Date: Saturday, May 30, 2020
Session Time: 3:15pm-4:00pm
Presentation Time: 3:30pm-4:00pm
*Purpose: Donor quality plays a critical role during IRI and early graft outcomes. Extracellular vesicles (EVs) provide a novel source of valuable biomarkers that may reflect organ quality and allograft function-post-transplant.
*Methods: We evaluated perfusate (10 cc) from 70 deceased and live liver grafts obtained at the end of cold ischemia time (CIT) Our studies include determination of EV size(Tunable Resistive Pulse Sensing (TRPS) and Cryo- Electron Microscopy (CEM)), quantification (using TRPS and Imaging Flow Cytometry (IFC)) and phenotyping (using IFC).Deceased donor (DD) livers included standard, extended criteria donor as well as partial (living donor (LD)) grafts. EV-miRNAs were evaluated using miScript miRNA PCR Array Human Liver miFinder 384H. For identification of differentially expressed EV-miRNAs the ΔΔCt method was used. CIT, donor age, and transaminase levels at day 1 and 2 post-transplant were included as part of the analyses.
*Results: Results: Using a volume of 200 cc of perfusate, we were able to quantify ~ 1.5E+08 EVs. The effect of CIT on perfusate EV-miRNA profiles from DD (CIT > 4 hours) and LD organs (CIT < 1 hour) showed increase of miR-5701 and a decrease of miR-122-5p, miR-260a. miR-19a- 3p, miR-3183, miR-4301, and miR-92a-3p (Fig. 1). Older donors livers (> 65 years old) showed a significant up regulation of miR 122-3p, miR-122-5p, miR-1260 and miR-192-5p in their perfusates compared to young donor livers. Furthermore, a significant increase of AST/ALT on recipient at 24 hrs post-transplantation relates to increased expression of miR-101-3p, miR-106b-5p, miiR-142-3p, miR-199b-3p, miR-22-3p, miR-29a-3p, miR-29b-3p, and miR-29c-3p (Fig. 2). Critically, a miRNA signature was identified associated with donor quality.
*Conclusions: Perfusate EV-miRs are excellent candidates as markers of donor quality and tissue injury and the profiles relates to early graft outcomes. Further evaluation of cellular origin affected pathways may lead to discovery of target for therapeutic interventions.
To cite this abstract in AMA style:Bajwa A, Kuscu C, Kuscu C, Wolen AR, Eason JD, Maluf D, Mas V. Small RNA Extracellular Vesicles Isolated from Liver Perfusate Provides Assessment of Organ Quality and Post Transplantation Graft Function [abstract]. Am J Transplant. 2020; 20 (suppl 3). https://atcmeetingabstracts.com/abstract/small-rna-extracellular-vesicles-isolated-from-liver-perfusate-provides-assessment-of-organ-quality-and-post-transplantation-graft-function/. Accessed March 4, 2021.
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