Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion
Cooling at 4°C is routinely used to lower metabolism and preserve cell and tissue integrity in laboratory and clinical settings, including organ transplantation. However, cooling and rewarming produce cell damage, attributed primarily to a burst of reactive oxygen species (ROS) upon rewarming. While...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2022-07-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.1177/09636897221108705 |
| _version_ | 1811322810775109632 |
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| author | Marziyeh Tolouee Koen D. W. Hendriks Fia Fia Lie Lucas P. Gartzke Maaike Goris Femke Hoogstra-Berends Steven Bergink Robert H. Henning |
| author_facet | Marziyeh Tolouee Koen D. W. Hendriks Fia Fia Lie Lucas P. Gartzke Maaike Goris Femke Hoogstra-Berends Steven Bergink Robert H. Henning |
| author_sort | Marziyeh Tolouee |
| collection | DOAJ |
| description | Cooling at 4°C is routinely used to lower metabolism and preserve cell and tissue integrity in laboratory and clinical settings, including organ transplantation. However, cooling and rewarming produce cell damage, attributed primarily to a burst of reactive oxygen species (ROS) upon rewarming. While DNA represents a highly vulnerable target of ROS, it is unknown whether cooling and/or rewarming produces DNA damage. Here, we show that cooling alone suffices to produce extensive DNA damage in cultured primary cells and cell lines, including double-strand breaks (DSBs), as shown by comet assay and pulsed-field gel electrophoresis. Cooling-induced DSB formation is time- and temperature-dependent and coincides with an excess production of ROS, rather than a decrease in ATP levels. Immunohistochemistry confirmed that DNA damage activates the DNA damage response marked by the formation of nuclear foci of proteins involved in DSB repair, γ-H2Ax, and 53BP1. Subsequent rewarming for 24 h fails to recover ATP levels and only marginally lowers DSB amounts and nuclear foci. Precluding ROS formation by dopamine and the hydroxychromanol, Sul-121, dose-dependently reduces DSBs. Finally, a standard clinical kidney transplant procedure, using cold static storage in UW preservation solution up to 24 h in porcine kidney, lowered ATP, increased ROS, and produced increasing amounts of DSBs with recruitment of 53BP1. Given that DNA repair is erroneous by nature, cooling-inflicted DNA damage may affect cell survival, proliferation, and genomic stability, significantly impacting cellular and organ function, with relevance in stem cell and transplantation procedures. |
| first_indexed | 2024-04-13T13:41:41Z |
| format | Article |
| id | doaj.art-c756303882ec4058bd4fb4dc35d03866 |
| institution | Directory Open Access Journal |
| issn | 1555-3892 |
| language | English |
| last_indexed | 2024-04-13T13:41:41Z |
| publishDate | 2022-07-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-c756303882ec4058bd4fb4dc35d038662022-12-22T02:44:37ZengSAGE PublishingCell Transplantation1555-38922022-07-013110.1177/09636897221108705Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP DepletionMarziyeh Tolouee0Koen D. W. Hendriks1Fia Fia Lie2Lucas P. Gartzke3Maaike Goris4Femke Hoogstra-Berends5Steven Bergink6Robert H. Henning7Department of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsDepartment of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsDepartment of Pharmacology, Medical Faculty, Universitas Tarumanagara, Jakarta, IndonesiaDepartment of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsDepartment of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsDepartment of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsDepartment of Biomedical Sciences of Cells and Systems (BSCS), University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsDepartment of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, University of Groningen, Groningen, The NetherlandsCooling at 4°C is routinely used to lower metabolism and preserve cell and tissue integrity in laboratory and clinical settings, including organ transplantation. However, cooling and rewarming produce cell damage, attributed primarily to a burst of reactive oxygen species (ROS) upon rewarming. While DNA represents a highly vulnerable target of ROS, it is unknown whether cooling and/or rewarming produces DNA damage. Here, we show that cooling alone suffices to produce extensive DNA damage in cultured primary cells and cell lines, including double-strand breaks (DSBs), as shown by comet assay and pulsed-field gel electrophoresis. Cooling-induced DSB formation is time- and temperature-dependent and coincides with an excess production of ROS, rather than a decrease in ATP levels. Immunohistochemistry confirmed that DNA damage activates the DNA damage response marked by the formation of nuclear foci of proteins involved in DSB repair, γ-H2Ax, and 53BP1. Subsequent rewarming for 24 h fails to recover ATP levels and only marginally lowers DSB amounts and nuclear foci. Precluding ROS formation by dopamine and the hydroxychromanol, Sul-121, dose-dependently reduces DSBs. Finally, a standard clinical kidney transplant procedure, using cold static storage in UW preservation solution up to 24 h in porcine kidney, lowered ATP, increased ROS, and produced increasing amounts of DSBs with recruitment of 53BP1. Given that DNA repair is erroneous by nature, cooling-inflicted DNA damage may affect cell survival, proliferation, and genomic stability, significantly impacting cellular and organ function, with relevance in stem cell and transplantation procedures.https://doi.org/10.1177/09636897221108705 |
| spellingShingle | Marziyeh Tolouee Koen D. W. Hendriks Fia Fia Lie Lucas P. Gartzke Maaike Goris Femke Hoogstra-Berends Steven Bergink Robert H. Henning Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion Cell Transplantation |
| title | Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion |
| title_full | Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion |
| title_fullStr | Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion |
| title_full_unstemmed | Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion |
| title_short | Cooling of Cells and Organs Confers Extensive DNA Strand Breaks Through Oxidative Stress and ATP Depletion |
| title_sort | cooling of cells and organs confers extensive dna strand breaks through oxidative stress and atp depletion |
| url | https://doi.org/10.1177/09636897221108705 |
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