficacy; prognostic value; programs; protein expression; respiratory; response; skills; sphingosine 1-phosphate; synergism; therapeutic target; transcriptome sequencing; transcriptomics; treatment strategy; triple-negative invasive breast carcinoma; vector; whole genome

PROJECT SUMMARY/ABSTRACT Acute myeloid leukemia (AML) is an aggressive, heterogeneous myeloid malignancy that results in bone marrow failure. The Bcl-2 antagonist/BH3-mimetic, venetoclax (VEN), is used in combination with chemotherapy or hypomethylating agents to treat newly diagnosed AML patients unable to tolerate induction chemotherapy. However, relapse rates remain high due to de novo and acquired resistance attributed to mitochondrial reprogramming and upregulation of compensatory survival proteins like Mcl-1 and Bcl-xL. Strategies that antagonize mitochondrial respiration, induce the integrated stress response (ISR), and/or reduce Mcl-1/Bcl-xL protein expression are known to improve VEN sensitivity. Importantly, sphingolipid (SL) metabolism, mitochondrial respiration, the ISR, and Mcl-1 protein levels are closely linked, making SL metabolism an attractive therapeutic target in combination with VEN. The balance of pro-apoptotic ceramides and pro-survival sphingosine-1-phosphate form the bioactive core of sphingolipid signaling. Acid ceramidase (AC) is a lipid hydrolase that catabolizes pro-apoptotic ceramides. We previously demonstrated that AML blasts rely on AC for survival. Because we and others showed that AC inhibition severely impairs mitochondrial respiration, we sought to determine whether combining AC inhibitors with VEN could be effective for treating AML. Our preliminary data show that AC inhibition improves VEN sensitivity in human AML cell lines and patient samples. However, the mechanisms underlying the synergy from this novel combination are unknown, and the in vivo efficacy of combinatorial AC and Bcl-2 inhibition has not been explored. We hypothesize that synergy between AC and Bcl- 2 inhibition results from SL-mediated impairment of mitochondrial form, respiration, and ISR overactivation. Aim 1 (F99 phase) is designed to complete my dissertation work by characterizing the mechanism, biomarkers, and in vivo efficacy of combined AC and Bcl-2 inhibition in AML. To accomplish these goals, we will analyze mitochondrial form and function, interrogate the role of the ISR for synergy, and utilize computational approaches to define biomarkers of drug responses. My overarching career goal is to develop into a highly productive, independent cancer researcher focused on developing novel SL-based therapeutics and precision medicine. To best prepare myself to start an independent cancer research program, I will utilize the K00 period to i) learn new skills in dynamic BH3 profiling (a precision medicine assay), CRISPR-Cas9 screening, and genomic data analysis; and ii) expand my cancer research portfolio into solid tumors by studying triple-negative breast cancer. Thus, Aim 2 (K00 phase) will evaluate the relationship between SLs and mitochondrial apoptotic priming using dynamic BH3 profiling and will utilize CRISPR screening to identify novel SL-based combination therapies to treat triple-negative breast cancer. The training plans in the F99/K00 phases will help me successfully achieve my career goals and thrive as an independent cancer researcher.

项目总结/摘要 急性髓系白血病（AML）是一种侵袭性、异质性骨髓恶性肿瘤， 失败Bcl-2拮抗剂/BH3模拟物维奈托克（VEN）与化疗或化疗药物联合使用。 低甲基化剂治疗不能耐受诱导化疗的新诊断AML患者。 然而，由于线粒体损伤引起的新生和获得性耐药，复发率仍然很高。 补偿性存活蛋白如Mcl-1和Bcl-xL的重编程和上调。战略 拮抗线粒体呼吸，诱导综合应激反应（ISR），和/或降低Mcl-1/Bcl-xL 已知蛋白质表达提高VEN敏感性。重要的是，鞘脂（SL）代谢， 线粒体呼吸、ISR和Mcl-1蛋白水平密切相关，使SL代谢和 有吸引力的治疗靶点与VEN组合。促凋亡神经酰胺和促存活神经酰胺的平衡 鞘氨醇-1-磷酸形成鞘脂信号传导的生物活性核心。酸性神经酰胺酶（AC）是一种脂质 分解代谢促凋亡神经酰胺的水解酶。我们以前证明AML原始细胞依赖于AC， 生存因为我们和其他人表明AC抑制严重损害线粒体呼吸，我们寻求 以确定AC抑制剂与VEN组合是否可有效治疗AML。我们的初步数据 显示AC抑制改善了人AML细胞系和患者样品中VEN敏感性。但 这种新组合的协同作用机制尚不清楚， AC和Bcl-2的组合抑制尚未被探索。我们假设AC和Bcl-2之间的协同作用- 2抑制结果SL介导的线粒体形式，呼吸和ISR过度激活的损害。 目的1（F99期）旨在完成我的论文工作，通过表征机制，生物标志物， 以及AC和Bcl-2联合抑制在AML中的体内功效。为了实现这些目标，我们将分析 线粒体的形式和功能，询问ISR的协同作用，并利用计算方法 来定义药物反应的生物标志物。我的总体职业目标是发展成为一个高效， 独立癌症研究人员专注于开发基于SL的新疗法和精准医学。到 最好准备自己开始一个独立的癌症研究计划，我将利用K00期间i）学习新的 在动态BH3分析（一种精确的医学分析）、CRISPR-Cas9筛选和基因组数据方面的技能 分析; ii）通过研究三阴性乳腺癌，将我的癌症研究组合扩展到实体瘤。 因此，Aim 2（K00期）将使用以下方法评估SL与线粒体凋亡引发之间的关系： 动态BH3谱分析，并将利用CRISPR筛选来确定新的基于SL的联合疗法， 治疗三阴性乳腺癌F99/K00阶段的培训计划将帮助我成功实现 我的职业目标，并作为一个独立的癌症研究人员茁壮成长。