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

描述（申请人提供）：我们的动机是，开发一种可靠和稳健的技术，用于有效检测、表征和直接测定微小残留病（MRD），包括来自血液的循环肿瘤细胞（CTC）和来自骨髓的播散性肿瘤细胞（DTC），将为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。这些细胞是癌症如何扩散和杀死人的重要因素。癌症是一种令人沮丧的疾病-在五年内，几乎30%的癌症患者不是死于原发性肿瘤，而是死于癌症转移或扩散到体内其他器官。人们认为，一些细胞可以从原发性或继发性肿瘤上斯劳，并在血液中循环，或者在相对缺氧的骨髓中找到安全港，并且其中一些罕见的肿瘤细胞（CTCs和DTCs）可能导致转移性生长，特别是如果它们获得干细胞样特征。最终目标是展示一种变革性的捕获能力，特别是在用于捕获稀有细胞的同一平台上操纵它们，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。在这项研究中体现的新思想是使用许多含有免疫磁珠的小液滴，这些小液滴探测稀有细胞，因为含有抗体的捕获珠以平行和流水线方式依次孵育并用流体液滴洗涤。用磁珠悬浮液轻轻探测小体积在捕获稀有细胞方面应该比探测大体积更有效，代价是需要许多重复的（但快速的）步骤来最终对全体积进行采样。此外，可以使用多种抗体来探测一个样品。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）通过实施旨在利用这些细胞的全部代谢潜力的测定，表明可以操纵含有稀有细胞类型的液滴并将其运输到用于生化分析的位点。

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.