Ex Vivo Lung Perfusion of Rejected Human Donor Lungs; Are Donor Lungs with Prolonged Cold Ischemic Time Re-Conditioned by EVLP?

The application of ex vivo lung perfusion (EVLP) has significantly increased the successful clinical use of marginal donor lungs. While large animal EVLP models exist to test new strategies to improve organ repair, there is currently no rat EVLP model capable of maintaining long-term lung viability. Here, we describe a new rat EVLP model that addresses this need, while enabling the study of lung injury due to cold ischemic time (CIT). The technique involves perfusing and ventilating male Lewis rat donor lungs for 4 h before transplanting the left lung into a recipient rat and then evaluating lung function 2 h after reperfusion. To test injury within this model, lungs were divided into groups and exposed to different CITs (i.e., 20 min, 6 h, 12 h, 18 h and 24 h). Experiments involving the 24-h-CIT group were prematurely terminated due to the development of severe edema. For the other groups, no differences in the ratio of arterial oxygen partial pressure to fractional inspired oxygen ([Formula: see text]/[Formula: see text]) were observed during EVLP; however, lung compliance decreased over time in the 18-h group ( P = 0.012) and the [Formula: see text]/[Formula: see text] of the blood from the left pulmonary vein 2 h after transplantation was lower compared with 20-min-CIT group ( P = 0.0062). This new model maintained stable lung function during 4-h EVLP and after transplantation when exposed to up to 12 h of CIT.

Download Full-text

Transcriptomic investigation reveals donor specific gene signatures in human lung transplants

European Respiratory Journal ◽

10.1183/13993003.00327-2020 ◽

2020 ◽

pp. 2000327

Author(s):

Cristina Baciu ◽

Andrew Sage ◽

Ricardo Zamel ◽

Jason Shin ◽

Xiao-Hui Bai ◽

...

Keyword(s):

Gene Expression ◽

Ex Vivo ◽

Specific Gene ◽

Organ Shortage ◽

Ex Vivo Lung Perfusion ◽

Ischemic Time ◽

Protein Assay ◽

Cold Ischemic Time ◽

Donor Lung ◽

Lung Transplants

RationaleTransplantation of lungs from donation after circulatory death (DCD) in addition to donation after brain death (DBD) became routine worldwide to address the global organ shortage. The development of ex vivo lung perfusion (EVLP) for donor lung assessment and repair contributed to the increased use of DCD lungs. We hypothesise that better understanding of the differences between lungs from DBD and DCD donors, and between EVLP and directly transplanted (non-EVLP) lungs, will lead to discovery of the injury specific targets for donor lung repair and reconditioning.MethodsTissue biopsies from human DBD (n=177) and DCD (n=65) donor lungs assessed with or without EVLP, were collected at the end of cold ischemic time. All samples were processed with microarray assay. Gene expression, network and pathway analyses were performed using R, Ingenuity Pathway Analysis and STRING. Results were validated with protein assay, multiple logistic regression and 10-fold cross validation.ResultsOur analyses showed that lungs from DBD donors have up-regulation of inflammatory cytokines and pathways. In contrast, DCD lungs display a transcriptome signature of pathways associated with cell death, apoptosis and necrosis. Network centrality revealed specific drug targets to rehabilitate the DBD lungs. Moreover, in DBD lungs, TNFR1/2 signalling pathways and macrophage migration inhibitory factor associated pathways were activated in the EVLP group. A panel of genes that differentiate the EVLP from non-EVLP group in DBD lungs was identified.ConclusionThe examination of gene expression profiling indicates that DBD and DCD lungs have distinguishable biological transcriptome signatures.

Download Full-text

296 Time-Course Microarray Analysis of Rejected Human Donor Lungs during 12 Hours of Acellular Normothermic Ex Vivo Lung Perfusion

The Journal of Heart and Lung Transplantation ◽

10.1016/j.healun.2012.01.304 ◽

2012 ◽

Vol 31 (4) ◽

pp. S106-S107 ◽

Cited By ~ 2

Author(s):

J.C. Yeung ◽

P.C. Boutros ◽

R. Zamel ◽

M. Cypel ◽

X.-H. Bai ◽

...

Keyword(s):

Microarray Analysis ◽

Time Course ◽

Ex Vivo ◽

Lung Perfusion ◽

Human Donor ◽

Ex Vivo Lung Perfusion

Download Full-text

Subnormothermic ex vivo lung perfusion attenuates ischemia reperfusion injury from donation after circulatory death donors

PLoS ONE ◽

10.1371/journal.pone.0255155 ◽

2021 ◽

Vol 16 (8) ◽

pp. e0255155

Author(s):

Stephan Arni ◽

Tatsuo Maeyashiki ◽

Isabelle Opitz ◽

Ilhan Inci

Keyword(s):

Lung Tissue ◽

Ex Vivo ◽

Lung Perfusion ◽

Atp Content ◽

Donation After Circulatory Death ◽

Immunological Parameters ◽

Ex Vivo Lung Perfusion ◽

Ischemic Time ◽

Physiological Variables ◽

Circulatory Death

Use of normothermic ex vivo lung perfusion (EVLP) was adopted in clinical practice to assess the quality of marginal donor lungs. Subnormothermic perfusion temperatures are in use among other solid organs to improve biochemical, clinical and immunological parameters. In a rat EVLP model of donation after circulatory death (DCD) lung donors, we tested the effect of four subnormothermic EVLP temperatures that could further improve organ preservation. Warm ischemic time was of 2 hours. EVLP time was of 4 hours. Lung physiological data were recorded and metabolic parameters were assessed. Lung oxygenation at 21°C and 24°C were significantly improved whereas pulmonary vascular resistance and edema formation at 21°C EVLP were significantly worsened when compared to 37°C EVLP. The perfusate concentrations of potassium ions and lactate exiting the lungs with 28°C EVLP were significantly lower whereas sodium and chlorine ions with 32°C EVLP were significantly higher when compared to 37°C EVLP. Also compared to 37°C EVLP, the pro-inflammatory chemokines MIP2, MIP-1α, GRO-α, the cytokine IL-6 were significantly lower with 21°C, 24°C and 28°C EVLP, the IL-18 was significantly lower but only with 21°C EVLP and IL-1β was significantly lower at 21°C and 24°C EVLP. Compared to the 37°C EVLP, the lung tissue ATP content after 21°C, 24°C and 28°C EVLP were significantly higher, the carbonylated protein content after 28°C EVLP was significantly lower and we measured significantly higher myeloperoxidase activities in lung tissues with 21°C, 24°C and 32°C. The 28°C EVLP demonstrated acceptable physiological variables, significantly higher lung tissue ATP content and decreased tissue carbonylated proteins with reduced release of pro-inflammatory cytokines. In conclusion, the 28°C EVLP is a non inferior setting in comparison to the clinically approved 37°C EVLP and significantly improve biochemical, clinical and immunological parameters and may reduce I/R injuries of DCD lung donors.

Download Full-text

Better Oxygenation in Upper Lobes Than in Whole Lungs in Marginal Human Donor Lungs Utilized for Cellular Ex Vivo Lung Perfusion

The Journal of Heart and Lung Transplantation ◽

10.1016/j.healun.2019.01.808 ◽

2019 ◽

Vol 38 (4) ◽

pp. S321

Author(s):

H. Niikawa ◽

T. Okamoto ◽

K.S. Ayyat ◽

Y. Itoda ◽

C.F. Farver ◽

...

Keyword(s):

Ex Vivo ◽

Lung Perfusion ◽

Human Donor ◽

Ex Vivo Lung Perfusion

Download Full-text

Treatment of Cytomegalovirus in Human Donor Lungs with a Novel Chemokine-Based Immunotoxin during Ex Vivo Lung Perfusion Prevents Viral Reactivation

The Journal of Heart and Lung Transplantation ◽

10.1016/j.healun.2021.01.938 ◽

2021 ◽

Vol 40 (4) ◽

pp. S333

Author(s):

R. VP Ribeiro ◽

T. Ku ◽

V. H Ferreira ◽

L. Pires ◽

V. S Michaelsen ◽

...

Keyword(s):

Ex Vivo ◽

Lung Perfusion ◽

Viral Reactivation ◽

Human Donor ◽

Ex Vivo Lung Perfusion

Download Full-text

Porcine Model of Donation after Cardiac Death (DCD) and Evaluation of Pulmonary Function Using Ex-vivo Lung Perfusion (EVLP)

University of Ottawa Journal of Medicine ◽

10.18192/uojm.v9i1.4180 ◽

2019 ◽

Vol 9 (1) ◽

pp. 40-48

Author(s):

Malek Dhane

Keyword(s):

Weight Gain ◽

Cardiac Death ◽

Porcine Model ◽

Ex Vivo ◽

Lung Perfusion ◽

Warm Ischemia ◽

The Body ◽

Cold Ischemia ◽

Ex Vivo Lung Perfusion ◽

Ischemic Time

Objective The limiting factor when using lungs from donors after cardiac death (DCD) is the duration of warm ischemic time, which is linked to reperfusion edema. Within the context of lung shortages, and in order to avoid transplanting damaged lungs, ex-vivo lung perfusion (EVLP) has emerged as an innovative tool to preserve and recondition donor lungs. Using the EVLP technique in a porcine model, the purpose of this study is to determine the duration of warm ischemia that donor lungs can tolerate before they are viewed as non-viable for transplant. Methods This study is comprised of 5 groups (n=2-6/group). Four groups endured different warm ischemic times, whilst the fifth group underwent cold ischemia. The lungs were subsequently perfused outside the body using the EVLP platform for four hours. Results 120 minutes of warm ischemia is more damaging for lungs than 120 minutes of cold ischemia, even after being reconditioned on the EVLP platform (50,4 ± 8,9% vs. 3,3 ± 3,4% of weight gain). This would signify that two hours of warm ischemia is incompatible with transplantation. However, 90 minutes and 60 minutes of warm ischemia seems to have less of an effect on pulmonary functions. Indeed, the lungs of both these groups had less than 14% of weight gain and maintained oxygenating capacities (PaO2/FiO2 of 514 ± 12 and 586 ± 0 mmHg respectively.) Conclusion Lungs having been submitted to less than 90 minutes of warm ischemia and evaluated with EVLP may be suitable candidates for transplantation.

Download Full-text