Genetics and Signaling of Drug Resistance and Sensitivity in AML Cell Lines, Xenografts, and Primary Patient Samples

AML 细胞系、异种移植物和主要患者样本中耐药性和敏感性的遗传学和信号转导

• 批准号: 10038083
• 负责人: BRIAN J DRUKER
• 金额: $ 34.85万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10038083
• 关键词: Acute Myelocytic Leukemia; Automobile Driving; BCL2 gene; Biochemical; Cell Line; Cells; Cellular biology; Chronic Myeloid Leukemia; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Computer Models; Custom; Data; Disease; Drug Combinations; Drug Screening; Drug Synergism; Drug Targeting; Drug resistance; Essential Genes; Evaluation; FLT3 gene; FLT3 inhibitor; Feedback; Gene Mutation; Gene Targeting; Genes; Genetic; Genetic Models; Genomics; Goals; Imatinib; JAK1 gene; Lesion; Libraries; MEKs; Malignant Neoplasms; Methods; Modeling; Molecular; Mus; Mutation; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Resistance; Resistance development; Role; Route; Sampling; Signal Pathway; Signal Transduction; Survival Rate; Techniques; Testing; Validation; Work; Xenograft Model; Xenograft procedure; acute myeloid leukemia cell; base; bcr-abl Fusion Proteins; clinically relevant; cohort; drug candidate; drug sensitivity; experimental study; functional genomics; genome-wide; genomic data; genomic locus; improved; improved outcome; inhibitor/antagonist; insight; kinase inhibitor; knowledge base; leukemia; multiple omics; neoplastic cell; next generation sequencing; novel drug combination; novel therapeutics; personalized medicine; pre-clinical; prognostic; programs; resistance mechanism; resistance mutation; response; screening; success; targeted agent; therapeutic target; tool; treatment strategy; tumor

Acute myeloid leukemia (AML), a highly lethal subtype of leukemia, has a 5-year survival rate of <20%. With the advent of next-generation sequencing, a number of specific genetic lesions that drive AML and provide prognostic information have been identified. Previous work by our group (co-PD/PI, Druker) provided proof that imatinib, a targeted ABL kinase inhibitor that blocks activity of the BCR-ABL fusion protein in chronic myeloid leukemia (CML), dramatically improves patient survival. Similar approaches have been applied to AML, a more genetically complex leukemia, and some drugs have improved outcomes, but none have been as successful as ABL inhibitors for CML due to incomplete responses and the rapid development of resistance. Project 1 of this DRSC Program, Drug Combinations to Circumvent Resistance (D2CR), will focus on understanding intrinsic mechanisms of enhanced drug sensitivity or resistance, with the goal of devising novel therapeutic strategies. For this Project, our long-term goals are to nominate drugs that enhance upfront drug sensitivity and/or circumvent resistance for use in combination strategies that will be tested in clinical trials. Our immediate goals are to identify essential target genes and pathways contributing to sensitivity or resistance to specific drugs and to validate their roles using cell lines, patient samples, and xenograft-derived cells. These goals are based on our central hypothesis that the heterogeneous genetic landscape of AML, in tandem with complex signaling feedback loops, contributes to intrinsic mechanisms of drug sensitivity and resistance. Project 1 will provide critical preclinical data to advance drug candidates for use in combinations tested in primary patient samples and xenograft models in Project 3. To accomplish these goals, 3 Aims are proposed: 1) Identify genetic mechanisms of drug sensitivity/resistance in AML cell lines through essential-gene and drug-resistance screens – We will perform genome-wide CRISPR/Cas essential gene and re-sensitization screens to identify gene targets and pathways contributing to enhanced drug activity or resistance. Data generated by the combination of these 2 screens will provide key insights into cell-intrinsic mechanisms of sensitivity or resistance to 5 select drugs (crenolanib, quizartinib, ruxolitinib, trametinib, and venetoclax) in AML cells with diverse genetic backgrounds. 2) Computationally validate, refine, and inform candidate pathways and genes contributing to intrinsic mechanisms of drug sensitivity or resistance – This iterative modeling step will leverage the intrinsic genetic factors identified by our in-house Cancer Targetome Knowledgebase and will prioritize targets for further validation in Aim 3. 3) Validate new gene targets in hypothesis-driven, focused CRISPR/Cas experiments – We will develop a targeted CRISPR/Cas sgRNA library to perturb genes hypothesized to contribute to drug sensitivity or resistance in cell line models, xenografts, and primary patient samples. We expect to identify gene targets for further interrogation in drug combination testing in Project 3.

急性髓系白血病（AML）是一种高度致命的白血病亚型，5年生存率<20%。和 下一代测序的出现，一些驱动 AML 的特定基因损伤并提供 预后信息已确定。我们小组（联合 PD/PI、Druker）之前的工作证明了这一点 伊马替尼，一种靶向 ABL 激酶抑制剂，可阻断慢性骨髓细胞中 BCR-ABL 融合蛋白的活性 白血病（CML），显着提高患者的生存率。类似的方法已应用于 AML，这是一种更 遗传复杂的白血病，一些药物可以改善结果，但没有一个如此成功 作为 CML 的 ABL 抑制剂，由于反应不完全和耐药性的快速发展。项目1的 这个 DRSC 计划，即规避耐药性的药物组合 (D2CR)，将重点关注了解 增强药物敏感性或耐药性的内在机制，目的是设计新的治疗方法 策略。对于这个项目，我们的长期目标是提名能够增强前期药物的药物 敏感性和/或规避抗性用于将在临床中进行测试的组合策略 试验。我们的近期目标是确定重要的靶基因和途径，以促进 对特定药物的敏感性或耐药性，并使用细胞系、患者样本验证其作用， 和异种移植衍生的细胞。这些目标基于我们的中心假设，即异质性 AML 的遗传景观与复杂的信号反馈环相结合，有助于内在的 药物敏感性和耐药性的机制。项目1将提供关键的临床前数据以推进 在项目 3 中，在主要患者样本和异种移植模型中测试组合使用的候选药物。 为了实现这些目标，提出了 3 个目标：1）确定药物敏感性/耐药性的遗传机制 通过必需基因和耐药性筛选在 AML 细胞系中进行筛选 – 我们将进行全基因组筛选 CRISPR/Cas 必需基因和再敏化筛选，以确定基因靶点和通路 增强药物活性或耐药性。这两个屏幕组合生成的数据将提供关键 深入了解对 5 种精选药物（crenolanib、quizartinib、 鲁索替尼、曲美替尼和维奈托克）在具有不同遗传背景的 AML 细胞中的作用。 2）计算上 验证、完善并告知有助于药物内在机制的候选途径和基因 敏感性或抵抗力——这个迭代建模步骤将利用我们确定的内在遗传因素 内部癌症目标组知识库，并将优先考虑目标，以便在目标 3 中进一步验证。3) 在假设驱动的、集中的 CRISPR/Cas 实验中验证新的基因靶标 – 我们将开发 靶向 CRISPR/Cas sgRNA 文库扰乱推测有助于药物敏感性的基因或 细胞系模型、异种移植物和主要患者样本中的耐药性。我们期望确定基因靶标 项目3中药物组合测试的进一步询问。