Diverse temporal and spatial mechanisms work, partially through Stanniocalcin-1, V-ATPase and senescence, to activate the extracellular ATP-mediated drug resistance in human cancer cells

Haiyun Zhang; Jingwen Song; Ryan Ward; Yong Han; Arabella Hunt; Pratik Shriwas; Alexander Steed; Cory Edwards; Yanyang Cao; Milo Co; Xiaozhuo Chen

doi:10.3389/fonc.2024.1276092

Diverse temporal and spatial mechanisms work, partially through Stanniocalcin-1, V-ATPase and senescence, to activate the extracellular ATP-mediated drug resistance in human cancer cells

Front Oncol. 2024 Feb 6:14:1276092. doi: 10.3389/fonc.2024.1276092. eCollection 2024.

Authors

Haiyun Zhang^{1

2

3}, Jingwen Song^{1

2

3}, Ryan Ward⁴, Yong Han², Arabella Hunt⁴, Pratik Shriwas^{1

2

3}, Alexander Steed⁵, Cory Edwards⁵, Yanyang Cao^{1

2

3}, Milo Co⁵, Xiaozhuo Chen^{1

2

3

5

6}

Affiliations

¹ Department of Biological Science, Ohio University, Athens, OH, United States.
² The Edison Biotechnology Institute, Ohio University, Athens, OH, United States.
³ The Program of Molecular and Cellular Biology, Ohio University, Athens, OH, United States.
⁴ The Honor Tutorial College, Ohio University, Athens, OH, United States.
⁵ Heritage College of Osteopathic Medicine, Ohio University, Athens, OH, United States.
⁶ Department of Biomedical Science, Heritage College of Osteopathic Medicine, Ohio University, Athens, OH, United States.

Abstract

Introduction: Resistance to drug therapies is associated with a large majority of cancer-related deaths. ATP-binding cassette (ABC) transporter-mediated drug efflux, epithelial-mesenchymal transition (EMT), cancer stem cells (CSCs), glutathione (GSH), senescence, and vacuole-type ATPase (V-ATPase) all contribute to the resistance. We recently showed that extracellular ATP (eATP) induces and regulates EMT, CSC formation, and ABC transporters in human cancer cells and tumors. eATP also consistently upregulates Stanniocalcin-1 (STC1), a gene that significantly contributes to EMT, CSC formation, and tumor growth. We also found that eATP enhances drug resistance in cancer cells through eATP internalization mediated by macropinocytosis, leading to an elevation of intracellular ATP (iATP) levels, induction of EMT, and CSC formation. However, these factors have never been systematically investigated in the context of eATP-induced drug resistance.

Methods: In this study, we hypothesized that eATP increases drug resistance via inducing ABC efflux, EMT, CSCs, STC1, and their accompanied processes such as GSH reducing activity, senescence, and V-ATPase. RNA sequencing, metabolomics, gene knockdown and knockout, and functional assays were performed to investigate these pathways and processes.

Results and discussion: Our study results showed that, in multiple human cancer lines, eATP induced genes involved in drug resistance, elevated ABC transporters' efflux activity of anticancer drugs; generated transcriptomic and metabolic profiles representing a drug resistant state; upregulated activities of GSH, senescence, and V-ATPase to promote drug resistance. Collectively, these newly found players shed light on the mechanisms of eATP-induced as well as STC1- and V-ATPase-mediated drug resistance and offer potential novel targets for combating drug resistance in cancers.

Keywords: EMT; cancer metabolism; cancer stem cells; gene knockdown; lysosome; metabolomics; transcriptomics.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study is supported by an NIH grant R15CA242177 to XC and an Ohio University Student Enhancement Award to HZ, and John Kopchick MCB/TBS Undergraduate Student Research Fund, and Provost Undergraduate Research Fund to RW.