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High Autophagy Patterns in Swelling Platelets During Apheresis Platelet Storage

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Abstract

Platelets undergo remarkable morphological changes during storage. Platelets change into different sizes and densities and differ in their biochemistry and functions. However, the correlation between structural heterogeneity and platelet autophagy is largely unknown. The aim of this study was to investigate the autophagy process in vitro, such as routine storage of platelets, and explore the role of reactive oxygen species (ROS) involved in the regulation of platelet autophagy. The ROS and autophagy levels of platelet concentrates from apheresis platelets were evaluated through flow cytometry. The expression levels of autophagy-associated proteins (LC3I, LC3II, Beclin1, Parkin, and PINK1) were measured via Western blot. All biomarkers were dynamically monitored for seven days. Moreover, the morphological characteristics of platelet morphology during storage were analyzed through transmission electron microscopy (TEM). Flow cytometry showed that the levels of total cell ROS and mitochondria ROS increased in the stored platelets. Together with the increase in mitochondrial ROS, the autophagy signal LC3 in the platelets was strongly amplified. The number of swollen platelets (large platelets) considerably increased, and that of autophagy signal LC3 was remarkably higher than that of the normal platelets. Western blot revealed that the expression levels of Beclin1 and LC3 II/LC3 I ratio were enhanced, whereas those of Parkin and PINK1 almost did not change during the seven days of storage. The existence of autophagosomes or autophagolysosomes in the platelets at the middle stage of platelet storage was observed via TEM. Our data demonstrated that the subpopulation of large (swollen) platelets exhibited different autophagy patterns. Furthermore, increased platelet autophagy was associated with mitochondrial ROS. These preliminary results suggest that swelling platelets have a higher autophagy pattern than normal platelets during storage.

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Funding

We thank Yan Wang and Huihui Feng for contribution to serve the blood donors and PCs collection. This work was supported by grants from Natural Science Foundation of Ningbo (No.2019A610273 and No.2019A610268); Ningbo Public welfare science and technology project (No.2019C50083); Natural Science Foundation of Zhejiang Province (No.LY20H290006); Open Foundation of key laboratory of blood safety research of zhejiang province (No.2018KF010).

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Author notes

  1. Lu Yu and Shifang Yu authors contributed equally to this work.

Authors and Affiliations

  1. The Ningbo Central Blood Station, Ningbo, China

    Lu Yu, Yunlei He & Gang Deng

  2. The Department of Transfusion Medicine, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China

    Shifang Yu

  3. The Department of Laboratory Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China

    Qiang Li

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  1. Lu Yu
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Contributions

GD and QL conceived and designed the experiments; LY, SY and YH performed the experiments and evaluated the results; LY and QL wrote the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Qiang Li.

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Yu, L., Yu, S., He, Y. et al. High Autophagy Patterns in Swelling Platelets During Apheresis Platelet Storage. Indian J Hematol Blood Transfus 39, 670–678 (2023). https://doi.org/10.1007/s12288-023-01638-1

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