Microfluidic Single-Cell Proteomics Assay Chip: Lung Cancer Cell Line Case Study.

Yugyung Jung,Yu Ri Nam,James R Heath,Jongchan Choi,Sung Yang,Minkook Son

doi:10.3390/mi12101147

Abstract

Cancer is a dynamic disease involving constant changes. With these changes, cancer cells become heterogeneous, resulting in varying sensitivity to chemotherapy. The heterogeneity of cancer cells plays a key role in chemotherapy resistance and cancer recurrence. Therefore, for effective treatment, cancer cells need to be analyzed at the single-cell level by monitoring various proteins and investigating their heterogeneity. We propose a microfluidic chip for a single-cell proteomics assay that is capable of analyzing complex cellular signaling systems to reveal the heterogeneity of cancer cells. The single-cell assay chip comprises (i) microchambers (n = 1376) for manipulating single cancer cells, (ii) micropumps for rapid single-cell lysis, and (iii) barcode immunosensors for detecting nine different secretory and intracellular proteins to reveal the correlation among cancer-related proteins. Using this chip, the single-cell proteomics of a lung cancer cell line, which may be easily masked in bulk analysis, were evaluated. By comparing changes in the level of protein secretion and heterogeneity in response to combinations of four anti-cancer drugs, this study suggests a new method for selecting the best combination of anti-cancer drugs. Subsequent preclinical and clinical trials should enable this platform to become applicable for patient-customized therapies.

Highlights

Cancer is a dynamic disease that becomes increasingly heterogeneous as it progresses [1,2,3]
We propose a single-cell proteomics assay, comprising a microfluidic platform that can analyze cancer signal transduction systems to elucidate the heterogeneity of cancer cells
Of the 1376 chambers, 98.9% (±0.15%) acquired cancer cells, either in the form of single cells or multiple cells, and 89.5% (±3.7%) of the trapped cells were verified as single cells (Figure 2A)

Summary

Introduction

Cancer is a dynamic disease that becomes increasingly heterogeneous as it progresses [1,2,3]. Even after becoming malignant, and constantly change by acquiring a variety of mutations [2] With these continuous changes, cancer cells produce bulk tumors formed from groups of heterogeneous cells, which have various sensitivities to chemotherapy [3]. It was shown that intra-tumor heterogeneity can be used as one of the clinically important prognostic factors [8,9,10] This implies that patients with tumors in which the heterogeneity is high are more likely to have a poor prognosis for chemotherapy and a higher resistance to targeted treatment [11]. The most commonly used method for chemotherapy-resistant tumor cells is a combination of two or more anti-cancer drugs, potentially lowering the incidence of various drug-tolerant cancer cell groups due to the heterogeneity of the tumor [6]; for such strategies to be successful for Micromachines 2021, 12, 1147.

Methods

Results

Discussion

Conclusion