The Effects of Free Heme on Functional and Molecular Changes During Normothermic Machine Perfusion of Human Kidneys

Overview

Journal Front Immunol

Specialty Allergy & Immunology

Date 2022 May 19

PMID 35585981

Authors

Sarah A Hosgood

Tegwen R Elliott

Nina P Jordan

Michael L Nicholson

Affiliations

Soon will be listed here.

Abstract

Normothermic machine perfusion (NMP) is a technique of kidney preservation designed to restore cellular metabolism after cold ischemia. Kidneys are perfused with an oxygenated banked red blood cell (RBC) based solution for 1h at 36°C. During NMP, RBCs can become damaged, releasing free heme into the perfusate. This can act as a damage-associated molecular pattern (DAMP) activating inflammatory signalling pathways. The aim of this study was to measure the levels of free heme during NMP, assess the effect on kidney function and determine any association with inflammatory and stress related gene expression. Levels of free heme were measured in perfusate samples from a series of donation after circulatory death (DCD) kidneys undergoing NMP as part of a randomised controlled trial (RCT). The age of RBCs and levels of free heme were correlated with perfusion parameters. Changes in gene expression were analysed in a series of kidneys declined for transplantation using the NanoString nCounter Organ Transplant Panel and qRT-PCR. Older units of RBCs were associated with higher levels of free heme and levels increased significantly during NMP (Pre 8.56 ± 7.19µM vs 26.29 ± 15.18µM, P<0.0001). There was no association with levels of free heme and perfusion parameters during NMP (P > 0.05). Transcriptional and qPCR analysis demonstrated the upregulation of differentially expressed genes associated with apoptosis (FOS and JUN), inflammatory cytokines (IL-6, SOCS3, ATF3), chemokines (CXCL8, CXCL2, CC3/L1) and oxidative stress (KLF4) after NMP. However, these did not correlate with levels of free heme (P >0.05). A significant amount of free heme can be detected in the perfusate before and after NMP particularly when older units of red cells are used. Although transcriptional analysis demonstrated significant upregulation of genes involved with apoptotic, inflammatory and oxidative pathways these were not associated with high levels of free heme.

Citing Articles

Oxygen carriers affect kidney immunogenicity during machine perfusion.

Rother T, Horgby C, Schmalkuche K, Burgmann J, Nocke F, Jagers J Front Transplant. 2024; 2:1183908.

PMID: 38993849 PMC: 11235266. DOI: 10.3389/frtra.2023.1183908.

normothermic preservation of a kidney graft from uncontrolled donation after circulatory death over 73 hours.

Montagud-Marrahi E, Luque Y, Ros R, Ajami T, Cuadrado-Payan E, Estrella H Front Bioeng Biotechnol. 2024; 11:1330043.

PMID: 38283171 PMC: 10811075. DOI: 10.3389/fbioe.2023.1330043.

Pushing the boundaries of innovation: the potential of organ perfusion from an interdisciplinary point of view.

Iske J, Schroeter A, Knoedler S, Nazari-Shafti T, Wert L, Roesel M Front Cardiovasc Med. 2023; 10:1272945.

PMID: 37900569 PMC: 10602690. DOI: 10.3389/fcvm.2023.1272945.

Current Evidence and Future Perspectives to Implement Continuous and End-Ischemic Use of Normothermic and Oxygenated Hypothermic Machine Perfusion in Clinical Practice.

Foguenne M, MacMillan S, Kron P, Nath J, Devresse A, De Meyer M J Clin Med. 2023; 12(9).

PMID: 37176647 PMC: 10178893. DOI: 10.3390/jcm12093207.

How to Best Protect Kidneys for Transplantation-Mechanistic Target.

Akalay S, Hosgood S J Clin Med. 2023; 12(5).

PMID: 36902572 PMC: 10003664. DOI: 10.3390/jcm12051787.

References

Hosgood S, Thompson E, Moore T, Wilson C, Nicholson M . Normothermic machine perfusion for the assessment and transplantation of declined human kidneys from donation after circulatory death donors. Br J Surg. 2017; 105(4):388-394. PMC: 5887977. DOI: 10.1002/bjs.10733. View

Himbert S, Qadri S, Sheffield W, Schubert P, DAlessandro A, Rheinstadter M . Blood bank storage of red blood cells increases RBC cytoplasmic membrane order and bending rigidity. PLoS One. 2021; 16(11):e0259267. PMC: 8589153. DOI: 10.1371/journal.pone.0259267. View

Arni S, Necati C, Maeyashiki T, Opitz I, Inci I . Perfluorocarbon-Based Oxygen Carriers and Subnormothermic Lung Machine Perfusion Decrease Production of Pro-Inflammatory Mediators. Cells. 2021; 10(9). PMC: 8466246. DOI: 10.3390/cells10092249. View

Florim G, Caldas H, Goncalves N, Bueno G, Baptista M, Fernandes-Charpiot I . Activation of HMGB1-TLR4 Pathway and Inflammasome Contribute to Enhanced Inflammatory Response in Extended Criteria and Kidneys With KDPI ≥85. Transplantation. 2019; 104(4):724-730. DOI: 10.1097/TP.0000000000003048. View

Vallelian F, Schaer C, Deuel J, Ingoglia G, Humar R, Buehler P . Revisiting the putative role of heme as a trigger of inflammation. Pharmacol Res Perspect. 2018; 6(2):e00392. PMC: 5878102. DOI: 10.1002/prp2.392. View

Vercaemst L . Hemolysis in cardiac surgery patients undergoing cardiopulmonary bypass: a review in search of a treatment algorithm. J Extra Corpor Technol. 2009; 40(4):257-67. PMC: 4680715. View

Zlatev H, von Horn C, Kaths M, Paul A, Minor T . Clinical use of controlled oxygenated rewarming of kidney grafts prior to transplantation by ex vivo machine perfusion. A pilot study. Eur J Clin Invest. 2021; 52(2):e13691. DOI: 10.1111/eci.13691. View

Zhao Z, Hu Z, Zeng R, Yao Y . HMGB1 in kidney diseases. Life Sci. 2020; 259:118203. DOI: 10.1016/j.lfs.2020.118203. View

Garcia-Roa M, Vicente-Ayuso M, Bobes A, Pedraza A, Gonzalez-Fernandez A, Martin M . Red blood cell storage time and transfusion: current practice, concerns and future perspectives. Blood Transfus. 2017; 15(3):222-231. PMC: 5448828. DOI: 10.2450/2017.0345-16. View

10.

Urbanellis P, Hamar M, Kaths J, Kollmann D, Linares I, Mazilescu L . Normothermic Ex Vivo Kidney Perfusion Improves Early DCD Graft Function Compared With Hypothermic Machine Perfusion and Static Cold Storage. Transplantation. 2019; 104(5):947-955. DOI: 10.1097/TP.0000000000003066. View

11.

Zakkar M, Guida G, Suleiman M, Angelini G . Cardiopulmonary bypass and oxidative stress. Oxid Med Cell Longev. 2015; 2015:189863. PMC: 4334937. DOI: 10.1155/2015/189863. View

12.

Bhattacharjee R, Patel S, Sun Q, Jiang L, Richard-Mohamed M, Ruthirakanthan A . Renal Protection Against Ischemia Reperfusion Injury: Hemoglobin-based Oxygen Carrier-201 Versus Blood as an Oxygen Carrier in Ex Vivo Subnormothermic Machine Perfusion. Transplantation. 2019; 104(3):482-489. DOI: 10.1097/TP.0000000000002967. View

13.

Helms C, Gladwin M, Kim-Shapiro D . Erythrocytes and Vascular Function: Oxygen and Nitric Oxide. Front Physiol. 2018; 9:125. PMC: 5826969. DOI: 10.3389/fphys.2018.00125. View

14.

Habib R . Multifaceted roles of Toll-like receptors in acute kidney injury. Heliyon. 2021; 7(3):e06441. PMC: 7944035. DOI: 10.1016/j.heliyon.2021.e06441. View

15.

Chiabrando D, Mercurio S, Tolosano E . Heme and erythropoieis: more than a structural role. Haematologica. 2014; 99(6):973-83. PMC: 4040894. DOI: 10.3324/haematol.2013.091991. View

16.

Hao J, Li Y, Li R . A novel mechanism of NALP3 inducing ischemia reperfusion injury by activating MAPK pathway in acute renal failure. Med Hypotheses. 2013; 80(4):463-5. DOI: 10.1016/j.mehy.2012.12.041. View

17.

Cardigan R, New H, Tinegate H, Thomas S . Washed red cells: theory and practice. Vox Sang. 2020; 115(8):606-616. DOI: 10.1111/vox.12971. View

18.

Ichikawa J, Koshino I, Arashiki N, Nakamura F, Komori M . Storage-Related Changes in Autologous Whole Blood and Irradiated Allogeneic Red Blood Cells and Their Ex Vivo Effects on Deformability, Indices, and Density of Circulating Erythrocytes in Patients Undergoing Cardiac Surgery With Cardiopulmonary.... J Cardiothorac Vasc Anesth. 2021; 36(3):855-861. DOI: 10.1053/j.jvca.2021.06.016. View

19.

Flegel W, Natanson C, Klein H . Does prolonged storage of red blood cells cause harm?. Br J Haematol. 2014; 165(1):3-16. PMC: 5515544. DOI: 10.1111/bjh.12747. View

20.

Cholette J, Pietropaoli A, Henrichs K, Alfieris G, Powers K, Gensini F . Elevated free hemoglobin and decreased haptoglobin levels are associated with adverse clinical outcomes, unfavorable physiologic measures, and altered inflammatory markers in pediatric cardiac surgery patients. Transfusion. 2018; 58(7):1631-1639. PMC: 6105435. DOI: 10.1111/trf.14601. View