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Serum microRNA array analysis identifies miR-140-3p, miR-33b-3p and miR-671-3p as potential osteoarthritis biomarkers involved in metabolic processes.
Clin Epigenetics. 2017; 9:127.CE

Abstract

BACKGROUND

MicroRNAs (miRNAs) in circulation have emerged as promising biomarkers. In this study, we aimed to identify a circulating miRNA signature for osteoarthritis (OA) patients and in combination with bioinformatics analysis to evaluate the utility of selected differentially expressed miRNAs in the serum as potential OA biomarkers.

METHODS

Serum samples were collected from 12 primary OA patients, and 12 healthy individuals were screened using the Agilent Human miRNA Microarray platform interrogating 2549 miRNAs. Receiver Operating Characteristic (ROC) curves were constructed to evaluate the diagnostic performance of the deregulated miRNAs. Expression levels of selected miRNAs were validated by quantitative real-time PCR (qRT-PCR) in all serum and in articular cartilage samples from OA patients (n = 12) and healthy individuals (n = 7). Bioinformatics analysis was used to investigate the involved pathways and target genes for the above miRNAs.

RESULTS

We identified 279 differentially expressed miRNAs in the serum of OA patients compared to controls. Two hundred and five miRNAs (73.5%) were upregulated and 74 (26.5%) downregulated. ROC analysis revealed that 77 miRNAs had area under the curve (AUC) > 0.8 and p < 0.05. Bioinformatics analysis in the 77 miRNAs revealed that their target genes were involved in multiple signaling pathways associated with OA, among which FoxO, mTOR, Wnt, pI3K/akt, TGF-β signaling pathways, ECM-receptor interaction, and fatty acid biosynthesis. qRT-PCR validation in seven selected out of the 77 miRNAs revealed 3 significantly downregulated miRNAs (hsa-miR-33b-3p, hsa-miR-671-3p, and hsa-miR-140-3p) in the serum of OA patients, which were in silico predicted to be enriched in pathways involved in metabolic processes. Target-gene analysis of hsa-miR-140-3p, hsa-miR-33b-3p, and hsa-miR-671-3p revealed that InsR and IGFR1 were common targets of all three miRNAs, highlighting their involvement in regulation of metabolic processes that contribute to OA pathology. Hsa-miR-140-3p and hsa-miR-671-3p expression levels were consistently downregulated in articular cartilage of OA patients compared to healthy individuals.

CONCLUSIONS

A serum miRNA signature was established for the first time using high density resolution miR-arrays in OA patients. We identified a three-miRNA signature, hsa-miR-140-3p, hsa-miR-671-3p, and hsa-miR-33b-3p, in the serum of OA patients, predicted to regulate metabolic processes, which could serve as a potential biomarker for the evaluation of OA risk and progression.

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Authors+Show Affiliations

Ntoumou E
Laboratory of Cytogenetics and Molecular Genetics, Faculty of Medicine, Biopolis, University of Thessaly, 41500 Larissa, Greece.
Tzetis M
Laboratory of Medical Genetics, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
Braoudaki M
Laboratory of Medical Genetics, Medical School, National and Kapodistrian University of Athens, Athens, Greece. First Department of Pediatrics, Medical School, National and Kapodistrian University of Athens,, Athens, Greece. University Research Institute for the Study and Treatment of Childhood Disease and Malignant Diseases, National and Kapodistrian University of Athens, "Aghia Sophia" Children's Hospital, Athens, Greece.
Lambrou G
First Department of Pediatrics, Medical School, National and Kapodistrian University of Athens,, Athens, Greece. University Research Institute for the Study and Treatment of Childhood Disease and Malignant Diseases, National and Kapodistrian University of Athens, "Aghia Sophia" Children's Hospital, Athens, Greece.
Poulou M
Laboratory of Medical Genetics, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
Malizos K
Department of Orthopaedics, Faculty of Medicine, University of Thessaly, Larissa, Greece.
Stefanou N
Department of Orthopaedics, Faculty of Medicine, University of Thessaly, Larissa, Greece.
Anastasopoulou L
Laboratory of Cytogenetics and Molecular Genetics, Faculty of Medicine, Biopolis, University of Thessaly, 41500 Larissa, Greece.
Tsezou A
Laboratory of Cytogenetics and Molecular Genetics, Faculty of Medicine, Biopolis, University of Thessaly, 41500 Larissa, Greece. Department of Biology, Faculty of Medicine, University of Thessaly, Larissa, Greece.

MeSH

AgedComputational BiologyDown-RegulationFemaleGene Expression ProfilingGene Expression Regulation, NeoplasticGene Regulatory NetworksGenetic MarkersHumansMaleMicroRNAsMiddle AgedOligonucleotide Array Sequence AnalysisOsteoarthritisSensitivity and Specificity

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29255496

Citation

Ntoumou, E, et al. "Serum microRNA Array Analysis Identifies miR-140-3p, miR-33b-3p and miR-671-3p as Potential Osteoarthritis Biomarkers Involved in Metabolic Processes." Clinical Epigenetics, vol. 9, 2017, p. 127.
Ntoumou E, Tzetis M, Braoudaki M, et al. Serum microRNA array analysis identifies miR-140-3p, miR-33b-3p and miR-671-3p as potential osteoarthritis biomarkers involved in metabolic processes. Clin Epigenetics. 2017;9:127.
Ntoumou, E., Tzetis, M., Braoudaki, M., Lambrou, G., Poulou, M., Malizos, K., Stefanou, N., Anastasopoulou, L., & Tsezou, A. (2017). Serum microRNA array analysis identifies miR-140-3p, miR-33b-3p and miR-671-3p as potential osteoarthritis biomarkers involved in metabolic processes. Clinical Epigenetics, 9, 127. https://doi.org/10.1186/s13148-017-0428-1
Ntoumou E, et al. Serum microRNA Array Analysis Identifies miR-140-3p, miR-33b-3p and miR-671-3p as Potential Osteoarthritis Biomarkers Involved in Metabolic Processes. Clin Epigenetics. 2017;9:127. PubMed PMID: 29255496.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Serum microRNA array analysis identifies miR-140-3p, miR-33b-3p and miR-671-3p as potential osteoarthritis biomarkers involved in metabolic processes. AU - Ntoumou,E, AU - Tzetis,M, AU - Braoudaki,M, AU - Lambrou,G, AU - Poulou,M, AU - Malizos,K, AU - Stefanou,N, AU - Anastasopoulou,L, AU - Tsezou,A, Y1 - 2017/12/12/ PY - 2017/8/21/received PY - 2017/11/29/accepted PY - 2017/12/20/entrez PY - 2017/12/20/pubmed PY - 2018/9/27/medline KW - Biomarker KW - Circulating miRNAs KW - Hsa-miR- 671-3p KW - Hsa-miR-140-3p KW - Hsa-miR-33b-3p KW - Metabolic KW - Osteoarthritis KW - miR-array SP - 127 EP - 127 JF - Clinical epigenetics JO - Clin Epigenetics VL - 9 N2 - BACKGROUND: MicroRNAs (miRNAs) in circulation have emerged as promising biomarkers. In this study, we aimed to identify a circulating miRNA signature for osteoarthritis (OA) patients and in combination with bioinformatics analysis to evaluate the utility of selected differentially expressed miRNAs in the serum as potential OA biomarkers. METHODS: Serum samples were collected from 12 primary OA patients, and 12 healthy individuals were screened using the Agilent Human miRNA Microarray platform interrogating 2549 miRNAs. Receiver Operating Characteristic (ROC) curves were constructed to evaluate the diagnostic performance of the deregulated miRNAs. Expression levels of selected miRNAs were validated by quantitative real-time PCR (qRT-PCR) in all serum and in articular cartilage samples from OA patients (n = 12) and healthy individuals (n = 7). Bioinformatics analysis was used to investigate the involved pathways and target genes for the above miRNAs. RESULTS: We identified 279 differentially expressed miRNAs in the serum of OA patients compared to controls. Two hundred and five miRNAs (73.5%) were upregulated and 74 (26.5%) downregulated. ROC analysis revealed that 77 miRNAs had area under the curve (AUC) > 0.8 and p < 0.05. Bioinformatics analysis in the 77 miRNAs revealed that their target genes were involved in multiple signaling pathways associated with OA, among which FoxO, mTOR, Wnt, pI3K/akt, TGF-β signaling pathways, ECM-receptor interaction, and fatty acid biosynthesis. qRT-PCR validation in seven selected out of the 77 miRNAs revealed 3 significantly downregulated miRNAs (hsa-miR-33b-3p, hsa-miR-671-3p, and hsa-miR-140-3p) in the serum of OA patients, which were in silico predicted to be enriched in pathways involved in metabolic processes. Target-gene analysis of hsa-miR-140-3p, hsa-miR-33b-3p, and hsa-miR-671-3p revealed that InsR and IGFR1 were common targets of all three miRNAs, highlighting their involvement in regulation of metabolic processes that contribute to OA pathology. Hsa-miR-140-3p and hsa-miR-671-3p expression levels were consistently downregulated in articular cartilage of OA patients compared to healthy individuals. CONCLUSIONS: A serum miRNA signature was established for the first time using high density resolution miR-arrays in OA patients. We identified a three-miRNA signature, hsa-miR-140-3p, hsa-miR-671-3p, and hsa-miR-33b-3p, in the serum of OA patients, predicted to regulate metabolic processes, which could serve as a potential biomarker for the evaluation of OA risk and progression. SN - 1868-7083 UR - https://www.unboundmedicine.com/medline/citation/29255496/Serum_microRNA_array_analysis_identifies_miR_140_3p_miR_33b_3p_and_miR_671_3p_as_potential_osteoarthritis_biomarkers_involved_in_metabolic_processes_ DB - PRIME DP - Unbound Medicine ER -