Jiang Ruoyu, Agrawal Sudhanshu, Aghaamoo Mohammad, Parajuli Ritesh, Agrawal Anshu, Lee Abraham P
Biomedical Engineering, University of California, Irvine, CA 92697, USA.
Department of Medicine, Division of Basic and Clinical Immunology, University of California, Irvine, CA 92697, USA.
Lab Chip. 2021 Mar 9;21(5):875-887. doi: 10.1039/d0lc00969e.
We demonstrate a label free and high-throughput microbubble-based acoustic microstreaming technique to isolate rare circulating cells such as circulating cancer associated fibroblasts (cCAFs) in addition to circulating tumor cells (CTCs) and immune cells (i.e. leukocytes) from clinically diagnosed patients with a capture efficiency of 94% while preserving cell functional integrity within 8 minutes. The microfluidic device is self-pumping and was optimized to increase flow rate and achieve near perfect capturing of rare cells enabled by having a trapping capacity above the acoustic vortex saturation concentration threshold. Our approach enables rapid isolation of CTCs, cCAFs and their associated clusters from blood samples of cancer patients at different stages. By examining the combined role of cCAFs and CTCs in early cancer onset and metastasis progression, the device accurately diagnoses both cancer and the metastatic propensity of breast cancer patients. This was confirmed by flow cytometry where we observed that metastatic breast cancer blood samples had significantly higher percentage of exhausted CD8+ T cells expressing programmed cell death protein 1 (PD1), higher number of CD4+ T regulatory cells and T helper cells. We show for the first time that our lateral cavity acoustic transducers (LCATs)-based approach can thus be developed into a metastatic propensity assay for clinical usage by elucidating cancer immunological responses and the complex relationships between CTCs and its companion tumor microenvironment.
我们展示了一种基于微泡的无标记高通量声流技术，可从临床诊断患者中分离出罕见的循环细胞，如循环肿瘤相关成纤维细胞（cCAF）、循环肿瘤细胞（CTC）和免疫细胞（即白细胞），捕获效率达94%，并在8分钟内保持细胞功能完整性。该微流控装置具有自泵功能，并经过优化以提高流速，通过使捕获能力高于声涡饱和浓度阈值，实现对罕见细胞的近乎完美捕获。我们的方法能够从不同阶段癌症患者的血液样本中快速分离出CTC、cCAF及其相关簇。通过研究cCAF和CTC在癌症早期发病和转移进展中的联合作用，该装置可准确诊断乳腺癌患者的癌症及转移倾向。流式细胞术证实了这一点，我们观察到转移性乳腺癌血液样本中表达程序性细胞死亡蛋白1（PD1）的耗竭CD8 + T细胞百分比显著更高，CD4 + T调节细胞和辅助性T细胞数量更多。我们首次表明，基于侧向腔声换能器（LCAT）的方法通过阐明癌症免疫反应以及CTC与其伴随的肿瘤微环境之间的复杂关系，可发展成为一种用于临床的转移倾向检测方法。
