Circulating miR-26a as Potential Prognostic Biomarkers in Pediatric Rhabdomyosarcoma.

Angelica Zin,Beniamina Pacchioni,Caterina Millino,Lucia Tombolan,Paolo Bonvini,Gianni Bisogno,Manuela Cattelan

doi:10.3389/fgene.2020.606274

Angelica Zin, Beniamina Pacchioni + Show 5 more

Open Access

https://doi.org/10.3389/fgene.2020.606274

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Abstract

Rhabdomyosarcoma (RMS) arises from myogenic precursors that fail to complete muscle differentiation and represents the most frequent soft tissue sarcoma in children. Two major histological subtypes are recognized: alveolar RMS, characterized by a more aggressive behavior and a greater proneness to metastasis, and embryonal RMS which accounts for the 80% of cases and carries a better prognosis. Despite the survival of patients with localized tumors has progressively improved, RMS remains a challenging disease especially for metastatic patients and in case of progressive or recurrent disease after front-line therapy. MicroRNAs, a class of small non-coding RNA, have emerged as crucial players in cancer development and progression, and their detection in plasma (circulating miRNAs) represents a promising minimally invasive approach that deserve to be exploited in clinical practice. We evaluated the utility of circulating miRNAs as diagnostic and prognostic biomarkers in children with RMS profiling miRNAs from plasma of a small cohort of RMS patients and healthy donors (HD) using a qPCR Cancer Panel. An assessment of hemolysis status of plasma using miR-451/miR-23a ratio was performed as pre-analytical analysis. Statistical analysis revealed that miRNAs expression pattern clearly distinguished RMS patients from HD (p < 0.05). Interestingly, plasma levels of muscle-specific miR-206 were found to be significantly increased in RMS patients compared to HD, whereas levels of three potential tumor-suppressor miRNAs, miR-26a and miR-30b/30c, were found lower. Reduced levels of circulating miR-26a and miR-30b/c were further measured in an independent larger cohort of patients (validation set) by digital droplet PCR. In particular, we evidenced that miR-26a absolute plasma levels were associated with fusion status and adverse outcome (p < 0.05). Taken together, these findings demonstrate the potential of circulating miRNA as diagnostic and prognostic biomarker in children affected by this malignancy and enforced the key role of miR-26a in pediatric rhabdomyosarcoma.

Highlights

Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma affecting children
We applied the Quantitative PCR (qPCR) Cancer Panel to a training group of plasma samples composed of 8 RMS patients
The Volcano plot displayed a good distribution of normalized data using either one of the two normalization approaches, and when unsupervised hierarchical clustering analysis was performed using all miRNAs of the qPCR Cancer Panel a good separation between groups (ARMS, embryonal RMS (ERMS), and healthy donors (HD)) was observed (Supplementary Figure 1C)

Summary

Introduction

It arises from myogenic precursor that are unable to complete muscle differentiation Typical myogenic factors such as MyoD1 and myogenin are expressed by tumor cells and routinely used to define the diagnosis of RMS (Sebire and Malone, 2003). About 80% of ARMS are characterized by a reciprocal translocation, the more frequent t (2;13) and the less common t (1;13) (Rudzinski et al, 2015). Both translocations involve PAX genes, PAX3 and PAX7 respectively, and the transcription factor FOXO1. Fusion-positive ARMS tumors are associated with a more aggressive phenotype and worse prognosis respect to the embryonal RMS(Sorensen et al, 2002). Detection of relapse in children with RMS includes bone marrow aspirate and biopsy and computed tomography (CT) scan (Huh and Skapek, 2010)

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