Suppression of Asparagine Synthetase Enhances the Antitumor Potency of ART and Artemalogue SOMCL-14-221 in NSCLC Cells

Abstract

Background: Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related mortality. Artemisinin (ART) and SOMCL-14-221 (221), a spirobicyclic analogue of ART, have been reported to inhibit the proliferation of A549 cells; yet, the underlying mechanism remains unclear. Methods: Both in vitro and in vivo experiments were performed to examine the anti-proliferative effects of ART and 221. The TMT-based quantitative proteomics and NMR-based metabolomics were conducted to analyze the global proteomic and metabolomic changes in ART and 221 groups, respectively. The potential mechanism of ART and 221 in suppressing tumor growth was studied by knockdown of asparagine synthetase (ASNS) and inhibition of ER stress. Findings: Both ART and 221 inhibited the proliferation of NSCLC cells in vitro and in vivo. The proteomic data revealed 114 upregulated proteins in ART and 221 groups, in which biological process of “response to endoplasmic reticulum stress” were highly represented. ASNS was identified as a key node protein in this process. Interestingly, knockdown of ASNS improved the cytotoxicity of ART and 221, and administration with ART and 221 disordered amino acid metabolism. Moreover, ART and 221 activated ER stress, and inhibition of ER stress abolished the anti-proliferative effects of ART and 221. Interpretation: This study demonstrates that ART and 221 suppress tumor growth by triggering ER stress, and the inhibition of ASNS enhances the antitumor activity of ART and 221, which provides new strategy of combined drug therapy. Funding Statement: This work was supported by the National Research and from the Ministry of Science and Technology of China (No. 2017YFC1700200), the National Science & Technology Major Project Key New Drug Creation and Manufacturing Program of China (Grant No. 2018ZX09711002), National Natural Science Foundation of China (No. 31800693, No. 21877120, 81430080) and the ''Personalized Medicines-Molecular Signature-based Drug Discovery and Development of Strategic Priority Research of the Chinese Academy of Sciences (No. XDA12020320). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: All animal studies (including the mice euthanasia procedure) were done in compliance with the regulations and guidelines of Shanghai Institute of Materia Medica (Chinese Academy of Sciences) institutional animal care and conducted according to the AAALAC and the IACUC guidelines.