Cold storage conditions modify microRNA expressions for platelet transfusion
Cold storage conditions modify microRNA expressions for platelet transfusion
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DOI：
10.1371/journal.pone.0218797
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发表时间：
2019-07
期刊：
影响因子：
3.7
通讯作者：
N. Mukai;Yoshinobu Nakayama;Sachiyo Ishi;Takayuki Murakami;S. Ogawa;K. Kageyama;S. Murakami;Yuji Sasada;J. Yoshioka;Y. Nakajima
中科院分区：
文献类型：
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作者：
N. Mukai;Yoshinobu Nakayama;Sachiyo Ishi;Takayuki Murakami;S. Ogawa;K. Kageyama;S. Murakami;Yuji Sasada;J. Yoshioka;Y. Nakajima
MicroRNAs （miRNAs）是在造血过程中调节基因和蛋白表达的小RNA分子。众所周知，血小板含有功能齐全的miRNA机制。虽然用于输血的血小板通常在室温下储存，但最近的证据表明，在冷藏条件下更有利，包括更高的粘附性和聚集性。因此，我们试图确定血小板的功能差异是否与血小板miRNA表达的差异谱有关。为了获得miRNA表达谱，对10名健康受试者的人血小板进行了新一代测序。在三种不同的条件下储存mirna: 1)基线（储存前），2)在22°C下搅拌72 h， 3)在4°C下储存72 h。通过测序鉴定mirna后，使用定量聚合酶链反应（qPCR）在18名健康受试者的成熟mirna水平上验证结果。与基线相比，在4°C保存的125个mirna和在22°C保存的9个mirna存在差异表达。qPCR验证研究证实，与22°C保存的样品相比，4°C保存的样品中mir-20a-5p （1.87, p<0.0001）、mir-10a-3p （1.88, p<0.0001）、mir-16-2-3p （1.54, p<0.01）和mir-223-5p （1.38, p<0.05）的表达水平（fold change 95% CI）增加。这些结果表明，在特定的储存条件下，mirna与血小板质量相关。这些数据表明，mirna可能被用作血小板质量的生物标志物。
MicroRNAs (miRNAs) are small RNA molecules that modulate gene and protein expression in hematopoiesis. Platelets are known to contain a fully functional miRNA machinery. While platelets used for transfusion are normally stored at room temperature, recent evidence suggests more favorable effects under a cold-storage condition, including higher adhesion and aggregation properties. Thus, we sought to determine whether functional differences in platelets are associated with the differential profiling of platelet miRNA expressions. To obtain the miRNA expression profile, next-generation sequencing was performed on human platelets obtained from 10 healthy subjects. The miRNAs were quantified after being stored in three different conditions: 1) baseline (before storage), 2) stored at 22°C with agitation for 72 h, and 3) stored at 4°C for 72 h. Following the identification of miRNAs by sequencing, the results were validated at the level of mature miRNAs from 18 healthy subjects, by using quantitative polymerase chain reaction (qPCR). Differential expression was observed for 125 miRNAs that were stored at 4°C and 9 miRNAs stored at 22°C as compared to the baseline. The validation study by qPCR confirmed that storage at 4°C increased the expression levels (fold change 95% CI) of mir-20a-5p (1.87, p<0.0001), mir-10a-3p (1.88, p<0.0001), mir-16-2-3p (1.54, p<0.01), and mir-223-5p (1.38, p<0.05), compared with those of the samples stored at 22°C. These results show that miRNAs correlate with platelet quality under specific storage conditions. The data indicate that miRNAs could be potentially used as biomarkers of platelet quality.