PMID- 32928548 OWN - NLM STAT- MEDLINE DCOM- 20210802 LR - 20210802 IS - 1097-685X (Electronic) IS - 0022-5223 (Linking) VI - 162 IP - 2 DP - 2021 Aug TI - Cell-free DNA in human ex vivo lung perfusate as a potential biomarker to predict the risk of primary graft dysfunction in lung transplantation. PG - 490-499.e2 LID - S0022-5223(20)32354-0 [pii] LID - 10.1016/j.jtcvs.2020.08.008 [doi] AB - BACKGROUND: Cell-free DNA (cfDNA), such as mitochondrial DNA (mtDNA) and nuclear DNA (nuDNA), are known to be released from injured cells and as such have been explored as biomarkers for tissue injury in different clinical settings. Ex vivo lung perfusion (EVLP) has been developed as an effective technique for marginal donor lung functional assessment. We hypothesized that the level of cfDNA in EVLP perfusate may reflect tissue injury and thus can be developed as a biomarker to quantify the degree of donor lung injury or predict the development of primary graft dysfunction (PGD) after lung transplantation (LTx). METHODS: The perfusate from 62 donor lungs transplanted at our institution between May 2010 and December 2015 was sampled for cfDNA at 1 and 4 hours of perfusion. Sequences of genes encoding nicotinamide adenine dinucleotide dehydrogenase 1 (NADH-1) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used to represent mtDNA and nuDNA, respectively. Levels were quantified by real-time polymerase chain reaction and correlated with clinical outcome after LTx. RESULTS: In our entire cohort, 14 patients developed PGD grade 3 (PGD3) within 72 hours after LTx. The non-PGD group included 48 patients (PGD0-1). Concentrations of mtDNA in the perfusate of the PGD3 group were significantly higher than those in non-PGD group at 1 hour of EVLP (1874 +/- 844 vs 1259 +/- 885 copies/muL; P = .011). The perfusate of the PGD3 group had significantly higher levels of nuDNA compared with the non-PGD group at both 1 hour (1498 +/- 1895 vs 675 +/- 391 copies/muL; P = .008) and 4 hours (4521 +/- 5810 vs 1764 +/- 1494 copies/muL; P = .001). In donation after cardiac death (DCD) cases, mtDNA levels were significantly higher in the PGD3 group compared with the non-PGD group at 1 hour of EVLP (2060 +/- 997 vs 1184 +/- 782 copies/muL; P = .040), and the levels of nuDNA were significantly higher in the PGD3 group compared with the non-PGD group at both 1 hour (1021 +/- 495 vs 606 +/- 305 copies/muL; P = .041) and 4 hours (2358 +/- 1028 vs 1185 +/- 967 copies/muL; P = .006). In donation after brain death (DBD) cases, cfDNA scores did not show a significant difference. CONCLUSIONS: We found that the amount of cfDNA, especially nuDNA, in EVLP perfusate was higher in the severe PGD group (PGD3) compared with the non-PGD group. This proof-of-concept study supports the concept that the analysis of cfDNA levels in EVLP perfusate can help estimate the damage to donor lungs before implantation. Larger studies are needed to validate this concept. CI - Copyright (c) 2020. Published by Elsevier Inc. FAU - Kanou, Takashi AU - Kanou T AD - Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada. FAU - Nakahira, Kiichi AU - Nakahira K AD - Division of Pulmonary and Critical Care Medicine, Weill Cornell Medicine, New York, NY. FAU - Choi, Augustine M AU - Choi AM AD - Division of Pulmonary and Critical Care Medicine, Weill Cornell Medicine, New York, NY. FAU - Yeung, Jonathan C AU - Yeung JC AD - Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada. FAU - Cypel, Marcelo AU - Cypel M AD - Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada. FAU - Liu, Mingyao AU - Liu M AD - Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada. FAU - Keshavjee, Shaf AU - Keshavjee S AD - Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto, Ontario, Canada. Electronic address: shaf.keshavjee@uhn.ca. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Video-Audio Media DEP - 20200811 PL - United States TA - J Thorac Cardiovasc Surg JT - The Journal of thoracic and cardiovascular surgery JID - 0376343 RN - 0 (Biomarkers) RN - 0 (Cell-Free Nucleic Acids) SB - IM CIN - J Thorac Cardiovasc Surg. 2021 Aug;162(2):501-502. PMID: 32888703 CIN - J Thorac Cardiovasc Surg. 2021 Aug;162(2):500. PMID: 32981702 CIN - J Thorac Cardiovasc Surg. 2021 Aug;162(2):512-513. PMID: 33618880 MH - Adult MH - Biomarkers/analysis MH - Cell-Free Nucleic Acids/*analysis MH - Female MH - Humans MH - Lung Transplantation/*adverse effects MH - Male MH - Middle Aged MH - *Perfusion MH - Predictive Value of Tests MH - Primary Graft Dysfunction/diagnosis/*etiology MH - Proof of Concept Study MH - Retrospective Studies MH - Risk Assessment MH - Risk Factors MH - Severity of Illness Index MH - Time Factors MH - Treatment Outcome OTO - NOTNLM OT - cell-free DNA OT - ex vivo lung perfusion OT - lung transplantation OT - primary graft dysfunction EDAT- 2020/09/16 06:00 MHDA- 2021/08/03 06:00 CRDT- 2020/09/15 05:39 PHST- 2019/12/27 00:00 [received] PHST- 2020/07/10 00:00 [revised] PHST- 2020/08/04 00:00 [accepted] PHST- 2020/09/16 06:00 [pubmed] PHST- 2021/08/03 06:00 [medline] PHST- 2020/09/15 05:39 [entrez] AID - S0022-5223(20)32354-0 [pii] AID - 10.1016/j.jtcvs.2020.08.008 [doi] PST - ppublish SO - J Thorac Cardiovasc Surg. 2021 Aug;162(2):490-499.e2. doi: 10.1016/j.jtcvs.2020.08.008. Epub 2020 Aug 11.