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Two Techniques to Create Hypoparathyroid Mice: Parathyroidectomy Using GFP Glands and Diphtheria-Toxin-Mediated Parathyroid Ablation
Published on: March 14, 2017
MicroRNA-874-3p is a Potential Contributor to Primary Hyperparathyroidism-Induced Osteoporosis
Kaiyuan Cheng1, Ruifeng Bai2, Minjuan Li1
1Department of Orthopedic Trauma, Beijing Jishuitan Hospital, Capital Medical University, Beijing, People's Republic of China.
Background:
Dysregulation of microRNAs contributes to bone diseases. However, the microRNAs involved in primary hyperparathyroidism (PHPT)-induced osteoporosis remain unknown.
Methods:
The parathyroid tissue samples were obtained from PHPT patients with or without osteoporosis (n = 5/group) during parathyroid resection and subjected to high throughput microRNA sequencing. The differentially expressed microRNAs were identified and further verified using qRT-PCR. Alizarin Red Staining was performed to detected the osteogenic differentiation. Gain- and loss-of-function assays were performed to investigate the role of miR-874-3p, which was upregulated in PHPT patients with osteoporosis, in human mesenchymal stem cells (hMSCs) undergoing osteoblastic differentiation.
Results:
We identified 32 significantly upregulated and 18 significantly downregulated microRNAs in PHPT patients with osteoporosis. miR-874-3p was increased in PHPT osteoporosis patients, meanwhile, miR-874-3p in parathyroid tissue and peripheral blood extracellular vesicles of PHPT osteoporosis mice were increased. The miR-874-3p level was remarkably elevated in hMSCs grown in osteogenic medium. Overexpression of miR-874-3p repressed the hMSC osteogenic differentiation and reduced the osteogenic marker expression in hMSCs, whereas miR-874-3p inhibitor showed a contrasting effect. The results of the dual luciferase reporting system showed that miR-874-3p could reduce the luciferase activity of wild-type FTO-WT-3 '-UTR. However, there was no significant change in the luciferase activity of the mutant compared with the control group.
Conclusion:
MiR-874-3p might specifically binds to FTO suppress osteogenic differentiation of hMSCs, thereby contributing to the development of osteoporosis in PHPT patients.
Insights
MicroRNAs impact bone health. This study identifies miR-874-3p as a key microRNA involved in primary hyperparathyroidism-induced osteoporosis by targeting FTO and suppressing bone formation.
Area of Science:
- Endocrinology
- Molecular Biology
- Bone Biology
Background:
- MicroRNA dysregulation is implicated in various bone diseases.
- The specific microRNAs contributing to primary hyperparathyroidism (PHPT)-induced osteoporosis are not well understood.
Purpose of the Study:
- To identify microRNAs involved in PHPT-induced osteoporosis.
- To investigate the role of miR-874-3p in the osteogenic differentiation of human mesenchymal stem cells (hMSCs).
Main Methods:
- High-throughput microRNA sequencing of parathyroid tissue from PHPT patients with or without osteoporosis.
- Quantitative real-time PCR (qRT-PCR) for microRNA verification.
- Gain- and loss-of-function assays in hMSCs to assess miR-874-3p's role in osteogenic differentiation.
- Dual luciferase reporter assays to confirm target interaction.
Main Results:
- Identified 32 upregulated and 18 downregulated microRNAs in PHPT osteoporosis patients.
- miR-874-3p was significantly upregulated in PHPT osteoporosis patients, parathyroid tissue, and peripheral blood extracellular vesicles of PHPT osteoporosis mice.
- Overexpression of miR-874-3p inhibited hMSC osteogenic differentiation and reduced osteogenic marker expression, while inhibition had the opposite effect.
- miR-874-3p was confirmed to bind to FTO.
Conclusions:
- miR-874-3p is upregulated in PHPT patients with osteoporosis.
- miR-874-3p suppresses osteogenic differentiation of hMSCs by targeting FTO.
- miR-874-3p plays a crucial role in the pathogenesis of PHPT-induced osteoporosis.
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