A new method for the detection of tumor single-cell drug sensitivity was developed by Qingdao Energy of the Chinese Academy of Sciences

January 18, reporters from China
Qingdao Institute of Bioeneress and Processes was informed that the institute's single-cell center successfully developed a new method based on the Raman group of tumor single-cell drug sensitivity detection D2O-CANST-R, with fast, low-cost, single-cell accuracy, identification of drug-resistant cells, embodiment of anti-cancer mechanisms, can be connected to single-cell sorting and sequencing and other characteristics, for cancer cell-drug mutual research, anti-cancer drug screening and other new means. The results of this research were published recently in Analytic Chemistry. The work was carried out under the auspices of Xu Jian Researcher of the Institute.
drug sensitivity detection method is the premise of cancer drug evaluation and screening, and also the basis of clinical chemotherapy program design.
chemotherapy play an important role in the treatment of malignant tumors, such as the right use, simple or complementary chemotherapy can treat some tumors, for some advanced tumors, chemotherapy can also be used for palliative care. However, there are significant differences in drug stress response between various tumor types or between individual patients, and the production of drug-resistant cells during chemotherapy can greatly reduce the efficacy of anticancer drugs. Therefore, fast, low-cost, drug-resistant cells can be identified, revealing the drug stress mechanism of tumor drug sensitivity detection methods, for cancer drug development and clinical precision drug use are essential.
According to the introduction, at present, the mainstream tumor drug sensitivity detection methods, such as color, bioluminescence, fluorescence analysis, etc., usually rely on the end-point test, that is, to distinguish cell death, but it is difficult to quantitatively and specifically measure the degree of "metabolic inhibition" of the drug to cancer cells. At the same time, based on the detection of cell population response, it is difficult to detect very few drug-resistant cells in cancer cell groups;
In response to this bottleneck, Qingdao Energy Institute single-cell center Maryam Hekmatara and others to human breast cancer cell strain (MCF-7) and rapamycin as an example of the development of heavy water feeding single-cell Raman spectral tumor drug sensitivity rapid detection technology (D2O-probed CANcer Susceptibility Test Ramanometry; D2O-CANST-R; Figure 1). Combined with tumor cell Raman group acquisition and multi-curve resolution-alternating least-multiplication analysis algorithm (MCR-ALS), the researchers demonstrated that after 1 to 3 days of drug treatment, D2O-CANST-R was able to specifically detect tumor drug sensitivity based on "metabolic inhibition" and to be able to detect tumor drug sensitivity based on "metabolic inhibition" and to be able to distinguish between cell nuclei, cell cytons, lipids and other individual cytoplers, tracking and distinguishing the rate and metabolic changes of proteins and lipids. The high activity of lipid and protein metabolism is an important reason for the rapid proliferation of tumor cells, so the above-mentioned ability is of great value for the mechanism research and screening of anti-cancer drugs.
Based on concepts such as "ramanome" and "drug stress Lamange" (Raman Barcode; RBCS) proposed earlier in the single-cell center, the researchers also revealed profound differences in single-cell precision metabolic stress mechanisms between etonucleotic organisms (human breast cancer cells and yeast cells), between cells, between drug concentrations, between drug processing time, and between biomolecule metabolic pathways. Therefore, D2O-CANST-R also has the characteristics of high space-time resolution, abundant information and revealing metabolic mechanism.
the method also found that after high doses of repamycin (500 or 5000×IC50) treatment, there are still cancer cells that maintain high metabolic activity, i.e. drug-resistant cells. D2O-CANST-R's ability to identify tumor-resistant cells and determine drug-resistant heterogeneity is of great significance for drug mechanism research, cancer drug evaluation and screening, and also has the potential to assist in the design of precision chemotherapy regimens.
The single-cell center of the Institute for clinical anti-infective drugs, put forward the "heavy water feeding single-cell Raman drug sensitivity test" principle, introduced the "minimum metabolic activity inhibition concentration" (MIC-MA) as a measure of drug sensitivity of the new concept, invented the "single-cell photo-light micro-drop Raman selection" (RAGE) and "single-cell micro-drop flow" Core devices such as Raman sequencing (RADS) have been successfully developed, such as the Clinical Single-Cell Raman Drug Sensitivity Fast Tester (CAST-R) and the Single-Cell Raman Sub-selection-Sequencing Coupling System (RACS-Seq), etc., and for clinical samples, the feasibility of simultaneous determination of antibiotic drug sensitivity ideographic and high-coverage genomes by individual bacterial cell accuracy has been demonstrated in clinical samples (Xu T, et, Small, 2020). This work is the above-mentioned single-celled technology system for human cells and drug interoperability expansion, will not only serve the research and development of tumor drugs, tumor precision drug use, but also for tumor single-celled selection and multi-group research to provide a new technical route.