ECTS2016 Poster Presentations Genetics and Epigenetics (25 abstracts)
Identification and functional validation of microRNA expression in human bone tissue
Barbara Ostanek 1 , Simona Mencej Bedrac 1 , Barbara Kern 1 , Peter Vrtacnik 1 , Vid Mlakar 1 , Tamer Bego 2 , Radko Komadina 3 , Rihard Trebše 4 & Janja Marc 1
1Faculty of Pharmacy, Department of Clinical Biochemistry, University of Ljubljana, Ljubljana, Slovenia; 2Faculty of Pharmacy, Department of Clinical Biochemistry, University of Sarajevo, Sarajevo, Bosnia and Herzegovina; 3General Hospital Celje, Department of Traumatology, Celje, Slovenia; 4Orthopedics Hospital Valdoltra, Valdoltra, Slovenia.
miRNAs are epigenetic regulators of gene expression, increasingly recognised as prominent regulators of bone metabolism. Following our in vitro studies of miRNAs profiles in osteoblasts we extended the research to human bone samples. Our aim was to find and functionally validate miRNAs differentially expressed in osteoporotic and osteoarthrotic bone with potential for targeting genes relevant in bone metabolism.
Trabecular bone tissue samples were collected from 44 osteoporotic patients (OP) with hip fracture and 44 patients with primary osteoarthrosis (OA) of the hip during arthroplastic surgery following ethical approval. Fifteen autopsy cases served as a control group (C). In the discovery stage Nanostring nCounter technology was used to obtain the expression profile of 800 miRNAs from 5 OP and 5 OA bone samples. qPCR was used for the validation of differentially expressed miRNAs and their predicted targets.
Nine miRNAs were differentially expressed in OP and OA groups in the discovery stage. miR-195-5p and miR-204-5p were selected for validation in the whole group of 103 patients. Additionally, their potential target genes relevant in bone metabolism were identified using three target prediction tools and were measured. There was a significant difference in miR-195-5p but not miR-204-5p levels among the tested groups (P<0.0001). Furthermore, there was a negative correlation between the levels of miR-195-5p and possible target genes ELOVL6 (P=0.001), GSTCD (P<0.0001) and MYB (P=0.035). Their gene expressions were also significantly different in the three groups with the highest levels in OP followed by OA and C groups (P<0.0001 for all three genes).
Our results are the first to identify miR-195-5p as a potential biomarker in OP and OA, which has only recently been demonstrated to inhibit osteoblast differentiation. Moreover, negative correlations with ELOVL6, GSTCD and MYB gene expression represent evidence of interaction. Further functional confirmations of predicted target genes are underway.
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Bone Abstracts
ISSN 2052-1219 (online)
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