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Updated: Mar 18, 2026
Differentiation of Functional Osteoclasts from Human Peripheral Blood CD14+ Monocytes
Published on: January 27, 2023
5α-dihydrotestosterone upregulates interferon signaling and limits osteoclast differentiation during inflammatory
Kiana Chen1,2, Michael Christof2, Gulzada Kulzhanova2
1Department of Pathology & Laboratory Medicine, University of Rochester Medical Center, Rochester, NY, USA.
Purpose:
Rheumatoid arthritis is characterized by inflammatory erosions and increased prevalence in females. Androgen, the dominant sex hormone in males, is protective against bone loss. Here, we investigated the cellular targets of androgen and hypothesized that androgen negatively regulates TNFα-driven osteoclastogenesis.
Materials And Methods:
Bone marrow-derived cells from a C57BL/6J mouse femur and tibia were plated with 10-9M of 5α-dihydrotestosterone (DHT) or vehicle then collected for flow cytometry and flow sorting. Osteoclast precursors (OCPs), myeloid-like CD11b+Gr-1- cells, were grown to osteoclasts then fixed and TRAP-stained. Osteoclasts were quantified by cell counting. Single-cell RNA-sequencing was performed on DHT or vehicle-treated TNFα-stimulated cells, and the differences in cell populations, gene expression, gene ontology, and cell trajectory were analyzed via bioinformatic approaches. RT-qPCR validated sequencing results.
Results:
DHT-treated OCPs were decreased versus vehicle-treated OCPs (Vehicle = 12.43 ± 0.53%, DHT = 9.93 ± 0.85%, p < 0.05), and there was decreased differentiation from OCPs to osteoclasts (Vehicle = 403.3 ± 141.5, DHT = 10.67 ± 14.36, p < 0.01). Analysis identified osteoclastogenesis-related clusters that varied between samples and interferon-related processes upregulated with DHT treatment. Analysis also determined that DHT treatment may be associated with expression of the transcription factor Irf7. Isg15, the most differentially expressed gene in the DHT-treated clusters, may also be involved in osteoclastogenesis.
Conclusions:
In summary, androgen targets OCPs and limits their differentiation into osteoclasts. In myeloid populations, DHT treatment results in differential expression of interferon-related processes and genes and alters the trajectory of cells, potentially regulating osteoclastogenesis.
Insights
Androgen, like 5α-dihydrotestosterone (DHT), inhibits osteoclast formation from osteoclast precursors (OCPs). This study reveals DHT alters myeloid cell gene expression, potentially regulating inflammatory bone erosion in rheumatoid arthritis.
Area of Science:
- Endocrinology
- Immunology
- Rheumatology
Background:
- Rheumatoid arthritis (RA) involves inflammatory bone erosions, with higher prevalence in females.
- Androgens, like testosterone, are protective against bone loss and dominant in males.
- Tumor necrosis factor-alpha (TNFα) drives osteoclastogenesis, a key process in RA bone destruction.
Purpose of the Study:
- To identify the cellular targets of androgen action.
- To investigate the hypothesis that androgens negatively regulate TNFα-induced osteoclastogenesis.
Main Methods:
- Bone marrow-derived cells from mice were treated with 5α-dihydrotestosterone (DHT) or vehicle.
- Osteoclast precursors (OCPs) were isolated, cultured, and differentiated into osteoclasts.
- Single-cell RNA sequencing and RT-qPCR were used to analyze gene expression and cell populations.
Main Results:
- DHT treatment decreased the number of OCPs and their differentiation into osteoclasts.
- DHT altered gene expression in myeloid cells, upregulating interferon-related pathways.
- DHT treatment was associated with the expression of transcription factor Irf7 and the gene Isg15.
Conclusions:
- Androgens target OCPs, limiting their differentiation into osteoclasts.
- DHT influences myeloid cell gene expression and cell trajectory, potentially regulating osteoclastogenesis.
- These findings suggest a role for androgens in modulating inflammatory bone loss relevant to rheumatoid arthritis.
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