Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells

Aleksandr Klepinin; Sten Miller; Indrek Reile; Marju Puurand; Egle Rebane-Klemm; Ljudmila Klepinina; Heiki Vija; Song Zhang; Andre Terzic; Petras Dzeja; Tuuli Kaambre

doi:10.3389/fonc.2022.892195

Stable Isotope Tracing Uncovers Reduced γ/β-ATP Turnover and Metabolic Flux Through Mitochondrial-Linked Phosphotransfer Circuits in Aggressive Breast Cancer Cells

Front Oncol. 2022 May 31:12:892195. doi: 10.3389/fonc.2022.892195. eCollection 2022.

Authors

Aleksandr Klepinin^{1

2}, Sten Miller^{1

3}, Indrek Reile⁴, Marju Puurand¹, Egle Rebane-Klemm^{1

3}, Ljudmila Klepinina^{1

3}, Heiki Vija⁵, Song Zhang², Andre Terzic^{6

7}, Petras Dzeja², Tuuli Kaambre¹

Affiliations

¹ Laboratory of Chemical Biology, National Institute of Chemical Physics and Biophysics, Tallinn, Estonia.
² Department of Cardiovascular Medicine and Center for Regenerative Medicine, Mayo Clinic, Rochester, MN, United States.
³ Department of Chemistry and Biotechnology, School of Science, Tallinn University of Technology, Tallinn, Estonia.
⁴ Laboratory of Chemical Physics, National Institute of Chemical Physics and Biophysics, Tallinn, Estonia.
⁵ Laboratory of Environmental Toxicology, National Institute of Chemical Physics and Biophysics, Tallinn, Estonia.
⁶ Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN, United States.
⁷ Department of Clinical Genomics, Mayo Clinic, Rochester, MN, United States.

Abstract

Changes in dynamics of ATP γ- and β-phosphoryl turnover and metabolic flux through phosphotransfer pathways in cancer cells are still unknown. Using ¹⁸O phosphometabolite tagging technology, we have discovered phosphotransfer dynamics in three breast cancer cell lines: MCF7 (non-aggressive), MDA-MB-231 (aggressive), and MCF10A (control). Contrary to high intracellular ATP levels, the ¹⁸O labeling method revealed a decreased γ- and β-ATP turnover in both breast cancer cells, compared to control. Lower β-ATP[¹⁸O] turnover indicates decreased adenylate kinase (AK) flux. Aggressive cancer cells had also reduced fluxes through hexokinase (HK) G-6-P[¹⁸O], creatine kinase (CK) [CrP[¹⁸O], and mitochondrial G-3-P[¹⁸O] substrate shuttle. Decreased CK metabolic flux was linked to the downregulation of mitochondrial MTCK1A in breast cancer cells. Despite the decreased overall phosphoryl flux, overexpression of HK2, AK2, and AK6 isoforms within cell compartments could promote aggressive breast cancer growth.

Keywords: 18 O stable isotope labeling technology; adenylate kinase; creatine kinase; glycolysis; oxidative phosphorylation; phosphotransfer network; triple-negative breast cancer; γ-and β-ATP phosphoryl turnover.