Medical Research Unit in Infectious Diseases, Hospital de Pediatría, CMN SXXI, Instituto Mexicano del Seguro Social, Mexico City, Mexico.
Epidemiology Department, Columbia University, New York, New York, United States of America.
PLoS One. 2020 Apr 14;15(4):e0231394. doi: 10.1371/journal.pone.0231394. eCollection 2020.
miRNAs regulate post-transcriptional gene expression in metazoans, and thus are involved in many fundamental cellular biological processes. Extracellular miRNAs are also found in most human biofluids including plasma. These circulating miRNAs constitute a long distance inter cellular communication system and are potentially useful biomarkers. High throughput technologies like microarrays are able to scan a complete miRNome providing useful detection scores that are underexplored. We proposed to answer how many and which miRNAs are detectable in plasma or extracellular vesicles as these questions have not yet been answered. We set out to address this knowledge gap by analyzing the mirRNome in plasma and corresponding extracellular vesicles (EVs) from 12 children affected by retinoblastoma (Rb) a childhood intraocular malignant tumor, as well as from 12 healthy similarly aged controls. We calculated an average of 537 detectable miRNAs in plasma and 625 in EVs. The most miRNA enriched compartment were EVs from Rb cases with an average of 656 detectable elements. Using hierarchical clustering with the detection scores, we generated broad detection mirnome maps and identified a plasma signature of 19 miRNAs present in all Rb cases that is able to discriminate cases from controls. An additional 9 miRNAs were detected in all the samples; within this group, miRNA-5787 and miRNA-6732-5p were highly abundant and displayed very low variance across all the samples, suggesting both are good candidates to serve as plasma references or normalizers. Further exploration considering participant's sex, allowed discovering 5 miRNAs which corresponded only to females and 4 miRNAs corresponding only to males. Target and pathway analysis of these miRNAs revealed hormonal function including estrogen, thyroid signaling pathways and testosterone biosynthesis. This approach allows a comprehensive unbiased survey of a circulating miRNome landscape, creating the possibility to define normality in mirnomic profiles, and to locate where in these miRNome profiles promising and potentially useful circulating miRNA signatures can be found.
miRNAs 调控后生转录基因表达，参与许多基本的细胞生物学过程。细胞外 miRNAs 也存在于大多数人体生物体液中，包括血浆。这些循环 miRNAs 构成了长距离细胞间通讯系统，具有潜在的应用价值。高通量技术，如微阵列，能够扫描完整的 miRNome，提供未被充分探索的有用检测评分。我们提出了一个问题，即在血浆或细胞外囊泡（EVs）中可以检测到多少和哪些 miRNAs，因为这些问题尚未得到解答。我们通过分析 12 名患有视网膜母细胞瘤（Rb）的儿童和 12 名年龄相仿的健康对照儿童的血浆和相应的细胞外囊泡（EVs）中的 mirRNome，来解决这一知识空白。我们计算出血浆中平均有 537 个可检测的 miRNAs，EVs 中有 625 个。miRNAs 最丰富的区室是 Rb 病例的 EVs，平均有 656 个可检测的元素。我们使用检测评分的层次聚类，生成了广泛的检测 mirnome 图谱，并鉴定出了一个存在于所有 Rb 病例中的 19 个 miRNA 的血浆特征，该特征能够区分病例和对照。在所有样本中还检测到另外 9 个 miRNA；在这个组中，miRNA-5787 和 miRNA-6732-5p 含量非常丰富，在所有样本中变异很小，这表明它们都是作为血浆参考或标准化物的良好候选物。进一步考虑参与者的性别进行探索，发现了 5 个只对应于女性的 miRNA 和 4 个只对应于男性的 miRNA。这些 miRNA 的靶标和通路分析显示了激素功能，包括雌激素、甲状腺信号通路和睾酮生物合成。这种方法允许对循环 miRNome 图谱进行全面、无偏的调查，从而有可能定义 miRNome 图谱的正常性，并确定在这些 miRNome 图谱中可以找到有希望和潜在有用的循环 miRNA 特征的位置。
