Hsa-miR-3651 could serve as a novel predictor for in-breast recurrence via FRMD3
PO-1450
Abstract
Hsa-miR-3651 could serve as a novel predictor for in-breast recurrence via FRMD3
Authors: Elvis Ruznic1, Barbara Zellinger2,3, Ulrich Bodenhofer4,5, Immanuela A. Engländer6,6, Cornelia Kronberger3, Brane Grambozov1, Markus Stana1, Josef Karner1, Gerd Fastner1, Karl Sotlar3, Felix Sedlmayer1,1, Franz Zehentmayr1,6
1Paracelsus Medical University Salzburg, Department of Radiation Oncology, Salzburg, Austria; 2Paracelsus Medical University Salzburg, radART - Institute for Research and Development on Advanced Radiation Technologies, Salzburg, Austria; 3Paracelsus Medical University Salzburg, Department of Pathology, Salzburg, Austria; 4University of Applied Sciences Upper Austria, School of Informatics, Communications and Media, Hagenberg, Austria; 5Johannes Kepler University, Institute for Machine Learning, Linz, Austria; 6Paracelsus Medical University Salzburg, radART—Institute for Research and Development on Advanced Radiation Technologies, Salzburg, Austria
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Purpose or Objective
MicroRNAs are small non-coding RNAs with pivotal regulatory functions in multiple cellular processes. Their significance as molecular predictors for breast cancer was demonstrated in the past 15 years. The aim of this study was to elucidate the role of hsa-miR-3651 for predicting of local control (LC) in early breast cancer.
Material and Methods
The pilot phase of this study consisted of two high-throughput experiments. In the first run, we screened formalin-fixed paraffin-embedded (FFPE) breast cancer specimens for microRNAs using microarrays (Agilent®). Secondly, we used Affymetrix GeneChips® to screen for target proteins in the same tissue samples. Subsequently, an in-silico analysis was conducted to detect differentially expressed microRNAs and target genes using Linear Models for Microarray Data (LIMMA). Additionally, we compared potential targets with publicly available databases (TargetScan, miRDB, PITA, DIANA, DIANA Cancer, miRCarta). In the validation phase of this study, these results were validated with RT-qPCR (microRNA) as well as droplet digital PCR (target proteins) in an independent patient cohort.
Results
Hsa-miR-3651 was found to be differentially expressed between patients who experienced local relapse compared to those without (N = 23; p = 0.0035). This result could be validated in an independent cohort of 87 patients using RT-qPCR (p < 0.0005). In a second step FERM domain protein 3 (FRMD3) was found to be the most down-regulated protein (N = 21; p = 0.0016). Computational analysis employing different prediction algorithms revealed FRMD3 as a likely downstream target of hsa-miR-3651 with a 8mer binding site between the two molecules, which could be corroborated by droplet digital PCR in an independent patient set (N = 20, p = 0.134).
Conclusion
The current study revealed that hsa-miR-3651 might be a predictor of LC in early breast cancer via its putative target protein FRMD3. Since microRNAs interfere in multiple pathways, the results of this hypothesis generating study may contribute to the development of tailored therapies for breast cancer in the future.