Journal Article

Determination of ADP/ATP translocase isoform ratios in malignancy and cellular senescence

Korandova, Zuzana, Kratky, Marek, Bartek, Jiri, Hodny, Zdenek, Tambor, Vojtech, Vasicova, Pavla, Maurencova, Dominika, Mracek, Tomas, Rysanek, David, Andera, Ladislav, Fabrik, Ivo, Kashmel, Pavel, Pecinova, Alena, Sultana, Pinky, Novak, Josef, Salovska, Barbora, Liblova, Zuzana, Kupcik, Rudolf, Vajrychova, Marie

2025

Overview

Cellular senescence has recently been recognized as a significant contributor to the poor prognosis of glioblastoma, one of the most aggressive brain tumors. Consequently, effectively eliminating senescent glioblastoma cells could benefit patients. Human ADP/ATP translocases (ANTs) play a role in oxidative phosphorylation in both normal and tumor cells. Previous research has shown that the sensitivity of senescent cells to mitochondria‐targeted senolytics depends on the level of ANT2. Here, we systematically mapped the transcript and protein levels of ANT isoforms in various types of senescence and glioblastoma tumorigenesis. We employed bioinformatics analysis, targeted mass spectrometry, RT‐PCR, immunoblotting, and assessment of cellular energy state to elucidate how individual ANT isoforms are expressed during the development of senescence in noncancerous and glioblastoma cells. We observed a consistent elevation of ANT1 protein levels across all tested senescence types, while ANT2 and ANT3 exhibited variable changes. Alterations in ANT protein isoform levels correlated with shifts in the cellular oxygen consumption rate. Our findings suggest that ANT isoforms are mutually interchangeable for oxidative phosphorylation and manipulating individual ANT isoforms could have potential for senolytic therapy. The individual functions of three isoforms exchanging ADP and ATP (ADP/ATP translocases; ANTs) on the mitochondrial membrane remain unclear. We developed a method for quantitatively differentiating highly similar human ANT1, ANT2, and ANT3 using parallel reaction monitoring. This method allowed us to assess changes in translocase levels during cellular senescence and correlate them with alterations in energy metabolism.

• Publisher: John Wiley & Sons, Inc,Wiley
• Subject: ADP/ATP translocase / Bioinformatics / Biosynthesis / Brain Neoplasms - enzymology / Brain Neoplasms - genetics / Brain Neoplasms - metabolism / Brain Neoplasms - pathology / Brain tumors / Cancer / Cell Line, Tumor / cellular senescence / Cellular Senescence - genetics / Chromatography / Cyclin-dependent kinases / Ethylenediaminetetraacetic acid / Fibroblasts / Genotype & phenotype / Glioblastoma / Glioblastoma - enzymology / Glioblastoma - genetics / Glioblastoma - metabolism / Glioblastoma - pathology / Glioblastoma cells / Glioma / Gliomas / Glucose / Humans / Immunoblotting / Isoenzymes - genetics / Isoenzymes - metabolism / Isoforms / Kinases / Malignancy / Mass spectrometry / Mass spectroscopy / Metabolism / Mitochondrial ADP, ATP Translocases - genetics / Mitochondrial ADP, ATP Translocases - metabolism / Oxidative Phosphorylation / Oxygen consumption / Penicillin / Phosphorylation / Physiology / Prognosis / Protein Isoforms - genetics / Protein Isoforms - metabolism / Proteins / Radiation / Scientific imaging / Senescence / targeted mass spectrometry / Telomerase / Tetracycline / Tetracyclines / Translocase / Tumor cells / Tumorigenesis