Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action

Abstract

Studying the mechanisms of drug antitumor activity at the single-cell level can provide information about the responses of cell subpopulations to drug therapy, which is essential for the accurate treatment of cancer. Due to the small size of single cells and the low contents of metabolites, metabolomics-based approaches to studying the mechanisms of drug action at the single-cell level are lacking. Herein, we develop a label-free platform for studying the mechanisms of drug action based on single-cell metabolomics (sMDA-scM) by integrating intact living-cell electro-launching ionization mass spectrometry (ILCEI-MS) with metabolomics analysis. Using this platform, we reveal that non-small-cell lung cancer (NSCLC) cells treated by gefitinib can be clustered into two cell subpopulations with different metabolic responses. The glutathione metabolic pathway of the subpopulation

Exploring the Accumulation Behavior and Heterogeneity of Perfluorooctanesulfonic Acid in Zebrafish Primary Organ Cells by Single-Cell Mass Cytometry

“…capillary, forming a single-cell flow (Figure S3). The cells intercepted at the outlet of the etched capillary were in a monodisperse manner with well-preserved cellular structures (Figure S4), suggesting the ionization mechanism of our single-cell mass cytometry method was quite similar to that of the ILCEI-MS method, 26,27 of which an intact cell entered into the inlet of mass spectrometer and further ionized in the iontransfer tube for mass spectrometric analysis. ILCEI-MS used a narrow-bore capillary with constant-I.D.…”

“…of 16 μm to achieve single-cell separation and transport, and nitrogen was needed to drive the single cells into the capillary in a high-pressure chamber. 26,27 In our study, a capillary with constant-I.D. of 25 μm (slightly larger than the average cell diameter) was used for single-cell transportation, and only a peristaltic pump was enough to introduce cell suspensions; thus, the configuration of the single-cell mass cytometry system was greatly simplified.…”

“…To overcome this problem, single-cell mass cytometry, which combines the high-throughput feature of flow cytometry and the simultaneous acquisition of multiplexed information on organic MS, has developed rapidly in recent years. Mass cytometry by ESI-MS, , nanoESI-MS, dielectric barrier discharge ionization (DBDI)-MS, and intact living-cell electrolaunching ionization (ILCEI)-MS , have been developed as new choices for in-depth characterization of heterogeneous cell populations to interrogate multiple levels of cellular metabolism.…”

Single-Cell Unsaturated Lipid Profiling for Studying Chemoresistance Heterogeneity of Triple-Negative Breast Cancer Cells

“…This platform has been used to elucidate the molecular mechanisms based on the metabolic characteristics of cancer cells. For example, based on CyESI-MS, the metabolic changes of liver cancer cells after being cocultured with natural killer (NK) cells revealed the metabolic pathway regulation of cancer cells in resisting NK immunity. , Other research has also employed a similar MS flow cytometry platform for revealing the metabolic mechanism of drug resistance in nonsmall cell lung cancer cells …”

Supramolecular Anchored Copper Nanoclusters for a Multipath Electrochemiluminescence Probe

“…Utilizing small-molecule ligands to stabilize Cu NCs can effectively diminish the thickness of the insulating layer present on their surface, thereby enhancing the efficiency of carrier transport and, ultimately, augmenting their ECL performance. 18,19 Regrettably, Cu NCs protected by smallmolecule ligands generally suffer from inadequate solution stability and a propensity to undergo aggregation and stacking phenomena. To obtain stable, well-dispersed, and high ECL efficiency Cu NCs still remains a challenge in the field of ECL.…”