Microfluidic Devices for Studying Cancer Signal Transduction

描述(申请人提供)：各种癌症的出现和发展受到细胞外信号的严重影响，如生长因子和炎性细胞因子。由这些信号激活的细胞内信号通路通常会改变行为和新的动态特性，导致肿瘤行为。此外，在一个特定的细胞中，多个信号通路可能被激活，导致复杂的相互作用，称为‘串扰’，并使其难以确定关键的致癌途径，或提出有针对性的治疗。因此，需要对癌症信号有一个全面的、系统范围的看法。 实现对癌细胞动态信号行为的真正系统分析的一个重要障碍是缺乏系统、有效和可重复地测量即时信号输出的工具。在这里，我们建议开发和增强能够克服这一障碍的新型微流控器件。这些设备包含数十个可以装满电池的微型室。使用传统的免疫细胞化学方法，每个腔室中的细胞都可以被独立地刺激和监测，以确定不同的信号事件。除了可以在单个设备中进行大量并行测量外，微流体还提供了对实验条件的精确控制，增强了重复性，使定量更加准确。我们现有的原型已经过所有主要必需功能的测试，并为我们的技术方法提供了原则证明。 我们建议进一步开发现有的原型，以便能够进行非常高吞吐量的实验，对不同癌细胞中的细胞信号特征进行分类。这项应用的主要重点是增加设备和辅助设备的尺寸和容量，并通过对癌细胞系和恶性黑色素瘤活检细胞的信号进行中试筛选来展示其功能。这些试点筛选可能揭示新的基于信号转导的癌症诊断标记物，并建议最佳的化疗靶点。我们预计，该装置将成为研究信号转导的强大研究工具。此外，该设备的使用可能会转化为临床实验室，特别是它直接在患者细胞上进行多项实验的能力。我们预计临床医生最终将能够在个性化治疗中使用该设备，例如，通过使用小活检来筛选标记药物敏感性的各种信号特征，或通过直接测试化疗药物的疗效。

DESCRIPTION (provided by applicant): The emergence and progression of various cancers is heavily influenced by extracellular cues such as growth factors and inflammatory cytokines. The intracellular signaling pathways activated by these cues often have altered behavior and new dynamic properties, leading to neoplastic behavior. Furthermore, in a given cell, multiple signaling pathways may be activated leading to complex interactions known as 'cross-talk' and making it difficult to identify key carcinogenic pathways, or to propose targeted treatment. Therefore, a comprehensive, systems-wide view of cancer signaling is required. An important barrier to achieving a truly systemic analysis of dynamic signaling behavior in cancer cells is the lack of a tool to systematically, efficiently, and reproducibly measure the immediate signaling outputs. Here we propose to develop and enhance novel microfluidic devices that can overcome this barrier. The devices contain dozens of miniature chambers which can be filled with cells. Cells in each chamber can be independently stimulated and monitored for a distinct signaling event using conventional immunocytochemistry methods. In addition to the large number of parallel measurements that can be made in a single device, microfluidics offers precise control over experimental conditions, enhancing reproducibility and making quantification more accurate. Our existing prototypes have been tested for all major required functions and provide proof-of-principle of our technological approach. We propose to further develop the existing prototypes to enable very high-throughput experiments cataloging cell signaling signatures in diverse cancer cells. The main focus of this application is to increase the size and capacity of the device and supporting equipment, and demonstrate its functionality through a pilot screen of signaling in cancer cell lines and cells from malignant melanoma biopsies. These pilot screens may reveal new signal transduction-based markers for cancer diagnosis and suggest optimal chemotherapeutic targets. We anticipate that the device will become a powerful research tool to study signal transduction. Furthermore, the use of the device could potentially translate to clinical laboratories, specifically in its ability to perform multiple experiments directly on a patient's cells. We envision clinicians will eventually be able to use the device in personalized therapy, for example, by using a small biopsy to screen for various signaling features that mark drug susceptibility or by directly testing the efficacy of chemotherapeutic agents.