Proteomic analysis of machine perfusion solution from brain dead donor kidneys reveals that elevated complement, cytoskeleton and lipid metabolism proteins are associated with 1‐year outcome

Abstract

SUMMARY Assessment of donor kidney quality is based on clinical scores or requires biopsies for histological assessment. Noninvasive strategies to identify and predict graft outcome at an early stage are, therefore, needed. We evaluated the perfusate of donation after brain death (DBD) kidneys during nonoxygenated hypothermic machine perfusion (HMP). In particular, we compared perfusate protein profiles of good outcome (GO) and suboptimal outcome (SO) 1‐year post‐transplantation. Samples taken 15 min after the start HMP (T1) and before the termination of HMP (T2) were analysed using quantitative liquid chromatography–tandem mass spectrometry (LC‐MS/MS). Hierarchical clustering of the 100 most abundant proteins showed discrimination between grafts with a GO and SO at T1. Elevated levels of proteins involved in classical complement cascades at both T1 and T2 and a reduced abundance of lipid metabolism at T1 and of cytoskeletal proteins at T2 in GO versus SO was observed. ATP‐citrate synthase and fatty acid‐binding protein 5 (T1) and immunoglobulin heavy variable 2‐26 and desmoplakin (T2) showed 91% and 86% predictive values, respectively, for transplant outcome. Taken together, DBD kidney HMP perfusate profiles can distinguish between outcome 1‐year post‐transplantation. Furthermore, it provides insights into mechanisms that could play a role in post‐transplant outcomes.

Doxycycline Alters the Porcine Renal Proteome and Degradome during Hypothermic Machine Perfusion

“…This could indicate that there is detachment of remnant classical complement attack complexes in the doxycycline group. Interestingly, our group has shown that upregulation of C1s secretion during hypothermic machine perfusion correlates with a good kidney function 1-year post transplantation [47]. Furthermore, we identified significant downregulation of C3 degradation products before reperfusion and significant upregulation C3 degradation products after reperfusion (Figure 3b,c).…”

Shifting Paradigms for Suppressing Fibrosis in Kidney Transplants: Supplementing Perfusion Solutions With Anti-fibrotic Drugs

“…Last but not least, HMP allows for sampling of the perfusate to assess the viability, as elegantly shown by Moers et al ( 68 ) who analyzed glutathione-S-transferase, N-acetyl-β-D-glucosaminidase, and heart-type fatty acid binding protein to predict DGF. Emerging techniques, such as proteomics, also appear promising, as reported by van Leeuwen et al ( 69 ), who found out that the HMP perfusate composition associates with the 1-year transplant outcome.…”

A Review of Current and Emerging Trends in Donor Graft-Quality Assessment Techniques

“…Perfusate analysis at T1 revealed a predictive value of 91% for ATP-citrate synthase and fatty acid-binding protein 5, and analysis at T2 showed a predictive value of 86% for immunoglobulin heavy variable 2-26 and desmoplakin. In summary, HMP perfusate profiles for DBD kidneys can distinguish between outcomes one-year post-transplantation, providing a potential non-invasive method of assessing donor organ quality [2].…”

“…Unfortunately, the number of patients placed on kidney transplant waiting lists is rapidly increasing, resulting in a growing gap between organ demand and the availability of kidneys for transplantation. Standard criteria donors (SCD) are preferred for kidney transplants because organs from these individuals typically result in more favourable outcomes compared to other donor types [2]. However, the shortage of available kidneys has forced medical professionals to utilize marginal kidneys from expanded criteria donors (ECD) to broaden the donor pool and shorten wait times for patients with end-stage renal disease.…”

“…However, the shortage of available kidneys has forced medical professionals to utilize marginal kidneys from expanded criteria donors (ECD) to broaden the donor pool and shorten wait times for patients with end-stage renal disease. Nonetheless, it is well known that donor organ quality affects long-term outcomes for renal transplant recipients, and ECD kidney grafts have been shown to have worse outcomes compared to SCD grafts, including an increased risk of delayed graft function (DGF) and primary nonfunction incidence (PNF) [2,3]. Thus, representative methods of assessing graft-quality are urgently needed, especially for ECDs.…”