Kinetic of gene expression signatures on ischemia porcine kidney
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ABSTRACT:
ORGANISM(S): Sus scrofa
PROVIDER: GSE109719 | GEO | 2018/02/10
REPOSITORIES: GEO
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Project description:Using an oligonucleotide microarray we performed differential transcriptomic analysis of porcine kidneys subjected to intense ischemic stress which could be observed in donors deceased after circulatory death situation (60 min warm ischemia then 24h of cold storage in University of Wisconsin solution) compared to healthy kidneys (n=3). 43 genes were differentially expressed in ischemic versus healthy kidneys (adjusted p value <0.05 + log2 fold change >0.5 or <-0.5). Functional enrichment analysis via Gene ontology revealed relevant biological processes and signaling pathways such as: cellular responses to stress and cell cycle adaptation, metabolism modification, RNA reprograming, cellular phenotype changes and inflammation. Our data showed that ischemia is a dynamic process, with important transcriptional modifications on major pathways. We uncovered a number of targets which we will further validate as biomarkers and therapeutic targets to optimize organ quality.
2016-03-22 | GSE79418 | GEO
Project description:Using an oligonucleotide microarray we performed differential transcriptomic analysis of porcine kidneys subjected to intense ischemic stress which could be observed in donors deceased after circulatory death situation (60 min warm ischemia then 24h of cold storage in University of Wisconsin solution) compared to healthy kidneys (n=3). 43 genes were differentially expressed in ischemic versus healthy kidneys (adjusted p value <0.05 + log2 fold change >0.5 or <-0.5). Functional enrichment analysis via Gene ontology revealed relevant biological processes and signaling pathways such as: cellular responses to stress and cell cycle adaptation, metabolism modification, RNA reprograming, cellular phenotype changes and inflammation. Our data showed that ischemia is a dynamic process, with important transcriptional modifications on major pathways. We uncovered a number of targets which we will further validate as biomarkers and therapeutic targets to optimize organ quality. ISCHEMIA samples: Three independent porcine kidney (from different Large White pig) were clamped, removed and maintained clamped 60 min at 37°C, and then flushed with cold (4°C) University of Wisconsin preservation solution (UW) and stored at 4°C for 24h, then kidneys were immediatly sampled and frozen. CONTROL samples: Three independent porcine kidney (from different Large White pig) were removed and then immediatly sampled and frozen.
2016-03-22 | E-GEOD-79418 | biostudies-arrayexpress
Project description:Despite a potentially huge number, uncontrolled donation after circulatory death contributed little to alleviating donor lung shortage due to rapidly progressive warm ischemia. Many methods have been studied in animals, but the tolerable warm ischemic time (WIT) remains less than 90 minutes. Using a refined mouse model of pulmonary artery ligation (PAL), we firstly determined the maximum tolerable WIT. 4-hour PAL caused mild lung infiltration without dysfunction upon reperfusion, whereas 5-hour PAL triggered arterial endothelium injury and more significant infiltration with dysfunction. Transcriptional profiling showed a myeloid-dominant inflammation with mild injury in 4-hour PAL. The maximum WIT was then adapted in a clinically relevant scenario. Donor mice died of circulatory arrest without heparization and remained at 37ºC for 4 hours, followed by isogenic transplantation. As observed in 4-hour PAL, nonhypoxic warm ischemia-reperfusion hardly affected graft function and histology, no matter if warm ischemic lung preserved gas exchange by spontaneously breathing or by postmortem protective ventilation. If the dead donors were left untouched, however, the grafted lungs suffered severe hypoxic warm ischemia-reperfusion injury, varying from partially aerated totally lost. Taken together, the retrieval time can be extended to 4 hours at 37ºC by preventing cardiac-dead donor lung hypoxia.
2022-03-30 | GSE154939 | GEO
Project description:We evaluated in a rat model of Donor after Circulatory Death (DCD) the effects of pretransplant kidney conditioning with Mesenchymal Stromal Cells (MSC) or MSC derived microvescicles (MV) on ischemic injury. Fisher rats (F) were used as kidney donors, Transgenic Sprague-Dawley rats expressing Enhanced Green Fluorescence Protein (EGFP) as MSC donors. After 20 min of warm ischemia nephrectomy was performed and kidneys were perfused with Belzer solution (BS), BS supplemented with MSC (MSC) or with MV (MV) for 4 h, at 4°C. Renal damage was evaluated by histology, renal gene expression by microarray analysis and RT-PCR. Malondialdehyde, lactate, LDH, glucose and pyruvate were measured in effluent fluid. MSC/MV kidneys showed a significant minor global ischemic damage and up-regulation of three genes encoding proteins that improve cell energy metabolism (Isocitrate dehydrogenase 2, NADH dehydrogenase Fe-S protein 8, Pyruvate dehydrogenase beta) and three genes encoding proteins involved in ion membrane transport (Calbindin 1, Monocarboxylate transporter 1, Vacuolar H+-ATPase d2 subunit). Lactate, LDH and malondialdehyde in effluent fluid were significantly lower in MSC/MV kidneys than BS. Glucose was lower while pyruvate higher in MSC/MV effluent than BS, suggesting a larger use of energy substrates by MSC/MV kidneys. MSC/MV perfusion of kidneys in addition to BS through HMP protects from ischemia injury by preserving the enzymatic machinery essential for cell viability and prepares kidney to reperfusion damage.
2017-07-21 | GSE84563 | GEO
Project description:In DCD organ donation one of the significant problems for the organ is the inflammatory response due to ischaemia reperfusion injury caused by warm ischaemia prior to retrieval followed by warm reoxygenation upon transplantation. We developed a model where a DCD kidney was retrieved and preserved using hypothermic machine perfusion with cold University of Wisconsin solution with/without the addition of heparinoids low molecular weight heparin and pentosan polysulfate (PPS) followed by warm oxygenated perfusion with modified Krebs-Henseleit buffer to simulate early ischaemia reperfusion injury (IRI). The donor rats consisted of a control group cold perfusion group compared to warm perfusion and groups treated with either low molecular weight heparin or PPS to assess their activity in ameliorating IRI. We used equimolar pooled RNA samples from each group to perform an exploratory microarray experiment Donor kidneys were harvested and preserved by hypothermic machine perfusion with University of Wisconsin (UW) Solution for 3 hours followed by warm oxygenated perfusion with modified Krebs-Henseleit buffer for 30 minutes to simulate transplantation and whole blood ischaemia reperfusion injury. The model was then used to test the anti-inflammatory properties of the glycosaminoglycan heparin and PPS when used as supplements into perfusate solutions. Half of the donor kidney was used to extract RNA using the Tri reagent and stored in RNA Later. The qualitative best three RNAs from each group were quantified by fluorimetry and mixed in an equimolar manner and used for microarray analysis.
2012-03-22 | E-GEOD-36671 | biostudies-arrayexpress
Project description:Deceased kidney donation after brain death (DBD) is the main source of transplants, yet these grafts yield inferior transplant outcomes when compared to living donation. In brain death, cerebral injury contributes to systemic biological dysregulation, causing significant cellular stress in donor kidneys that adversely impacts the quality of grafts. Here, we hypothesized that proteolytic processes in DBD kidneys might lead to podocyte damage with subsequent development of post-transplant dysfunction. Using protein topography and migration analysis platform (PROTOMAP), we mapped degradation profiles of cytoskeletal proteins in DBD kidneys. Cytoskeletal proteolytic degradation was further studied by Immunoblotting on a separate cohort of deceased and living donor kidney biopsies. To investigate potential mechanism of kidney cytoskeletal protein degradation, in-vitro human podocytes and ex-vivo precision-cut human kidney slices were employed. We found novel proteolytic profiles of key podocyte cytoskeletal proteins in donor kidneys associated with suboptimal posttransplant function. These were unique to brain-death and were not observed in circulatory-death or living-donor kidneys. Talin-specific protein degradation in DBD kidneys indicated Calpain-1 activation may have a key role in proteolytic processes observed in the dysfunctional kidneys. Investigation of the underlying mechanism suggests that Transforming-Growth Factor-β (TGFβ) induces Calpain-1 activation, leading to brain-death specific podocyte degradation patterns and dysregulation of actin cytoskeleton; events that were prevented, in-vitro, by Calpain inhibition. Conclusion Our data demonstrate that podocyte protein degradation impacts the quality of DBD kidneys, propose a role of TGFβ mediated Calpain-1 proteolytic processing of cytoskeletal Talin-1, suggesting therapeutic opportunities to prevent kidney dysfunction.
2022-08-12 | PXD022074 | Pride
Project description:Due to organ shortage, the transplantation community are increasingly using kidney from deceased donors such as donation after circulatory death (DCD) and donation after brain death (DBD). However, DCD donation have increased delayed graft function compared to DBD, an underline mechanism is still not well defied at the molecular level. In this study, we employed a rat model to mimic warm ischemia (45 mins) and reperfusion injury (IRI) for 4h and 24h. Apoptotic and tissue histological staining confirmed apoptosis and necrosis occurred at 4h and 24h post IRI respectively. Tissue proteome study revealed acute phase response, coagulation and complement activation and lipid X receptor activation as major pathway altered in intervention kidneys. Metabolomics follow up disclosed an increased level of lipids and fatty acids (FA). Mitochondrial function analysed by mitochondrial complex I activity and oxygen consumption and ATP levels in intervention kidney tissues were maintained 4h post IRI, but was significantly reduced 24h post IRI. Integrated proteo-metabolome analysis discovered an increased FA beta-oxidation 4h post IRI to sustain energy production. Kidney function were declined 24h post IRI indicated by increased blood creatinine and lactate levels. This study provides the frame work for the design of future metabolic intervention strategies to minimize kidney injury.
2020-07-06 | PXD005101 | Pride
Project description:Comparison of the gene expression in human MCA leading to the ischemic area from patients died due to ischemic stroke in comparison the the expression in the equivalent MCA in control subjects deceased due to extra cerebral causes.
2005-06-15 | E-MEXP-249 | biostudies-arrayexpress
Project description:Background: Strategies to improve long term renal allograft survival have been directed to recipient dependent mechanisms of renal allograft injury. In contrast, no such efforts have been made to optimize organ quality in the donor. In order to get insight into the deleterious gene pathways expressed at different time points during deceased kidney transplantation, transcriptomics was performed on kidney biopsies from a large cohort of deceased kidney transplants. Methods: A total of 554 kidney biopsies were taken from living and deceased donor kidneys at donation, after cold ischemia and after reperfusion. Transcriptomics by means of whole genome micro-array analyses followed by functional pathway analyses was performed. Results: Oxidative stress and complement- and coagulation pathways were uncovered as potential pathways for intervention in deceased donors. No genes were found to be differentially expressed between donation and cold ischemia. After reperfusion, pathways related to oxidative stress, NOD-like signalling, MAPK, cytokine-cytokine receptor, complement- and-coagulation and chemokines were enriched in kidneys from deceased organ donors. Pathways related to prolonged and worsening deprivation of oxygen were associated with delayed graft function of DCD grafts. Conclusions: The present study reveals oxidative stress and enrichment of complement and coagulation pathways in deceased donor kidneys. Future intervention therapies to optimize donor organ quality and prolong allograft survival should target oxidative stress and innate immune activation in the donor.
2015-04-02 | GSE43974 | GEO
Project description:In DCD organ donation one of the significant problems for the organ is the inflammatory response due to ischaemia reperfusion injury caused by warm ischaemia prior to retrieval followed by warm reoxygenation upon transplantation. We developed a model where a DCD kidney was retrieved and preserved using hypothermic machine perfusion with cold University of Wisconsin solution with/without the addition of heparinoids low molecular weight heparin and pentosan polysulfate (PPS) followed by warm oxygenated perfusion with modified Krebs-Henseleit buffer to simulate early ischaemia reperfusion injury (IRI). The donor rats consisted of a control group cold perfusion group compared to warm perfusion and groups treated with either low molecular weight heparin or PPS to assess their activity in ameliorating IRI. We used equimolar pooled RNA samples from each group to perform an exploratory microarray experiment
2012-03-22 | GSE36671 | GEO