Research output: Contribution to journal › Article › peer-review
Resistance to H2O2-induced oxidative stress in human cells of different phenotypes. / Zenin, Valeriy; Ivanova, Julia; Pugovkina, Natalia; Shatrova, Alla; Aksenov, Nikolay; Tyuryaeva, Irina; Kirpichnikova, Kseniya; Kuneev, Ivan; Zhuravlev, Andrei; Osyaeva, Ekaterina; Lyublinskaya, Ekaterina; Gazizova, Ilyuza; Guriev, Nikita; Lyublinskaya, Olga.
In: Redox Biology, Vol. 50, No. 102245, 102245, 01.04.2022.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Resistance to H2O2-induced oxidative stress in human cells of different phenotypes
AU - Zenin, Valeriy
AU - Ivanova, Julia
AU - Pugovkina, Natalia
AU - Shatrova, Alla
AU - Aksenov, Nikolay
AU - Tyuryaeva, Irina
AU - Kirpichnikova, Kseniya
AU - Kuneev, Ivan
AU - Zhuravlev, Andrei
AU - Osyaeva, Ekaterina
AU - Lyublinskaya, Ekaterina
AU - Gazizova, Ilyuza
AU - Guriev, Nikita
AU - Lyublinskaya, Olga
N1 - Publisher Copyright: © 2022 The Authors
PY - 2022/4/1
Y1 - 2022/4/1
N2 - Application of genetically encoded biosensors of redox-active compounds promotes the elaboration of new methods for investigation of intracellular redox activities. Previously, we have developed a method to measure quantitatively the intracellular concentration of hydrogen peroxide (H2O2) in living cells using genetically encoded biosensor HyPer. In the present study, we refined the method and applied it for comparing the antioxidant system potency in human cells of different phenotypes by measuring the gradient between the extracellular and cytoplasmic H2O2 concentrations under conditions of H2O2-induced external oxidative stress. The measurements were performed using cancer cell lines (K-562 and HeLa), as well as normal human cells – all expressing HyPer in the cell cytoplasm. As normal cells, we used three isogenic lines of different phenotypes – mesenchymal stem/stromal cells (MSCs), induced pluripotent stem cells (iPSCs) derived from MSCs by reprogramming, and differentiated iPSC progenies with the phenotype resembling precursory MSCs. When exposing cells to exogenous H2O2, we showed that at low oxidative loads (<50 μM of H2O2) the gradient depended on extracellular H2O2 concentration. At high loads (>50 μM of H2O2), which caused the exhaustion of thioredoxin activity in the cell cytoplasm, the gradient stabilized, pointing out that it is the functional status of the thioredoxin-depended enzymatic system that drives the dependence of the H2O2 gradient on the oxidative load in human cells. At high H2O2 concentrations, the cytoplasmic H2O2 level in cancer cells was found to be several hundred times lower than the extracellular one. At the same time, in normal cells, extracellular-to-intracellular gradient amounted to thousands of times. Upon reprogramming, the potency of cellular antioxidant defense increased. In contrast, differentiation of iPSCs did not result in the changes in antioxidant system activity in the cell cytoplasm, assuming that intensification of the H2O2-detoxification processes is inherent to a period of early human development.
AB - Application of genetically encoded biosensors of redox-active compounds promotes the elaboration of new methods for investigation of intracellular redox activities. Previously, we have developed a method to measure quantitatively the intracellular concentration of hydrogen peroxide (H2O2) in living cells using genetically encoded biosensor HyPer. In the present study, we refined the method and applied it for comparing the antioxidant system potency in human cells of different phenotypes by measuring the gradient between the extracellular and cytoplasmic H2O2 concentrations under conditions of H2O2-induced external oxidative stress. The measurements were performed using cancer cell lines (K-562 and HeLa), as well as normal human cells – all expressing HyPer in the cell cytoplasm. As normal cells, we used three isogenic lines of different phenotypes – mesenchymal stem/stromal cells (MSCs), induced pluripotent stem cells (iPSCs) derived from MSCs by reprogramming, and differentiated iPSC progenies with the phenotype resembling precursory MSCs. When exposing cells to exogenous H2O2, we showed that at low oxidative loads (<50 μM of H2O2) the gradient depended on extracellular H2O2 concentration. At high loads (>50 μM of H2O2), which caused the exhaustion of thioredoxin activity in the cell cytoplasm, the gradient stabilized, pointing out that it is the functional status of the thioredoxin-depended enzymatic system that drives the dependence of the H2O2 gradient on the oxidative load in human cells. At high H2O2 concentrations, the cytoplasmic H2O2 level in cancer cells was found to be several hundred times lower than the extracellular one. At the same time, in normal cells, extracellular-to-intracellular gradient amounted to thousands of times. Upon reprogramming, the potency of cellular antioxidant defense increased. In contrast, differentiation of iPSCs did not result in the changes in antioxidant system activity in the cell cytoplasm, assuming that intensification of the H2O2-detoxification processes is inherent to a period of early human development.
KW - Cell reprogramming
KW - Genetically encoded biosensors
KW - HO
KW - HO gradient
KW - Hydrogen peroxide
KW - HyPer
KW - Induced pluripotent stem cells
KW - Kinetics
KW - Mesenchymal stem/stromal cells
KW - Rate constants
KW - Oxidative Stress
KW - Hydrogen Peroxide/metabolism
KW - Humans
KW - Mesenchymal Stem Cells/metabolism
KW - Phenotype
KW - HeLa Cells
KW - H2O2
KW - H2O2 gradient
UR - http://www.scopus.com/inward/record.url?scp=85123742039&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/a42ea648-00c1-352d-9178-2ed0afcbc5dd/
U2 - 10.1016/j.redox.2022.102245
DO - 10.1016/j.redox.2022.102245
M3 - Article
C2 - 35114579
AN - SCOPUS:85123742039
VL - 50
JO - Redox Biology
JF - Redox Biology
SN - 2213-2317
IS - 102245
M1 - 102245
ER -
ID: 98153940