A Droplet-Based System for Capture, Manipulation, and Biochemical Profiling of Ra

描述(由申请人提供)：我们认为，开发一种可靠和强大的技术来有效地检测、表征和直接检测微小残留病(MRD)，包括来自血液的循环肿瘤细胞(CTCs)和来自骨髓的播散性肿瘤细胞(DTC)，将对阐明转移过程的生物学和开发癌症管理和治疗的新方法做出重大贡献。这些细胞是癌症传播和致死的重要因素。癌症是一种令人沮丧的疾病--在五年内，近30%的癌症患者死亡，不是死于原发肿瘤，而是癌症转移或扩散到身体其他器官。据认为，一些细胞可以从原发或继发肿瘤中剥离并在血液中循环，或者在相对低氧的骨髓中找到安全的避难所，其中一些罕见的肿瘤细胞(CTCs和DTC)可以导致转移生长，特别是如果它们具有干细胞样的特征。最终目标是展示一种具有变革性的能力来捕获稀有细胞，特别是在用于捕获它们的同一平台上操作稀有细胞，并在同一芯片上单独处理液滴中捕获的细胞以进行分子表征。这项研究中体现的新想法是使用许多含有免疫磁珠的小液滴来探测稀有细胞，因为含有抗体的捕获珠以平行和流水线的方式顺序孵化和用液滴洗涤。在捕获稀有细胞方面，轻轻地用磁珠悬浮液探测小体积应该比探测大体积要有效得多，代价是需要许多重复(但快速)的步骤才能最终采集全部体积的样本。此外，可以使用多个抗体来探测一个样本。我们的目标是1)证明稀有细胞的亚群可以在单独的单个液滴的管道中捕获；以及2)通过实施旨在开发这些细胞的全部代谢潜力的分析，证明含有稀有细胞类型的液滴可以被操纵并输送到一个地点进行生化分析。

DESCRIPTION (provided by applicant): We are motivated by the view that development of a reliable and robust technology for efficient detection, characterization, and direct assay of minimal residual disease (MRD), including circulating tumor cells (CTCs) from blood and disseminated tumor cells (DTCs) from bone marrow, will make a major contribution toward elucidating the biology of the metastatic process and developing new methods for the management and treatment of cancer. These cells are important actors in how cancer may spread and kill people. Cancer is a dismal disease - within five years, almost 30% of cancer patients die, not from the primary tumor, but from the metastases or spread of the cancer to other organs in the body. It is thought that some cells can slough off the primary or secondary tumor and circulate in the bloodstream or find safe harbor in the relatively hypoxic bone marrow, and that some of these rare tumor cells (CTCs and DTCs) can lead to metastatic growth, especially if they acquire stem-cell like characteristics. The end goal is to demonstrate a transformative ability to capture and especially to manipulate rare cells on the same platform used to capture them, and to individually process captured cells in droplets for molecular characterization on the same chip. The novel idea embodied in this research is to use many small droplets containing immunomagnetic beads that probe for rare cells as the capture beads containing antibodies are sequentially incubated and washed with fluid droplets, in both a parallel and pipelined fashion. Gently probing small volumes with suspensions of magnetic beads should be much more efficient at capturing rare cells than probing a large volume, at the expense of requiring many repeated (but rapid) steps to eventually sample the full volume. Moreover, multiple antibodies may be used to probe one sample. Our aims are 1) to demonstrate that sub-populations of rare cells can be captured in a pipeline of separate, individual droplets; and 2) to show that droplets containing rare cell types can be manipulated and transported to a site for biochemical profiling, by implementing assays designed towards exploiting the full metabolic potential of these cells.