Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator.
Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator.
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DOI：
10.1371/journal.pone.0264907
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
Hur SC
中科院分区：
文献类型：
--
作者：
Sung HW;Choi SE;Chu CH;Ouyang M;Kalyan S;Scott N;Hur SC
患者样本的直接评估在癌症治疗中具有前所未有的潜力。液体活检中的循环肿瘤细胞（CTC）是临床上原代细胞的快速发展来源，并且是功能测定的理想候选者，以实时揭示实时肿瘤信息。然而，缺乏允许直接从液体活检样品直接和主动询问CTC的常规代表了液体活检在临床环境中的转化使用的瓶颈。为了解决这个问题，我们提出了一个工作流程，使用微流控涡旋辅助电穿孔系统，设计用于从血液中纯化的CTC的功能评估。通过对野生型（HCC 827 wt）和吉非替尼耐药（HCC 827 GR6）非小细胞肺癌（NSCLC）细胞的药物反应试验评估了该方法的验证。将捕获在微尺度涡旋内的HCC 827细胞电穿孔以将药物试剂依次递送到胞质溶胶中。使用自动单细胞图像荧光强度算法，对两种细胞系表征促进多药剂递送的电穿孔条件。将HCC827 GR6细胞加标到血液中以模拟耐药CTC，能够以高纯度收集，证明该装置能够最大限度地减少背景细胞对下游敏感细胞测定的影响。使用我们提出的工作流程，恢复吉非替尼敏感性的药物组合反映了预期的细胞毒性反应。总之，这些结果代表了微流体多药物筛选小组的工作流程，其可以实现原位对患者CTC的功能询问，从而加速液体活检的临床标准化。
Direct assessment of patient samples holds unprecedented potential in the treatment of cancer. Circulating tumor cells (CTCs) in liquid biopsies are a rapidly evolving source of primary cells in the clinic and are ideal candidates for functional assays to uncover real-time tumor information in real-time. However, a lack of routines allowing direct and active interrogation of CTCs directly from liquid biopsy samples represents a bottleneck for the translational use of liquid biopsies in clinical settings. To address this, we present a workflow for using a microfluidic vortex-assisted electroporation system designed for the functional assessment of CTCs purified from blood. Validation of this approach was assessed through drug response assays on wild-type (HCC827 wt) and gefitinib-resistant (HCC827 GR6) non-small cell lung cancer (NSCLC) cells. HCC827 cells trapped within microscale vortices were electroporated to sequentially deliver drug agents into the cytosol. Electroporation conditions facilitating multi-agent delivery were characterized for both cell lines using an automatic single-cell image fluorescence intensity algorithm. HCC827 GR6 cells spiked into the blood to emulate drug-resistant CTCs were able to be collected with high purity, demonstrating the ability of the device to minimize background cell impact for downstream sensitive cell assays. Using our proposed workflow, drug agent combinations to restore gefitinib sensitivity reflected the anticipated cytotoxic response. Taken together, these results represent a microfluidics multi-drug screening panel workflow that can enable functional interrogation of patient CTCs in situ, thereby accelerating the clinical standardization of liquid biopsies.
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