A high-throughput assay for screening natural products that boost NK cell-mediated killing of cancer cells

Abstract

Context: Natural killer (NK) cells can eliminate malignant cells and play a vital role in immunosurveillance. Administration of natural compounds represents a promising approach for antitumor immunotherapy, which may enhance the NK cell activity via multiple mechanisms. Objective: Establishing approaches to evaluate the effect of select natural products on NK cell-mediated cytotoxicity. Materials and methods: We selected a natural product library containing 2880 pure compounds, which was provided by the National Centre for Drug Screening of China. 0.1% DMSO was employed as a negative control, and 100 U/mL human recombinant IL-2 was employed as a positive control. To evaluate the % of tumour cells which were killed by NK cells, expanded NK cells were co-cultured with tumour cells and then treated with natural products at the concentration of 10 lM. After 24-h co-incubation, luminescent signal was detected and percent lysis was calculated. Results: We report on the results of a three-round high-throughput screening effort that identified 20deoxyingenol 3-angelate (DI3A) and its analogue ingenol 3-angelate (I3A) as immuno enhancers which boosts NK cell-mediated killing of non-small cell lung cancer cells (NSCLCs). Biophotonic cytotoxicity assay and calcein release assay were used as two well-established NK cell cytotoxicity detection assays to validate the immuno-enhancing effects of DI3A and I3A, which was achieved by increasing degranulation and interferon-gamma secretion of NK cells. Conclusions: Our newly established ATP-based method was a valuable and information-rich screening tool to investigate the biological effects of natural products on both NK cells and tumour cells.

Targeting CD155 by rediocide-A overcomes tumour immuno-resistance to natural killer cells

“…Previously, we screened nearly 3000 natural products using a high-throughput assay (Gong et al 2015;Xu et al 2020) and found that Red-A increased NK cell-mediated killing of NSCLCs. We demonstrated Red-A could affect the viabilities of A549 and H1299 tumour cells but did not affect the viabilities of NK cells (Figure 1(A-C)).…”

“…In search of a natural substance that overcomes tumour immune resistance to NK cells, we performed a high-throughput assay to screen nearly 3000 Chinese medicine-derived small molecular compounds in our previous study (Gong et al 2015;Xu et al 2020). We found that rediocide-A (Red-A) extracted from the roots of Trigonostemon rediocides Craib (Euphorbiaceae) could potentially block tumour immune resistance to NK cells.…”

Thevebioside, the Active Ingredient of Traditional Chinese Medicine, Inhibits SRC3 Mediated Signaling by Ubiquitination to Induce NSCLC Cells Apoptosis

“…In our previous studies, we established a new luminescence-based high-throughput screening system [11], which could detect percent lysis of tumor cells and its viability with natural products treatment ( Fig. 1A).…”

“…In our previous study, we have established a new type of high-throughput screening system to search for natural products with anti-tumor activities [11]. THB is one of the natural product candidates that we used this luminescence-based screening system to nd out, which sensitive to NSCLC cells.…”

Role of Plant-Derived Natural Compounds in Experimental Autoimmune Encephalomyelitis: A Review of the Treatment Potential and Development Strategy

“…There have been many studies of the treatment of EAE by TCM monomers containing flavonoids, phenol, and glycosides, among others ( Ciumărnean et al, 2020 ; Ikram et al, 2020 ; Yang L. et al, 2020 ). Moreover, many researchers have studied the effect of chemical monomers by molecular docking, high-throughput screening, and network pharmacology ( Chen et al, 2020 ; Liu J. et al, 2020 ; Xu et al, 2020a ). Over the past 8 years, glycosides have been the most commonly studied compounds, and have been examined and developed with the intent of using them as a potential treatment for EAE ( Giacoppo et al, 2013 ; Yin et al, 2014 ; Haghmorad et al, 2017 ; Madhu et al, 2019 ; Yang L. et al, 2020 ).…”