Department of Genetics, Ribeirao Preto Medical School, University of Sao Paulo, Ribeirao Preto, Brazil.
Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of Sao Paulo, Pirassununga, Sao Paulo, Brazil.
Sci Rep. 2017 Oct 30;7(1):14319. doi: 10.1038/s41598-017-14616-1.
Nuclear reprogramming mediated by somatic cell nuclear transfer (SCNT) has many applications in medicine. However, animal clones show increased rates of abortion and reduced neonatal viability. Herein, we used exosomal-miRNA profiles as a non-invasive biomarker to identify pathological pregnancies. MiRNAs play important roles in cellular proliferation and differentiation during early mammalian development. Thus, the aim of this study was to identify exosomal-miRNAs in maternal blood at 21 days of gestation that could be used for diagnosis and prognosis during early clone pregnancies in cattle. Out of 40 bovine-specific miRNAs, 27 (67.5%) were with low abundance in the C-EPL (Clone - Early pregnancy loss) group compared with the C-LTP (Clone - Late pregnancy) and AI-LTP (Artificial Insemination - Late pregnancy) groups, which had similar miRNAs levels. Bioinformatics analysis of the predicted target genes demonstrated signaling pathways and functional annotation clusters associated with critical biological processes including cell proliferation, differentiation, apoptosis, angiogenesis and embryonic development. In conclusion, our results demonstrate decreased exosomal-miRNAs in maternal blood at 21 days of gestation in cloned cattle pregnancies that failed to reach term. Furthermore, the predicted target genes regulated by these 27 miRNAs are strongly associated with pregnancy establishment and in utero embryonic development.
体细胞核移植(SCNT)介导的核重编程在医学中有许多应用。然而,动物克隆显示出流产率增加和新生仔畜活力降低的情况。在此,我们使用外泌体-miRNA 谱作为一种非侵入性生物标志物来识别病理性妊娠。miRNA 在哺乳动物早期发育过程中的细胞增殖和分化中发挥重要作用。因此,本研究旨在鉴定在牛早期克隆妊娠中,妊娠 21 天时母血中的外泌体-miRNA,用于诊断和预后。在 40 种牛特异性 miRNA 中,与 C-LTP(克隆-晚期妊娠)和 AI-LTP(人工授精-晚期妊娠)组相比,在 C-EPL(克隆-早期妊娠丢失)组中 27 种(67.5%)miRNA 的丰度较低,这两组的 miRNA 水平相似。对预测靶基因的生物信息学分析表明,与细胞增殖、分化、凋亡、血管生成和胚胎发育等关键生物学过程相关的信号通路和功能注释簇。总之,我们的研究结果表明,在未能达到足月的克隆牛妊娠中,妊娠 21 天时母血中外泌体-miRNA 减少。此外,这 27 个 miRNA 调控的预测靶基因与妊娠建立和宫内胚胎发育密切相关。