Human ex vivo Lung Perfusion: A Pre-Clinical Model for Lung Disease
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE69416 | GEO | 2015/06/02
SECONDARY ACCESSION(S): PRJNA285460
REPOSITORIES: GEO
Similar Datasets
Project description:Ex vivo lung perfusion (EVLP) has extended the number of transplantable lungs by reconditioning marginal organs. However EVLP is performed at 37°C without homeostatic regulation leading to metabolic wastes accumulation in the perfusate, and as a corrective measure, the costly perfusate is repeatedly replaced during the standard of care procedure. As an interesting alternative, a hemodialyzer could be placed on the EVLP circuit, that was previously shown to rebalance the perfusate composition, maintain lung function and viability without appearing to impact the global gene expression in the lung. Here, we assessed the biological effects of a hemodialyzer during EVLP by performing biochemical and refined functional genomic analyses over a 12 h procedure in a pig model. We found that dialysis stabilized electrolytic and metabolic parameters of the perfusate but enhanced the gene expression and protein accumulation of several inflammatory cytokines, and promoted a genomic profile predicting higher endothelial activation already at 6 h and immune cytokine signaling at 12 h. Therefore epuration of EVLP with a dialyzer, while correcting features of perfusate composition and maintaining respiratory function, promotes inflammatory responses in the tissue. This finding suggests that modifying the metabolite composition of the perfusate by dialysis during EVLP can have detrimental effects on the tissue response and that this strategy should not be transferred as such to the clinic.
2024-03-04 | GSE241921 | GEO
Project description:In the present study, we demonstrate the feasibility of the EVLP technique to perfuse lungs from deceased patients with a broad spectrum of chronic lung diseases. We were able to assess gene expression changes over the course of EVLP and between end-stage diseases by RNA sequencing transcriptomics from tissue samples collected at different time points, revealing fundamental genetic information which has important avenues for modulation in the future. Thus, EVLP can serve as a clinically relevant experimental model to derive mechanistic insights into pulmonary pathophysiology and various human lung diseases.
2021-01-21 | GSE146436 | GEO
Project description:Ischemia reperfusion (IR) injury is a key complication following lung transplantation, and it remains to be fully resolved. Ex vivo lung perfusion (EVLP) has shown its significance as a platform to assess and repair marginal donor lungs prior to transplants with good clinical outcomes. Recently, it has been shown that Glutamax supplementation to Steen solution can improve basic cellular function as well as porcine EVLP duration with stable lung quality. To better understand the molecular mechanisms of IR injury, acellular EVLP, and Glutamax supplementation during EVLP, transcriptomic changes in cell culture models simulating IR and EVLP have been investigated. Briefly, human pulmonary microvascular endothelial cells (HPMEC) and human lung epithelial cells (BEAS-2B) were cultured in DMEM+10%FBS, then were perfused with Perfadex at 4°C for 18H to simulate cold ischemic time (CIT). Subsequently, the cells were perfused in either DMEM+10%FBS (IR model) or in Steen solution or Steen with 4mM Glutamax (EVLP models) for 4H. RNA samples were collected for CIT, IR, and two EVLP conditions, and sequencing data were analyzed for differentially expressed genes and pathways. HPMEC and BEAS-2B cells showed significant changes in gene expressions for IR and EVLP models. In pathway analyses, IR models of both cell types had up-regulated pro-inflammatory responses and down-regulated cell metabolism, and there was up-regulation of epithelial process and/or vascular activation. Steen alone EVLP models exhibited down-regulation of cell metabolism, with an absence of inflammatory and vascular or epithelial signals, while Steen with Glutamax exhibited similar down-regulation of cell metabolism as well as down-regulated inflammatory and vascular process. The commonly used EVLP perfusate, acellular Steen solution, can silence the activation of pro-inflammatory signaling in HPMEC and BEAS-2B cell culture models. This finding may yield insights into how acellular EVLP has been successful in safe preservation of the donor lungs during organ evaluation and repair prior to lung transplantation.
2023-05-30 | GSE228488 | GEO
Project description:The field of graft preservation has made considerable strides in recent years improving outcomes related to solid organ restoration and regeneration. In lungs, the use of ex vivo lung perfusion (EVLP) in line with devices and treatments has shown promising results within preclinical and clinical studies with the potential to improve graft quality. The benefit of the therapy would be to render marginal and declined donor lungs suitable for transplantation, ultimately increasing the donor pool available for transplantation. Additionally, such therapies used in machine perfusion could also increase preservation time, facilitating logistical planning. Cytokine adsorption has been demonstrated as a potentially safe and effective therapy when applied to the EVLP circuit and post transplantation. The mechanism by which this treatment improves the donor lung on a molecular basis is not yet fully elucidated. We hypothesized that there were characteristic inflammatory and immunomodulatory differences between lungs treated with and without cytokine adsorption, reflecting in proteomic changes in gene ontology pathways and across inflammation-related proteins. In the current study we investigate the molecular mechanisms and signaling pathways of how cytokine adsorption impacts the lung function when used during EVLP and when used post transplantation as hemoperfusion in a porcine model. Lung tissue from EVLP and post lung transplantation were analyzed for their proteomic profile using mass spectrometry. The inflammatory and immune processes were compared between the treated and the non-treated groups to show the differences occurring between the forms of graft preservation.
2023-10-24 | PXD044413 | Pride
Project description:Proteins that were newly-synthesized during ex vivo lung perfusion were labeled with an azido-sugar Ac4GalNAz in the perfusate. These proteins were enriched using click chemistry to attach an alkyne-desthiobiotin group to the azido-labeled proteins, then pulled out of solution using streptavidin beads. The overall goal is to better understand the effects of warm ischemia injury in lungs destined for lung transplantation.
2022-10-07 | MSV000090491 | MassIVE
Project description:Ex vivo lung perfusion restores normothermia, ventilation and circulation to donor lungs, typically after a period of cold ischemia. This allows donor lungs to be evaluated prior to transplantation. We used microarrays to study the biological response of human lungs to Ex Vivo Lung Perfusion. Samples were collected from donor lungs at Toronto General Hospital. Lungs were donation after brain death (DBD)
2020-06-22 | GSE127055 | GEO
Project description:Ex vivo lung perfusion (EVLP) is a largely employed technique in transplantation that permits the reassessment of marginal lungs, thereby increasing the graft pool. However its biological effects on the different lung cell types are not known. In order to advance our mechanistic understanding and further improve EVLP, we conducted a pioneer study involving single-cell RNA-seq (scRNA-seq) on two human lungs declined for transplantation. We integrated data sets generated at 4 h (clinical duration) and 10 h (prolonged duration) and revealed functional changes common to both lungs that include inflammation in most cell types, loss of barrier function in epithelial and endothelial cells especially at 10 h, and reduction of migration properties in immune cells. In addition to providing insights into the biological effects of EVLP, scRNA-seq applied to EVLP offers a highly powerful way to evaluate novel EVLP therapeutic advances that will benefit to lung transplantation.
2023-06-20 | GSE218788 | GEO
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
Project description:There is a ever widening gap between the availability of donor livers suitable for transplantation and liver transplant demand of a growing waiting list. To expand the pool of liver grafts, donation after circulatory death is increasingly being used. Livers procured after circulatory death are exposed to a harmful period of warm Ischemia (WI) during the donation process, reducing post-transplant results significantly. Additionally, the conventional technique of static cold storage does not protect grafts that suffered WI. New dynamic preservation strategies by means of Normothermic Machine Perfusion (NMP) showed enhanced preservation of these type of grafts, but it is unknown how NMP influences liver cell biology during perfusion. Hepatocytes and cholnagiocytes have been shown to release Extracellular Vesicles (EVs) in the systemic circulation and bile, respectively. We hypothesized that EVs are released during liver NMP as well, in perfusate and bile. EVs have been separated from perfusate and bile during NMP of porcine livers, and the RNA cargo of the EVs has been sequenced to gain insights on liver cell biology during machine perfusion. Pigs were randomly allocated to undergo liver procurement either immediately after the start of surgery (no-WI, n=5) or after a period of WI of 60-minutes (clamping of thoracic aorta; WI-60, n=5). All livers (n=10) were cooled down and flushed out with ice-cold preservation solution, according to standard clinical practice. Porcine blood was collected and washed with a cell saver to obtain packed red blood cells for NMP. The hepatic artery, portal vein, and inferior vena cava were cannulated and after a period of about 2-hours of cold storage, NMP was started. All liver underwent NMP for 6-hours. Packed red blood-based perfusate samples were taken at 1, 3, and 6h of NMP; whereas, the bile produced the first 3-hours and the last 3-hours was pooled in 2 samples. Perfusate and bile samples were processed immediately for EVs separation with charge-based precipitation in a mixture of protamine and polyethylene glycol. The total RNA was extracted from EVs. Libraries of mRNA and of small RNA were prepared and analyzed for quality and size range, and sequenced on an Illumina HiSeq4000 instrument. Count-based differential expression analysis was done with R-based Bioconductor package DESeq2, and the following comparisons were performed: - Unpaired (between groups) comparison - perfusate samples WI-60 vs. no-WI - bile samples WI-60 vs. no-WI - Paired (withing group) comparison - no-WI - perfusate sample 1h vs. perfusate sample 3h - perfusate sample 1-3h vs. perfusate sample 6h - bile sample 1-3h vs. bile sample 3-6h - perfusate samples vs. bile samples - WI-60 - bile sample 1-3h vs. bile sample 3-6h - perfusate samples vs. bile samples Results were adjusted for multiple testing with the Benjamini-Hochberg procedure, to control for false discovery rate. Additionally, the function of the transcripts identified in the EVs of both groups and of the differentially expressed transcripts was investigated with gene ontology enrichment analysis for the domains \\"biological process\\" and \\"pathways\\".
2024-09-27 | E-MTAB-11420 | biostudies-arrayexpress
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