血小板浓缩物储存过程中的全转录组分析。
Whole transcriptome analysis of platelet concentrates during storage.
机构信息
Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Kashgar District Central Blood Station, Xinjiang Uygur Autonomous Region, Kashgar, China.
出版信息
Blood Transfus. 2023 Mar;21(2):146-156. doi: 10.2450/2022.0291-21. Epub 2022 Feb 16.
BACKGROUND
Platelets are anucleated blood cells and contain various RNA species. We investigated the changes in the whole transcriptome expression profile of platelet concentrates (PC) during storage to explore biological functions and biomarkers in platelet storage damage.
MATERIALS AND METHODS
Platelets were collected by apheresis from eight healthy blood donors and stored from day 0 to day 4. Platelet phenotyping and function analysis were used to detect platelet activity during storage. RNA-sequencing was used to detect changes in expression of mRNA, lncRNA and circRNA in the PC during storage. Gene ontology and KEGG analyses were applied to predict the functional distribution of differential expression of mRNA. Gene set enrichment analysis was used to analyze the differential levels of gene pathways. Finally, polymerase chain reaction (PCR) analysis was performed to verify the expression of three mRNA (POLE2, DCUN1D4, DAD1).
RESULTS
In total, 10,767 mRNA, 2,923 lncRNA and 68,550 circRNA were detected in the PC by RNA-sequencing. The expression levels of 222 mRNA changed significantly from day 0 to day 4 of storage: 58 increased continuously and 145 decreased continuously. Differentially expressed mRNA may be involved in physiological processes such as platelet activation, platelet aggregation, endocytosis, and apoptosis. Expression levels of 1,413 lncRNA were obvious. The levels of 42 species increased and the levels of 28 species decreased. The expression levels of 198 species of circRNA changed significantly, with those of 13 species changing continuously. The differential levels of expression of DAD1, DCUN1D4 and POLE1 mRNA, shown by RNA sequencing, were validated by PCR assay.
DISCUSSION
Changes in mRNA, lncRNA and circRNA during platelet storage may be closely related to platelet apoptosis and physiological functions in the platelet storage lesion. The expression levels of DAD1, DCUN1D4 and POLE1 could be biomarkers to monitor platelet status in PC bags.
背景
血小板是无核血细胞，含有各种 RNA 种类。我们研究了血小板浓缩物（PC）在储存过程中整个转录组表达谱的变化，以探索血小板储存损伤中的生物学功能和生物标志物。
材料与方法
从 8 名健康献血者中通过单采法采集血小板，从第 0 天到第 4 天储存。使用血小板表型和功能分析来检测储存过程中血小板的活性。使用 RNA 测序检测 PC 中 mRNA、lncRNA 和 circRNA 在储存过程中的表达变化。GO 和 KEGG 分析用于预测 mRNA 差异表达的功能分布。基因集富集分析用于分析基因通路的差异水平。最后，进行聚合酶链反应（PCR）分析以验证三种 mRNA（POLE2、DCUN1D4、DAD1）的表达。
结果
通过 RNA 测序共检测到 PC 中的 10767 个 mRNA、2923 个 lncRNA 和 68550 个 circRNA。从第 0 天到第 4 天储存时，222 个 mRNA 的表达水平发生显著变化：58 个持续增加，145 个持续减少。差异表达的 mRNA 可能参与血小板激活、血小板聚集、内吞作用和细胞凋亡等生理过程。1413 个 lncRNA 的表达水平明显。42 个物种的水平增加，28 个物种的水平减少。198 种 circRNA 的表达水平发生显著变化，其中 13 种持续变化。RNA 测序显示 DAD1、DCUN1D4 和 POLE1 mRNA 的差异水平表达通过 PCR 检测得到验证。
讨论
血小板储存过程中 mRNA、lncRNA 和 circRNA 的变化可能与血小板凋亡和血小板储存损伤中的生理功能密切相关。DAD1、DCUN1D4 和 POLE1 的表达水平可能是监测 PC 袋中血小板状态的生物标志物。
Zotero 插件