Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes
Background. Normothermic ex vivo kidney perfusion (NEVKP) has demonstrated superior outcomes for donation-after-cardiovascular death grafts compared with static cold storage (SCS). To determine the mechanisms responsible for this, we performed an unbiased genome-wide microarray analysis. Methods. Ki...
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| Format: | Article |
| Language: | English |
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Wolters Kluwer
2021-08-01
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| Series: | Transplantation Direct |
| Online Access: | http://journals.lww.com/transplantationdirect/fulltext/10.1097/TXD.0000000000001157 |
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| author | Peter Urbanellis, MD, PhD, Caitriona M. McEvoy, MD, PhD Marko Škrtić, MD, PhD J. Moritz Kaths, MD, Dagmar Kollmann, MD, PhD Ivan Linares, MD, PhD Sujani Ganesh, MSc Fabiola Oquendo, MD Manraj Sharma, BSc (Hons) Laura Mazilescu, MD Toru Goto, MD Yuki Noguchi, MD, PhD Rohan John, MD Istvan Mucsi, MD, PhD Anand Ghanekar, MD, PhD Darius Bagli, MD Ana Konvalinka, MD, PhD Markus Selzner, MD Lisa A. Robinson, MD |
| author_facet | Peter Urbanellis, MD, PhD, Caitriona M. McEvoy, MD, PhD Marko Škrtić, MD, PhD J. Moritz Kaths, MD, Dagmar Kollmann, MD, PhD Ivan Linares, MD, PhD Sujani Ganesh, MSc Fabiola Oquendo, MD Manraj Sharma, BSc (Hons) Laura Mazilescu, MD Toru Goto, MD Yuki Noguchi, MD, PhD Rohan John, MD Istvan Mucsi, MD, PhD Anand Ghanekar, MD, PhD Darius Bagli, MD Ana Konvalinka, MD, PhD Markus Selzner, MD Lisa A. Robinson, MD |
| author_sort | Peter Urbanellis, MD, PhD, |
| collection | DOAJ |
| description | Background. Normothermic ex vivo kidney perfusion (NEVKP) has demonstrated superior outcomes for donation-after-cardiovascular death grafts compared with static cold storage (SCS). To determine the mechanisms responsible for this, we performed an unbiased genome-wide microarray analysis.
Methods. Kidneys from 30-kg Yorkshire pigs were subjected to 30 min of warm ischemia followed by 8 h of NEVKP or SCS, or no storage, before autotransplantation. mRNA expression was analyzed on renal biopsies on postoperative day 3. Gene set enrichment analysis was performed using hallmark gene sets, Gene Ontology, and pathway analysis.
Results. The gene expression profile of NEVKP-stored grafts closely resembled no storage kidneys. Gene set enrichment analysis demonstrated enrichment of fatty acid metabolism and oxidative phosphorylation following NEVKP, whereas SCS-enriched gene sets were related to mitosis, cell cycle checkpoint, and reactive oxygen species (q < 0.05). Pathway analysis demonstrated enrichment of lipid oxidation/metabolism, the Krebs cycle, and pyruvate metabolism in NEVKP compared with SCS (q < 0.05). Comparison of our findings with external data sets of renal ischemia-reperfusion injury revealed that SCS-stored grafts demonstrated similar gene expression profiles to ischemia-reperfusion injury, whereas the profile of NEVKP-stored grafts resembled recovered kidneys.
Conclusions. Increased transcripts of key mitochondrial metabolic pathways following NEVKP storage may account for improved donation-after-cardiovascular death graft function, compared with SCS, which promoted expression of genes typically perturbed during IRI. |
| first_indexed | 2024-12-17T01:16:46Z |
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| institution | Directory Open Access Journal |
| issn | 2373-8731 |
| language | English |
| last_indexed | 2024-12-17T01:16:46Z |
| publishDate | 2021-08-01 |
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| series | Transplantation Direct |
| spelling | doaj.art-d110c16741be4afcab2dde2ba45f09812022-12-21T22:08:59ZengWolters KluwerTransplantation Direct2373-87312021-08-0178e71910.1097/TXD.0000000000001157202108000-00001Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism GenesPeter Urbanellis, MD, PhD,0Caitriona M. McEvoy, MD, PhD1Marko Škrtić, MD, PhD2J. Moritz Kaths, MD,3Dagmar Kollmann, MD, PhD4Ivan Linares, MD, PhD5Sujani Ganesh, MSc6Fabiola Oquendo, MD7Manraj Sharma, BSc (Hons)8Laura Mazilescu, MD9Toru Goto, MD10Yuki Noguchi, MD, PhD11Rohan John, MD12Istvan Mucsi, MD, PhD13Anand Ghanekar, MD, PhD14Darius Bagli, MD15Ana Konvalinka, MD, PhD16Markus Selzner, MD17Lisa A. Robinson, MD181 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.5 Division of Nephrology, Department of Medicine, University of Toronto, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.7 Laboratory Medicine and Pathobiology, Toronto General Hospital, University of Toronto, Toronto, ON, Canada.2 Canadian Donation and Transplantation Research Program, Edmonton, AB, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.8 Departments of Surgery (Urology) and Physiology, The Hospital for Sick Children, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.1 Soham and Shaila Ajmera Family Transplant Centre, Toronto General Hospital, University Health Network, Toronto, ON, Canada.6 Program in Cell Biology, The Hospital for Sick Children Research Institute, Toronto, ON, Canada.Background. Normothermic ex vivo kidney perfusion (NEVKP) has demonstrated superior outcomes for donation-after-cardiovascular death grafts compared with static cold storage (SCS). To determine the mechanisms responsible for this, we performed an unbiased genome-wide microarray analysis. Methods. Kidneys from 30-kg Yorkshire pigs were subjected to 30 min of warm ischemia followed by 8 h of NEVKP or SCS, or no storage, before autotransplantation. mRNA expression was analyzed on renal biopsies on postoperative day 3. Gene set enrichment analysis was performed using hallmark gene sets, Gene Ontology, and pathway analysis. Results. The gene expression profile of NEVKP-stored grafts closely resembled no storage kidneys. Gene set enrichment analysis demonstrated enrichment of fatty acid metabolism and oxidative phosphorylation following NEVKP, whereas SCS-enriched gene sets were related to mitosis, cell cycle checkpoint, and reactive oxygen species (q < 0.05). Pathway analysis demonstrated enrichment of lipid oxidation/metabolism, the Krebs cycle, and pyruvate metabolism in NEVKP compared with SCS (q < 0.05). Comparison of our findings with external data sets of renal ischemia-reperfusion injury revealed that SCS-stored grafts demonstrated similar gene expression profiles to ischemia-reperfusion injury, whereas the profile of NEVKP-stored grafts resembled recovered kidneys. Conclusions. Increased transcripts of key mitochondrial metabolic pathways following NEVKP storage may account for improved donation-after-cardiovascular death graft function, compared with SCS, which promoted expression of genes typically perturbed during IRI.http://journals.lww.com/transplantationdirect/fulltext/10.1097/TXD.0000000000001157 |
| spellingShingle | Peter Urbanellis, MD, PhD, Caitriona M. McEvoy, MD, PhD Marko Škrtić, MD, PhD J. Moritz Kaths, MD, Dagmar Kollmann, MD, PhD Ivan Linares, MD, PhD Sujani Ganesh, MSc Fabiola Oquendo, MD Manraj Sharma, BSc (Hons) Laura Mazilescu, MD Toru Goto, MD Yuki Noguchi, MD, PhD Rohan John, MD Istvan Mucsi, MD, PhD Anand Ghanekar, MD, PhD Darius Bagli, MD Ana Konvalinka, MD, PhD Markus Selzner, MD Lisa A. Robinson, MD Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes Transplantation Direct |
| title | Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes |
| title_full | Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes |
| title_fullStr | Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes |
| title_full_unstemmed | Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes |
| title_short | Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex Vivo Perfusion Demonstrates an Enrichment of Mitochondrial Metabolism Genes |
| title_sort | transcriptome analysis of kidney grafts subjected to normothermic ex vivo perfusion demonstrates an enrichment of mitochondrial metabolism genes |
| url | http://journals.lww.com/transplantationdirect/fulltext/10.1097/TXD.0000000000001157 |
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