MICROFABRICATED INSTRUMENTATION TO MEASURE SPHINGOLIPID SIGNALING IN HUMAN ACUTE MYELOID LEUKEMIA

一个创新的平台来测量单细胞中鞘脂途径的活性， 将发展成原发性人急性髓性白血病（AML）。多学科小组 （化学家，生物工程师，肿瘤学家和计算科学家），具有成功的 合作将致力于开发工程微流体仪器， 支持硬件使用医学相关探头来回答基本问题 关于单个AML细胞的异质性。荧光探针同时追踪 AML细胞中包含神经酰胺-鞘氨醇轴的三个主要途径将是 开发，以便详细了解肿瘤细胞中的鞘脂信号传导， 办妥了一批微加工装置内的电泳分离将针对以下方面进行优化： 单细胞测量作为工作流程的一部分。自动化和集成将 大大提高吞吐量，生产出与常见临床兼容的微型设备 工作流程。该提案的一个强有力的特点是，这些测量将在 对来自原代样品的单细胞，并将避免群体的混淆方面- 通过大量细胞测定得到的平均数据。此外，通过同时跟踪所有武器， 鞘脂途径，我们将确定AML细胞动态使用的策略 在药物治疗期间，通过鞘脂信号转导重新编程其生长促进途径。 所提出的微加工装置将在未来提供关于 为患者提供最佳治疗选择，并对治疗疗效进行评估， 为新兴的精准医疗领域贡献基础数据。

An innovative platform to measure the activity of the sphingolipid pathway in single cells from primary, human, acute myeloid leukemia (AML) will be developed. A multidisciplinary group (chemist, bioengineer, oncologist and computational scientist) with a history of successful collaborations will pursue the development of engineered microfluidic instrumentation and supporting hardware using medically relevant probes to answer fundamental questions regarding heterogeneity in single AML cells. Fluorescent probes to track simultaneously the three major pathways comprising the ceramide-sphingosine axis in AML cells will be developed so that a detailed understanding of sphingolipid signaling in the tumor cells is achieved. Electrophoretic separations within a microfabricated device will be optimized for the single-cell measurements as a component of the work flow. Automation and integration will greatly increase throughput to yield a microdevice which is compatible with common clinical workflows. A powerful attribute of the proposal is that these measurements will be performed on single cells from primary samples and will avoid the confounding aspects of population- averaged data yielded by bulk cell assays. Furthermore, by simultaneously tracking all arms of the sphingolipid pathway, we will identify the strategies that AML cells use to dynamically reprogram their growth-promoting pathways via sphingolipid signaling during drug treatment. The proposed microfabricated devices will in the future provide key information concerning the best treatment option(s) for patients as well yielding an assessment of treatment efficacy to contribute fundamental data to the emerging field of precision medicine.