Dissecting Single-cell Response or resistance to novel combination therapy in AML using mass cytometry

项目摘要(最多30行) 我们开发了一项由研究人员发起的临床试验，以测试鲁索利替尼和万诺克拉克斯的联合应用。 基于广泛的体外小分子的复发/难治性AML患者的(Rux Ven)(NCT03874052) 从数百个原发急性髓系白血病样本中收集的抑制剂筛选数据。初步试验结果显示， 联合用药耐受性好，一些患者在一个周期后得到缓解。我们的协作 与戴维斯博士的提案将利用试验患者的骨髓和外周血样本，这些样本已经被 用每种单一药物和药物组合进行体外治疗，并用单细胞分辨肿块 细胞计数法(CyTOF)。虽然CyTOF可以解决肿瘤内的异质性，在药物筛选的水平上 对于单个单元格，没有处理所产生的复杂数据所需的分析。充分利用优势 并解决了分析方法的需求，普列夫蒂斯博士的团队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。CyTOF分析将检查AML 单个细胞的表型、JAK-STAT信号、BCL2家族蛋白和细胞代谢 患者在研究中接受治疗。我们假设，通过使用药物NEM，我们可以预测对联合治疗的反应 治疗基于体外对单药治疗的反应。我们将把我们预测的反应与 患者的实际反应，并确定与联合耐药相关的细胞特征。至 我们将对收集的患者样本进行细胞因子分析，以确定潜在的耐药机制 整个治疗过程中。我们假设从微环境发出的细胞因子信号可能在 在使用Rux Ven治疗的患者耐药中起关键作用。如果药物-NEM的预测是准确的，它将 为使用单剂药物数据为联合治疗提供概念证明，从而提供更多 为研究未来的联合治疗提供了有效和实用的途径。如果发现特定的细胞因子 对Rux Ven抗性的贡献，可以阐明克服抗性的方法。总的来说，我们的 建议的补充计划利用我们的综合优势来预测药物反应，并进一步 了解AML患者耐药机制。正在提交此申请以请求 为抗药性和敏感性中心(DRSC)提供补充预算，以执行 Kara Davis博士的《行政补充申请》中提出的协作工作 用MASS分析急性髓系白血病患者对新型联合治疗的单细胞反应或耐药 《回应NOT-CA-21-034》特别关注通知(NOSI)：管理 支持与NCI支持的耐药性和敏感度合作的补充 网络(DRSN)。

Project Summary (max 30 lines) We developed an investigator-initiated clinical trial to test the combination or ruxolitinib and venetoclax (Rux+Ven) in relapse/refractory AML patients (NCT03874052) based upon extensive ex vivo small molecule inhibitor screening data collected from hundreds of primary AML samples. Preliminary trial results show the combination is well tolerated with remissions obtained in some patients after a single cycle. Our collaborative proposal with Dr. Davis will make use of marrow and peripheral blood samples from trial patients that have been treated ex vivo with each single agent and the drug combination along with single-cell resolution of mass cytometry (CyTOF). While CyTOF can address intratumoral heterogeneity for drug screening at the level of the single cell, the required analytics for the resulting complex data were not addressed. Leveraging the strengths of CyTOF and addressing the need for analysis approaches, Dr. Plevritis’ group 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 individual patients. The CyTOF analysis will examine AML phenotype, JAK-STAT signaling, BCL2 family proteins and cellular metabolism on individual cells from the patients treated on study. We hypothesize that by using DRUG-NEM we can predict response to combination treatment based on in vitro response to single agent treatment. We will compare our predicted response to the patient’s actual response and determine cellular features associated with resistance to the combination. To identify potential mechanisms of resistance we will perform cytokine assays on patient samples collected throughout treatment. We hypothesize that cytokine signals emanating from the microenvironment could play a key role in drug resistance in patients treated with Rux+Ven. If the DRUG-NEM predictions are accurate, it will provide proof of concept for using single-agent drug data to inform combination therapy, thus, providing a more efficient and practical approach to study future combination treatments. If specific cytokines are found to contribute to Rux+Ven resistance, approaches to overcome the resistance can be elucidated. Overall, our proposed supplement project makes use of our combined strengths to predict drug response and further understand mechanisms of drug resistance in AML patients. This application is being submitted to request a supplemental budget for the Drug Resistance and Sensitivity Center (DRSC) to perform the collaborative work as proposed in Dr. Kara Davis’ administrative supplement application titled "Dissecting Single-Cell Response or Resistance to Novel Combination Therapy in AML Using Mass Cytometry ” in response to NOT-CA-21-034 “Notice of Special Interest (NOSI): Administrative Supplements to Support Collaborations with the NCI-supported Drug Resistance and Sensitivity Network (DRSN).