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Transcriptomic analysis of Kidney Machine normothermic perfusion and comparision to cold storage

ABSTRACT: Kidneys which had been taken for transplant but later declined were perfused for up to 4 hours using a normothermic perfusion (MNP) rig or left on ice (CS) and samples were taken for RNASeq at different time points. The effect of the addition of a heamoadsorber (HA) to the perfusion rig was also assessed.

ORGANISM(S): Homo sapiens

PROVIDER: GSE121447 | GEO | 2020/10/17

REPOSITORIES: GEO

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Mechanisms and treatment of cold-storage-induced microvascular obstructions in normothermic machine perfusion of human kidney

Project description:Human kidneys which were accepted for transplantation and subsequently not used were subjected to ex vivo normothermic machine perfusion. During perfusion it was found the tubular epithelia synthesized fibrinogen which led to red blood cell aggregation and pathologic plugging of renal microvasculature. Sequential ex vivo delivery of plasminogen and tissue plasminogen activator (tPA) during normothermic machine perfusion effectively lyses these fibrinogen-mediated plugs.

2022-10-01 | GSE139017 | GEO

Transcriptomic analysis of Kidney Machine normothermic perfusion and comparision to cold storage

Project description:Transcriptomic analysis of Kidney Machine normothermic perfusion and comparision to cold storage

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Differential expressed genes in biopsies from discarded human kidneys after one hour of normothermic perfusion

Project description:Nanostring nCounter Organ Transplant Panel data on 4 cortex biopsies from discarded human kidneys analysing the differential expressed genes after one hour of normothermic perfusion

2022-02-24 | GSE197304 | GEO

Pan-caspase inhibition during normothermic machine perfusion of discarded livers mitigates ischemia-reperfusion injury

Project description:Access to liver transplantation is limited by a significant organ shortage. The recent introduction of machine perfusion technology allows surgeons to monitor and assess ex situ liver function prior to transplantation. However, many donated organs are of inadequate quality for transplant, though opportunities exist to rehabilitate organ function with adjunct therapeutics during normothermic machine perfusion. In this preclinical study, we targeted the apoptosis pathway as a potential method of improving hepatocellular function. Treatment of discarded human livers during normothermic perfusion with an irreversible pan-caspase inhibitor, emricasan, resulted in significant mitigation of ischemia-reperfusion injury at both the transcriptional and protein level. This was evidenced by significantly decreased circulating levels of the pro-inflammatory cytokines, interleukin-6, interleukin-8, and interferon-gamma, compared to control livers. Untreated livers also demonstrated transcriptional changes notable for enrichment in pathways involved in innate immunity, leukocyte migration, and cytokine-mediated signaling. Targeting of unregulated apoptosis may represent a viable therapeutic intervention for rehabilitating liver hepatocellular function during machine perfusion.

2022-07-11 | GSE202565 | GEO

Transcriptome Analysis of Kidney Grafts Subjected to Normothermic Ex-Vivo Kidney Perfusion

Project description:Normothermic ex-vivo kidney perfusion (NEVKP) has demonstrated superior outcomes for donation-after-cardiovascular death (DCD) grafts compared to static cold storage (SCS). To determine the mechanisms responsible for this, we performed an unbiased genome-wide microarray analysis. Kidneys from 30kg-Yorkshire pigs were subjected to 30min of warm ischemia followed by 8hrs of NEVKP or SCS, or no storage (NS), prior to auto-transplantation. mRNA expression was analyzed on POD3 renal biopsies.

2021-07-15 | GSE155418 | GEO

Protein Expression in Liver Tissue and Bile of Elderly Sprague-Dawley Rats after Normothermic Machine Perfusion

Project description:Liver are frequently declined for transplantation due to the donor age. It is still unclear how donor age affects the graft quality. Normothermic machine perfusion (NMP) has been proposed as a useful assessment tool prior to transplantation. We aimed to compare the performance of young and elderly rat liver grafts in a small animal NMP model. Grafts from 24 rats were procured, either 3 or 12 months old and perfused for 6 hours at 37°C or stored on ice as a reference group (n=6/group). Livers in both NMP groups cleared lactate and produced bile, with similar pressure, bile production, and pH levels. However, older rat livers showed higher peak transaminase and urea levels. Proteomic analysis revealed differences in protein composition, particularly higher levels of proteins related to oxidoreductase and catalytic activity in older livers. Older age appeared to increase susceptibility to oxidative stress in the livers.

2024-08-09 | PXD045394 | Pride

Gene expression profiling of human livers during normothermic machine perfusion

Project description:Normothermic machine perfusion (NMP) has been successfully implemented in clinical routine of liver transplantation over the past years. However, little is known about the mechanisms how NMP impacts on the transcriptome of a human donor liver. We herein examined gene expression profiles in transplanted and non-transplanted livers over NMP time. 50 livers subjected to NMP were included in this study. 30 were transplanted after a maximum of 20 hours (h) perfusion, while 15 were discarded due to poor performance. Biopsies were collected befor eNP (PRE), 1h, 6h, 12h, 20h of NMP and after reperfusion. Next-generation sequencing was applied in liver biopsies to assess differential gene expression over perfusion time. Perfusate samples were collected regularly to monitor liver function. Comparison in differential gene expression between PRE and 20h NMP showed 415 upregulated and 727 downregulated genes. Most significantly upregulated genes were associated with extra cellular matrix organization, cell growth/differentiation processes and cytokine signaling. A set of genes were identified which were significantly differentially expressed and important for classification of non-transplanted vs transplanted biopsies, especially at 12 and 20h of NMP. A 7-gene-signature showed good separation already at 6H NMP, thereby CD274 (PD-L1) expression was ponted out as most important.

2024-08-29 | GSE263614 | GEO

Gene expression profiling of human livers during normothermic machine perfusion

Project description:Gene expression profiling of human livers during normothermic machine perfusion

| PRJNA1098409 | ENA

Urine recirculation prolongs normothermic kidney perfusion for via more optimal metabolic homeostasis – a proteomics study

Project description:We describe a proteomics analysis to determine the molecular differences between normothermically perfused (normothermic machine perfusion, NMP) human kidneys with urine recirculation (URC) and urine replacement (UR). Proteins were extracted from 16 kidney biopsies with URC (n=8 donors after brain death (DBD), n=8 donors after circulatory death (DCD)) and three with UR (n=2 DBD, n=1 DCD), followed by quantitative analysis by mass spectrometry. Damage-associated molecular patterns (DAMPs) were decreased in kidney tissue after six hours NMP with URC, suggesting reduced inflammation. Vasoconstriction was also attenuated in kidneys with URC as angiotensinogen levels were reduced. Strikingly, kidneys became metabolically active during NMP, which could be enhanced and prolonged by URC. For instance, mitochondrial succinate dehydrogenase enzyme levels as well as carbonic anhydrase were enhanced with URC, contributing to pH stabilisation. Levels of cytosolic and the mitochondrial phosphoenolpyruvate carboxykinase were elevated after 24 hours of NMP, more prevalent in DCD than DBD tissue. Key enzymes involved in glucose metabolism were also increased after twelve and 24 hours of NMP with URC, including mitochondrial malate dehydrogenase and glutamic-oxaloacetic transaminase, predominantly in DCD tissue. We conclude that NMP with URC permits prolonged preservation and revitalises metabolism to possibly minimize better cope with ischemia reperfusion injury in discarded kidneys.

2020-10-20 | PXD021507 | Pride

Inflammatory gene expression in livers undergoing ex situ normothermic perfusion is attenuated by leukocyte removal from the perfusate

Project description:ABSTRACT: Background. Ex situ normothermic perfusion (ESNP) is a method to evaluate and potentially recondition organs before transplantation. However, increased expression of inflammatory molecules, including by tissue-resident immune cells, may occur during the perfusion process, potentially negating the beneficial effects of perfusion. Methods. We used RNA sequencing to assess gene expression in 31 livers undergoing ESNP, including 23 donated after circulatory death (DCD) and 8 donated after brain death. In 7 DCD livers, a leucocyte filter was added to the circuit during perfusion. Biopsies were available for transcriptomic assessment in all cases at the start of perfusion and at varying time points postperfusion. Results. During ESNP in DCD livers, we observed an increase in proinflammatory, profibrinolytic, and prorepair pathway genes. SERPINE1, encoding plasminogen activator inhibitor-1, was among the genes most significantly upregulated during perfusion in DCD livers, potentially promoting fibrin clot persistence in vasculature. We also found increased expression of monocyte and neutrophil recruiting chemokine and proinflammatory cytokine transcripts during ESNP, but several prorepair molecules, including thymic stromal lymphopoietin, were also upregulated. In both DCD and donation after brain death livers, interferon-gamma response genes were enriched, whereas oxidative phosphorylation genes decreased in organs with high perfusate alanine transaminase, a biomarker associated with adverse clinical outcomes. The inclusion of a leukocyte filter in the perfusion circuit mitigated the induction of inflammation/immune pathway genes during perfusion and was associated with enrichment in oxidative phosphorylation genes. Conclusions. Leukocyte removal during ESNP abrogates transcriptional changes that are associated with unfavorable clinical outcomes, potentially benefiting human livers undergoing ESNP.

2024-10-01 | GSE245976 | GEO

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