Pathogen inactivation with amotosalen plus UVA illumination minimally impacts microRNA expression in platelets during storage under standard blood banking conditions

Abstract

BACKGROUND: To reduce the risk of transfusion transmission infection, nucleic acid targeted methods have been developed to inactivate pathogens in PCs. miRNAs have been shown to play an important role in platelet function, and changes in the abundance of specific miRNAs during storage have been observed, as have perturbation effects related to pathogen inactivation (PI) methods. The aim of this work was to investigate the effects of PI on selected miRNAs during storage.STUDY DESIGN AND METHODS: Using a pool and split strategy, 3 identical buffy coat PC units were generated from a pool of 24 whole blood donors. Each unit received a different treatment: 1) Untreated platelet control in platelet additive solution (C-PAS); 2) Amotosalen-UVA-treated platelets in PAS (PI-PAS); and 3) untreated platelets in donor plasma (U-PL). PCs were stored for 7 days under standard blood banking conditions. Standard platelet quality control (QC) parameters and 25 selected miRNAs were analyzed.RESULTS: During the 7-day storage period, differences were found in several QC parameters relating to PI treatment and storage in plasma, but overall the three treatments were comparable. Out of 25 miRNA tested changes in regulation of 5 miRNA in PI-PAS and 3 miRNA U-PL where detected compared to C-PAS. A statistically significant difference was observed in down regulations miR-96-5p on Days 2 and 4, 61.9% and 61.8%, respectively, in the PI-PAS treatment. CONCLUSION:Amotosalen-UVA treatment does not significantly alter the miRNA profile of platelet concentrates generated and stored using standard blood banking conditions. P latelets play a central role in hemostasis and represent an integral part of transfusion medicine. Platelet concentrates (PCs) are normally stored at room temperature for a maximum of 5 to 7 days, depending on country-specific regulations. 1,2 One of two main reasons for this limited storage time is the potential for bacterial growth during storage. 3,4 The risk of transfusion-transmitted bacterial infections (TTBIs) due to contaminated PCs is persistent despite improved donor selection protocols, phlebotomy techniques, and PC bacterial screening. Life threatening septic transfusion reactions (STRs) related to platelet transfusions still occur. 4,5 The currently available bacterial detection assays are not sufficient to prevent TTBI. 6 The rate of STRs due to PC transfusion has likely been underestimated, as highlighted in a recent 7-year retrospective study of PC transfusions using passive and active surveillance. 7 The second reason for the relatively short shelf life of stored platelets is platelet storage lesion (PSL). PSL is a collective term for a variety of factors that describe the deterioration of platelet quality during storage. 8,9 The onset and acceleration of PSL during storage is likely to affect platelet post transfusion recovery and survival. 10,11 The onset and accumulation of PSL is related to the activation of or damage to the platelets as result of their preparation or storage conditions, and can eventual...

Protein Concentrations in Stored Pooled Platelet Concentrates Treated with Pathogen Inactivation by Amotosalen Plus Ultraviolet a Illumination

“…Using various markers of platelet activation and granule release, a number of studies have noted accelerated PSL during storage in PI-treated PCs compared to standard PCs [ 23 , 73 , 74 ]. In our previously published data [ 80 ], we also saw some indications of this effect, with increased shedding of the GPIba receptor and surface expression of P-selectin. The molecular basis for increased PSL during storage and the potential effects of additional processing like PI treatment have been extensively studied in recent years, though there have been conflicting results and debate on their clinical relevance.…”

“…There is some evidence of miRNA regulation of de-granulation. In our previous published data on miRNA in platelets during storage, miRNA-96 was downregulated in platelets treated with PI [ 80 ]. miRNA-96 has been implicated in regulation of vesicle-associated membrane protein 8 (VAMP8) that effects the degranulation of platelets [ 81 , 82 ].…”

Ultraviolet-Based Pathogen Inactivation Systems: Untangling the Molecular Targets Activated in Platelets

“…Further, the reduction in the platelet miRNA levels induced by INTERCEPT correlated with platelet activation and an impaired platelet aggregation response to ADP ( 136 ). In contrast, a recent study presented by Arnason et al ( 138 ) demonstrated that INTERCEPT treatment did not change the quality or significantly altered the miRNA profile of PCs. These controversial results prompted further investigations and as the clinical significance of MV-associated miRNAs is unknown, and speculation of a negative effect of PI-treated platelets including long-term consequences for recipients is as yet unwarranted.…”

Inactivation of three emerging viruses – severe acute respiratory syndrome coronavirus, Crimean–Congo haemorrhagic fever virus and Nipah virus – in platelet concentrates by ultraviolet C light and in plasma by methylene blue plus visible light

“…Both pathogen inactivation systems have been tested in vitro to be effective against many different types of viruses, including emerging viruses such as West Nile virus and yellow fever virus . In this study, we investigated the capacity of THERAFLEX UV‐Platelets and THERAFLEX MB‐Plasma systems to inactivate the emerging viruses SARS‐CoV, CCHFV and NiV in PCs and plasma, respectively.…”