Cross‐species identification of a plasma microRNA signature for detection, therapeutic monitoring, and prognosis in osteosarcoma

Abstract

Osteosarcoma (OS) is the primary bone tumor in children and young adults. Currently, there are no reliable, noninvasive biologic markers to detect the presence or progression of disease, assess therapy response, or provide upfront prognostic insights. MicroRNAs (miRNAs) are evolutionarily conserved, stable, small noncoding RNA molecules that are key posttranscriptional regulators and are ideal candidates for circulating biomarker development due to their stability in plasma, ease of isolation, and the unique expressions associated with specific disease states. Using a qPCR-based platform that analyzes more than 750 miRNAs, we analyzed control and diseased-associated plasma from a genetically engineered mouse model of OS to identify a profile of four plasma miRNAs. Subsequent analysis of 40 human patient samples corroborated these results. We also identified disease-specific endogenous reference plasma miRNAs for mouse and human studies. Specifically, we observed plasma miR-205-5p was decreased 2.68-fold in mice with OS compared to control mice, whereas, miR-214, and miR-335-5p were increased 2.37- and 2.69-fold, respectively. In human samples, the same profile was seen with miR-205-5p decreased 1.75-fold in patients with OS, whereas miR-574-3p, miR-214, and miR-335-5p were increased 3.16-, 8.31- and 2.52-fold, respectively, compared to healthy controls. Furthermore, low plasma levels of miR-214 in metastatic patients at time of diagnosis conveyed a significantly better overall survival. This is the first study to identify plasma miRNAs that could be used to prospectively identify disease, potentially monitor therapeutic efficacy and have prognostic implications for OS patients.

miR‑214‑3p promotes the proliferation, migration and invasion of osteosarcoma cells by targeting CADM1

“…Previous studies have demonstrated that miR-214 functions either as an oncogene or a tumor suppressor in a number of human cancer types, including lung, prostate, colorectal and esophageal cancer (15)(16)(17)(18). Previous studies have indicated that elevated miR-214-3p is associated with OS progression, but a limited number of studies have focused on the function and mechanism of miR-214-3p (10,11,19,20). In a previous study by the authors, it was demonstrated that upregulated miR-214 expression was associated with aggressive clinicopathological features (tumor size, metastasis status and response to pre-operative chemotherapy) and poor prognosis of pediatric osteosarcoma (11).…”

“…This suggests that miR-214 may function as a tumor suppressor and may have a tumorigenic role. In previous studies, researchers have demonstrated that miR-214 was upregulated in osteosarcoma and associated with tumor progression and poor prognosis (10,11). However, little progress has been achieved in terms of elucidating the molecular mechanisms of miR-214-3p-mediated tumorigenesis in OS.…”

Role of miRNA‐542‐5p in the tumorigenesis of osteosarcoma

“…GEO (http://www.ncbi.nlm.nih.gov/geo) is a public functional genomics data repository for high‐throughput gene expression data, different types of microarrays and chips [9]. The miRNA microarray datasets (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE69470 [10], http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE28425 [11] and http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE65071 [12]) were downloaded from GEO (GPL20275, GPL8227 and GPL19631 platform, respectively), and we performed an integrated analysis. Comprehensive miRNA expression profiling was obtained by using the probes in the platforms mentioned earlier.…”

Toward the promise of microRNAs – Enhancing reproducibility and rigor in microRNA research

“…42 If the experiment is using an array-based method with hundreds of microRNAs, a global normalization approach may be useful. 33,43 Many microRNAs Figure 3. microRNA-seq normalization (RPM) is a dependent normalization.…”

“…AllenRhoades et al designed a study to identify plasma microRNAs that could detect and monitor childhood osteosarcoma. 43 The first component was performed in an osteosarcoma mouse model. After power calculations were performed on how many animals would be required, human osteosarcoma tumors were grown in nude mice with serial plasma collections over time.…”