Section of Cell Biology, Laboratory of Cellular Hematology, Center for Biologics Evaluation and Research, FDA, Bethesda, Maryland, USA.
Transfusion. 2010 Jul;50(7):1581-8. doi: 10.1111/j.1537-2995.2010.02585.x. Epub 2010 Feb 11.
MicroRNAs (miRNAs), the negative regulators of cellular mRNAs, are present in mature red blood cells (RBCs) in abundance relative to other blood cells. So far, there are no studies aimed at identifying large-scale miRNA profiles during storage of RBCs.
RNA samples from each RBC bag stored at 4 degrees C were collected on Days 0, 20, and 40 and subjected to miRNA profiling by using a membrane-based array. Fifty-two selected miRNAs of cellular apoptotic pathway represent the array. Through bioinformatics analyses, we identified potential target genes for selected miRNAs.
Differential profiling of RBCs for 52 miRNAs revealed two distinguishable patterns during storage: Forty-eight miRNAs demonstrated no trend at all, while four miRNAs, miR-96, miR-150, miR-196a, and miR-197, demonstrated an increase up to Day 20 and subsequently decreased during storage. We selected miR-96 and subjected it to standard bioinformatics analyses for target gene predictions, which identified several mRNAs including the RBC proapoptotic calpain small subunit-1 (CAPNS1) as potential targets of miR-96. To validate these predictions, we selected CAPNS1 mRNA as an example and confirmed its presence in the RBCs. Future experimental verification would help define miR-96-CAPNS1 interaction, if any, in the stored RBCs.
This study for the first time provided a differential profile of stored RBCs for selected miRNAs related to cellular apoptotic pathway and opened new avenues toward identification of novel in vitro RBC biomarkers of storage lesions. Future studies focusing on target gene-miRNA interactions in stored RBCs would also unravel underlying mechanisms of storage lesions.
微小 RNA（miRNA）是细胞 mRNA 的负调控因子，在成熟的红细胞（RBC）中相对于其他血细胞大量存在。到目前为止，还没有研究旨在确定 RBC 储存过程中的大规模 miRNA 图谱。
在 4°C 下储存的每个 RBC 袋的 RNA 样本在第 0、20 和 40 天收集，并通过膜基阵列进行 miRNA 谱分析。该阵列包含细胞凋亡途径的 52 个选定 miRNA。通过生物信息学分析，我们确定了选定 miRNA 的潜在靶基因。
对 52 个 miRNA 的 RBC 进行差异谱分析显示，在储存过程中存在两种可区分的模式：48 个 miRNA 没有任何趋势，而 4 个 miRNA，miR-96、miR-150、miR-196a 和 miR-197，在储存过程中先增加到第 20 天，然后减少。我们选择 miR-96 并对其进行标准生物信息学分析以预测靶基因，该分析鉴定了几个 mRNA，包括 RBC 促凋亡钙蛋白酶小亚基-1（CAPNS1）作为 miR-96 的潜在靶标。为了验证这些预测，我们选择 CAPNS1 mRNA 作为示例并确认其在 RBC 中存在。未来的实验验证将有助于确定 miR-96-CAPNS1 相互作用，如果有的话，在储存的 RBC 中。
本研究首次提供了与细胞凋亡途径相关的储存 RBC 选定 miRNA 的差异图谱，并为鉴定储存损伤的新型 RBC 体外生物标志物开辟了新途径。未来专注于储存 RBC 中靶基因-miRNA 相互作用的研究也将揭示储存损伤的潜在机制。
