Differential Expression Levels of Plasma microRNAs in Neuroblastoma Patients Identified by Next-Generation Sequencing

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  • ARFAN Ahmad
    Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan
  • KURIHARA Sho
    Department of Pediatric Surgery, Hiroshima University Hospital, Hiroshima University, Hiroshima, Japan
  • KOJIMA Masato
    Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan
  • KIMURA Shingo
    Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan
  • KANAWA Masami
    Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan
  • SOTOMARU Yusuke
    Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan
  • HIYAMA Eiso
    Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan Department of Pediatric Surgery, Hiroshima University Hospital, Hiroshima University, Hiroshima, Japan

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Abstract

<p>Efforts to identify biomarkers for neuroblastoma (NB) have been ongoing, but no definite biomarker has been identified in peripheral blood. We proposed the use of plasma exosomal miRNAs as biomarkers of unfavorable NB patient outcomes. Exosomal miRNAs isolated from 31 plasma and 37 tissue samples, many from the same NB patients, were sequenced using a next-generation sequencing instrument. We analyzed the correlation between miRNA expression levels in plasma and tissue samples with International Neuroblastoma Risk Group staging system (INRGSS) outcome and MYCN status. We chose differentially expressed miRNAs with similar expression patterns in plasma and tissue samples in each of the three analysis groups and combined those miRNAs to find the optimal combination with the potential to be considered as a biomarker. MicroRNA-92a-3p was found to be significantly upregulated in deceased patients (p = 0.017), miR-375 was upregulated in INRGSS stage M patients (p = 0.002), and plasma miR-92a-3p and miR-99a-5 levels were upregulated in patients with MYCN amplification (p = 0.007 and 0.006). The combination of miR-92a-3p, miR-375, and miR-99a-5p levels was shown to be a statistically significant predictor of NB patient outcomes (AUC = 0.726, p = 0.001, 95% CI = 0.612–0.841, sensitivity = 77%, specificity = 56.7%). Thus, the combination of miR-92a3p, miR-375, and miR-99a-5p may potentially be used as a biomarker for unfavorable NB patient outcomes. However, further validation is required in a larger number of NB patients.</p>

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