Microfabricated instrumentation to measure sphingolipid signaling in human acute myeloid leukemia

一种创新的平台来测量单个细胞中鞘磷脂途径的活性 将开发出原发的、人类的、急性髓系白血病(AML)。一个多学科的小组 (化学家、生物工程师、肿瘤学家和计算科学家)有成功的历史 合作将致力于工程化微流控仪器的开发和 使用与医学相关的探头来回答基本问题的支持硬件 关于单个AML细胞的异质性。荧光探针可以同时跟踪 AML细胞中组成神经酰胺-鞘氨酸轴的三条主要途径是 开发的目的是为了详细了解肿瘤细胞中的鞘磷脂信号 已实现。微制造设备内的电泳分离将针对 将单单元测量作为工作流程的一个组成部分。自动化和集成将 极大地提高产量，以生产与常见临床兼容的微设备 工作流程。该提案的一个强大属性是，将执行这些测量 从原始样本中分离出单个细胞，并将避免种群的混淆方面- 散装细胞分析得出的平均数据。此外，通过同时跟踪所有武器 鞘磷脂途径，我们将确定AML细胞动态使用的策略 在药物治疗期间，通过鞘脂信号对它们的促生长途径进行重新编程。 拟议的微细加工装置将在未来提供关于 最佳治疗方案(S)为患者以及产生治疗效果的评估 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.