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

项目摘要 本申请是为了响应特别利益通知（NOSI）而提交的，该通知被确定为 NOT-CA-21-034。急性髓细胞白血病（AML）是最致命的血液癌症之一， 在美国，AML患者每年死亡，表现出臭名昭著的遗传异质性，具有数千种突变 迄今为止在AML患者肿瘤中的描述。AML受益于靶向治疗的兴起， 单个靶向药物的临床影响不大。在AML中，令人印象深刻的初始响应率为60- 在联合使用维奈托克（一种BCL 2抑制剂）和 低甲基化剂。然而，一年的生存率只有30- 40%，这表明需要更多地了解疗效 这种组合。新的离体药物筛选平台已经确定了其他维奈托克组合。 特别是，维奈托克与鲁索利替尼（一种JAK酪氨酸激酶抑制剂）是一种有前途的联合治疗。 基于这一临床前数据，一项新的临床试验已经开始用于复发性或难治性乳腺癌患者。 急性髓细胞白血病 癌症药物组合的决定主要是基于突变异质性，然而， 同基因细胞中非遗传性耐药性的证据越来越多，特别是单细胞分析。 尽管单细胞技术取得了进展，但目前还没有使药物组合的策略。 利用单细胞平台明确解决肿瘤内异质性的决定。借力 CyTOF的优势和解决分析方法的需求，我们开发了一种新的算法 （DRUG-NEM），其分析单个白血病细胞上的单细胞、单药物扰动响应， 为个体患者确定优化的药物组合策略。使用质谱细胞仪分析， 重点关注AML表型、信号传导和代谢，我们将确定对单一药物的离体反应 维奈托克或鲁索利替尼或其组合。使用单药治疗数据，我们将使用DRUG-NEM， 预测对联合用药的反应，并与开始联合用药后试验中获得的患者样本进行比较 疗法如果预测是准确的，它提供了使用单一药物数据的概念证明， 联合治疗，因此是研究未来联合治疗的更有效和实用的方法， 将告知AML患者对维奈托克联合鲁索替尼的敏感性或耐药性。

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.