microRNA-25-3p suppresses osteogenic differentiation of BMSCs in patients with osteoporosis by targeting ITGB3
Introduction
Osteoporosis (OP) is the most common disease of bone metabolism with a very high prevalence, and women are usually at higher risk than men (Ensrud and Crandall, 2017). OP is typified by low bone mass and microarchitectural degeneration of bone tissues, leading to increased bone frangibility and subsequently elevated danger of fracture (1993). Age is one of the most important factors influencing the incidence of fractures caused by OP (Ensrud and Crandall, 2017). OP and fractures associated with OP are common causes of morbidity and mortality in the elderly (Johnston and Dagar, 2020). OP is also considered a silent disease because there is usually no symptom until the first fracture occurs (Johnston and Dagar, 2020). In fact, OP is caused by an imbalance between bone resorption by osteoclasts and bone formation by osteoblasts (Wang et al., 2021). The most basic and effective measure to treat OP is to reduce bone resorption and increase bone formation (Wang et al., 2021). It has been proposed that microRNAs (miRNAs) affect the osteogenic differentiation process of bone marrow mesenchymal stem cells (BMSCs) (Wang et al., 2019).
miRNAs, a class of small non-coding RNAs, can bind to target genes to participate in various biological processes, including cell propagation, differentiation, and apoptosis, as well as organ development (Wang et al., 2021). Liu et al. identified miR-25–3p as a repressor of osteogenesis in human umbilical cord stem cells (Cao et al., 2020). miR-25–3p is a suitable reference gene for OP induced by estrogen deficiency and mechanical unloading because miR-25–3p was stably expressed in OP or non-OP serum and bone tissues (Chen et al., 2016). On contrary, evidence in a previous study also showed that miR-25–3p regulated the expression of NFIX to facilitate osteoclast activity (Huang et al., 2020). Considering the controversial role of miR-25–3p in osteogenic differentiation, this study aims to determine whether miR-25–3p has a certain role in osteogenic differentiation of BMSCs.
A study has suggested that integrin-beta3 (ITGB3) is an osteoblast-specific gene (Asagiri and Takayanagi, 2007). It has been demonstrated that IL1RN interacts with ITGB3 to activate β-linked protein signaling, which ultimately promotes osteoblast differentiation (Zou et al., 2021), indicating the possible implication of ITGB3 in osteoblast differentiation. The binding sites between miR-25–3p and ITGB3 were predicted by starBase. We, therefore, determined the possible effect of the miR-25–3p/ITGB3 axis on BMSC osteogenic differentiation under the context of OP.
Section snippets
Sample collection
Bone marrow tissues were collected from six patients [aged (53 ± 6.92) years, including 4 females and 2 males] diagnosed with OP who underwent hip surgery at the First Hospital of Changsha, and normal bone marrow tissues donated by three injured patients [aged (50 ± 4.85) years, including 2 females and 1 male] were also harvested. Patients with gout, rheumatoid arthritis, systemic inflammatory diseases, autoimmune diseases, or chronic malignant diseases were excluded from this study. Written
Osteogenic differentiation of BMSCs was reduced in OP patients
To identify whether cells isolated from bone marrow were BMSCs, the expression of BMSC biomarkers (CD73, CD90, CD105, CD34, and CD45) was measured by flow cytometry. CD73, CD90, and CD105 were utilized as positive markers for BMSCs, and the hematopoietic stem cell marker CD34 and the leukocyte marker CD45 were used as negative markers. The results showed that the cells separated from bone marrow of normal controls (normal-BMSCs) and OP patients (OP-BMSCs) had the positive expression of CD73,
Discussion
OP is a common disease that causes severe bone loss and may lead to pathological fractures, which can reduce activities of daily living and quality of life. Therefore, OP prevention needs to be taken seriously. In our research, miR-25–3p and ITGB3 expression was determined in BMSCs extracted from OP patients and normal individuals. The findings indicated that ITGB3 was lowly expressed and miR-25–3p was highly expressed in OP-BMSCs. Moreover, this study further demonstrated that miR-25–3p
Conclusion
In summary, our findings stated that the miR-25–3p/ITGB3 axis regulated osteogenic differentiation of BMSCs from OP patients. This study aims to provide some new theoretical and experimental basis for OP treatment. The limitation of this study is that only the impact of the miR-25–3p/ITGB3 axis on BMSC osteogenic differentiation was investigated. The upstream and downstream of this axis will be further investigated in the future to advance our understanding of the miR-25–3p/ITGB3 axis in BMSC
Author statement
No conflict of interest exits in the submission of this manuscript, and manuscript is approved by all authors for publication. I would like to declare on behalf of my co-authors that the work described was original research that has not been published previously, and not under consideration for publication elsewhere, in whole or in part. All the authors listed have approved the manuscript that is enclosed.
Funding
This study was supported by Projects of Health and Family Planning Commission of Jiangxi Provincial (No.20177013; No.202140022), Changsha Science and Technology Bureau (No.kq2004157) and Hunan Provincial Health Commission (No.202201042974).
Authors’ contributions
YDP conceived the ideas. YDP and LZ designed the experiments. YDP, LZ and CJ performed the experiments. YDP and SF analyzed the data. YDP, LZ and WGW provided critical materials. YDP and LZ wrote the manuscript. YDP supervised the study. All the authors have read and approved the final version for publication.
Competing interest
The authors declare there is no conflict of interest regarding the publication of this paper.
Acknowledgment
We acknowledge Guowen Wei (Department of Orthopedics, The First Hospital of Nanchang, Nanchang, China) for her valuable advice. We acknowledge Zhen Li (Department of Pathology, The First Hospital of Changsha City, Changsha, China) for her support.
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Dongping Yu and Zhen Li contributed equally to this research.