High-flow microfluidics of leukapheresis blood products for functional analysis of breast circulating tumor cells

液体活组织检查有望为癌症治疗带来革命性的变化，因为它能够频繁地监测血液中的 肿瘤衍生材料，因为癌症对治疗干预的反应而演变。没有任何地方比这更 比三阴性转移性乳腺癌更明显，在转移性乳腺癌中，女性可能会经历多个系列的疗程 经过多年的治疗，每一次都与最初的反应有关，随后是药物的获得 抵抗。血浆DNA(CtDNA)突变分析提供了一种肿瘤负担的测量方法，并可能识别 小分子抑制剂的靶向突变，但它们不允许完整循环的体外培养 肿瘤细胞(CTCs)用于个体化的临床前药物敏感性(“精确肿瘤学”)。CTCS HAS的分离 使用小容量(5-10毫升)的细胞已经实现，但这些细胞非常罕见，因此不可能 建立临床可靠的体外CTC培养体系。我们的假设是，我们可以大量增加 血液通过标准的临床白细胞采集进行筛选(1-2名L)，作为回报，实现了100+倍的增长 建立CTCs常规体外培养所需的分离CTCs数量。同样重要的是要注意到，体外 通过采样1-2个L血获得的CTC培养将反映实时的分子亚型，遗传 单个患者转移性乳腺癌的组成和药物敏感性变化情况，包括 多个病变，共同构成血液传播的CTC群体和患者的整体肿瘤 负担。我们将开发一种高通量微流控技术，实现对 标记的血细胞远离未经处理的单个和成群的CTC，并提供与肿瘤无关的功能 “浓缩”性能，适用于从任何实体肿瘤类型播散的细胞，而不是 对肿瘤细胞的物理和/或生物学特性的任何先验假设。我们还将建立 转移性肿瘤患者体外高效CTC培养的微环境条件 三重阴性乳腺癌。成功将对新药开发和 在乳腺癌患者和其他患者的现有治疗方法中进行个性化的患者选择。

Liquid biopsies are poised to revolutionize cancer therapies, by enabling frequent blood-based monitoring of tumor-derived materials, as cancers evolve in response to therapeutic interventions. Nowhere is this more evident than in triple negative metastatic breast cancer, where women may undergo multiple serial courses of therapy over many years, each associated with an initial response, followed by the acquisition of drug resistance. Plasma DNA (ctDNA) mutation analyses provide a measure of tumor burden and may identify targetable mutations for small molecule inhibitors, but they do not allow ex vivo culture of intact circulating tumor cells (CTCs) for individualized preclinical drug sensitivity (“precision oncology”). Isolation of CTCs has been achieved using small blood volumes (5-10 mL), but these cells are so rare that it has not been feasible to establish clinically robust ex vivo CTC cultures. Our hypothesis is that we can massively increase the amount of blood screened through standard clinical leukapheresis (1-2 L), and in return, achieve 100+-fold increase in the number of isolated CTCs to establish routine ex vivo culture of CTCs. It is also important to note that ex vivo CTC cultures obtained by sampling 1-2 L blood will reflect the “real time” molecular subtype, genetic composition, and evolving drug sensitivity profile of an individual patient's metastatic breast cancer, including multiple lesions that together contribute to the blood-borne CTC population and the patient's overall tumor burden. We will develop a high-throughput microfluidic technology that achieves highly efficient depletion of tagged blood cells away from unmanipulated single and clustered CTCs and provides “tumor independent enrichment” performance, which is applicable to cells disseminated from any solid tumor type without making any a priori assumption on the physical and/or biological properties of tumor cells. We will also establish the microenvironmental conditions conducive to high efficiency ex vivo CTC cultures from women with metastatic triple negative breast cancer. Success would be transformative both for new drug development and for individualized patient selection among existing therapies for breast cancer patients, and others.