Dissecting Single-cell Response or Resistance to Novel Combination Therapy in AML using Mass Cytometry

项目总结 本申请是对特别利益通知(NOSI)的回应，该通知被确认为 不是-CA-21-034。急性髓系白血病(AML)是最致命的血癌之一，超过10,000人 美国每年死亡的患者显示出臭名昭著的基因异质性，有数千个突变 迄今为止在急性髓系白血病患者肿瘤中的描述。AML受益于靶向治疗的兴起，尽管 个别靶向药物的临床影响并不大。在急性髓细胞白血病中，令人印象深刻的初始应答率为60- 80%的老年患者联合使用bcl2抑制剂文奈德和 低甲基化剂。然而，一年的存活率只有30%-40%，建议更多地了解其疗效 这种组合。新的体外药物筛选平台已经确定了更多的万乃馨组合。 特别是，文奈德联合JAK酪氨酸激酶抑制剂鲁索利替尼是一种很有前途的联合治疗方法。 基于这些临床前数据，一项针对复发或难治性患者的新的临床试验已经启动。 AML。 抗癌药物组合的决定主要是基于突变的异质性，然而 越来越多的证据表明，同基因细胞之间存在非遗传耐药性，特别是在单细胞分析方面。 尽管单细胞技术取得了进步，但目前还没有制造药物组合的策略 利用单细胞平台明确解决肿瘤内异质性的决定。利用 针对细胞周期分析的优点和分析方法的需要，我们开发了一种新的算法 (Drug-NEM)分析单个白血病细胞上的单细胞、单药物扰动反应，以 确定针对个别患者的最佳药物组合策略。使用质量细胞术分析 关注AML的表型、信号转导和代谢，我们将确定单一药物的体外反应 静脉滴注或鲁索利替尼或其组合。利用单药治疗数据，我们将使用药物-NEM 预测对联合的反应，并与开始联合后在试验中获得的患者样本进行比较 心理治疗。如果预测是准确的，它为使用单剂药物数据提供概念证明 因此，联合治疗是研究未来联合治疗和治疗的更有效和更实用的方法 将告知急性髓系白血病患者对万乃馨与鲁索利替尼联合使用的敏感性或耐药性。

PROJECT SUMMARY This application is being submitted in response to the Notice of Special Interest (NOSI) identified as NOT-CA-21-034. Acute myeloid leukemia (AML) is among the deadliest blood cancers with over 10,000 patients dying annually in the U.S. AML displays notorious genetic heterogeneity with thousands of mutations described across AML patient tumors to date. AML has benefited from the rise of targeted therapies although clinical impact of individual targeted agents has been modest. In AML, impressive initial response rates of 60- 80% have been observed in elderly patients using the combination of venetoclax, a BCL2 inhibitor, and hypomethylating agents. Yet, survival at one year is only 30-40%, suggesting more to learn about the efficacy of this combination. Novel ex vivo drug screening platforms have identified additional venetoclax combinations. In particular, venetoclax with ruxolitinib, a JAK tyrosine kinase inhibitor, is a promising combination therapy. Based on this preclinical data, a novel clinical trial has been initiated for patients with relapsed or refractory AML. Cancer drug combination decisions are primarily made on the basis of mutational heterogeneity, yet evidence of non-genetic drug resistance among isogenic cells is mounting, particularly with single cell analysis. Despite advances in single cell technologies, there are currently no strategies for making drug combination decisions that utilize single cell platforms to explicitly address intratumoral heterogeneity. Leveraging the strengths of CyTOF and addressing the need for analysis approaches, we developed a novel algorithm (DRUG-NEM) that analyzes single-cell, single-drug perturbation responses on individual leukemia cells to identify optimized drug combination strategies for the individual patient. Using mass cytometry analysis with focus on AML phenotype, signaling and metabolism, we will determine ex vivo response to single agent venetoclax or ruxolitinib or the combination. Using the single agent treatment data, we will use DRUG-NEM to predict response to the combination and compare to patient samples obtained on trial after staring combination therapy. If predictions are accurate, it provides proof of concept for using single-agent drug data to inform combination therapy, thus a more efficient and practical approach to study future combination treatments and will inform sensitivity or resistance to venetoclax in combination with ruxolitinib in patients with AML.