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Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources

  • Authors:
    • Hui Zhang
    • Shiwei Wu
    • Dan Luo
    • Jincai Guo
  • View Affiliations / Copyright

    Affiliations: Department of Pharmacy, Changsha Stomatological Hospital, Changsha, Hunan 410006, P.R. China, Department of Clinical Pharmacy, Xiangtan Center Hospital, The Affiliated Hospital of Hunan University, Xiangtan, Hunan 411100, P.R. China, Department of Endodontics, Changsha Stomatological Hospital, Changsha, Hunan 410006, P.R. China, Department of Pharmacy, Changsha Stomatological Hospital, Changsha, Hunan 410006, P.R. China
    Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 263
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    Published online on: April 24, 2026
       https://doi.org/10.3892/ol.2026.15618
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Abstract

Osteoclast differentiation and activation pathways drive abnormally enhanced bone resorption in osteosarcoma, and have thus emerged as potential therapeutic targets for this malignancy. However, the diagnostic value of osteoclast differentiation‑related genes (ODRGs) in osteosarcoma remains largely uncharacterized. The present study first analyzed the expression profiles of ODRGs in osteosarcoma using multiple Gene Expression Omnibus datasets. Analysis of single cell RNA‑sequencing data from osteosarcoma tissues demonstrated that colony‑stimulating factor‑1 receptor (CSF1R), a key ODRG, was widely expressed in osteoblasts and monocyte/macrophage lineages within the osteosarcoma microenvironment. Subgroup analysis further revealed that patients with osteosarcoma in the low‑CSF1R expression group exhibited significantly increased sensitivity to immune checkpoint inhibitors compared with those in the high‑CSF1R expression group. Furthermore, pan‑cancer analysis across The Cancer Genome Atlas datasets demonstrated that CSF1R expression was aberrantly regulated in multiple tumor types and strongly correlated with the expression levels of immune checkpoint markers and the infiltration levels of immune cells. Functional validation experiments confirmed that treatment with CSF1R‑specific inhibitors significantly reduced the viability of the MG63 and Saos‑2 cell lines. To identify novel, natural‑product‑derived CSF1R inhibitors, structure‑based virtual screening was combined with in vitro experimental assays (CCK‑8 and western blotting). Among the validated candidates, sarsasapogenin, a compound identified from the screen, effectively suppressed CSF1R protein expression and inhibited the proliferation of both MG63 and Saos‑2 cells in a dose‑dependent manner. Collectively, the present findings highlight CSF1R as a clinically promising diagnostic biomarker and therapeutic target for osteosarcoma.
View Figures

Figure 1

Identification of ODRGs in
osteosarcoma. Volcano plots depicting the DEGs in the (A)
GEO-GSE33382, (B) GEO-GSE14359 and (C) GEO-GSE218035 datasets (log2
FC >1; P<0.05). (D) A Venn diagram identifying 156 DEGs in
the overlap of the three datasets retrieved from the GEO database.
(E) KEGG enrichment analysis of the 156 DEGs. (F) WIKI pathways
enrichment analysis of the 156 DEGs. (G) Analysis of expression
levels of osteoclast differentiation genes (FHL2, ACP5, FOSL1,
CSF1R and TNFRSF11B) in the GSE33382 datasets using an unpaired
two-tailed Student's t-test. (H) The expression levels of
osteoclast differentiation genes were assessed between osteosarcoma
tissues and normal bone tissues (n=2) in the GSE14359 dataset. (I)
Analysis of expression levels of osteoclast differentiation genes
(FHL2, ACP5, FOSL1, CSF1R and TNFRSF11B) in the GSE218035 datasets
using an unpaired two-tailed Student's t-test. *P<0.05,
**P<0.01, ***P<0.001 and ****P<0.0001. DEGs,
differentially expressed genes; KEGG, Kyoto Encyclopedia of Genes
and Genomes; CSF1R, colony-stimulating factor-1 receptor; FHL2,
four and a half LIM domains 2; ACP5, acid phosphatase 5, tartrate
resistant; FOSL1, FOS-like 1, AP-1 transcription factor subunit;
TNFRSF11B, TNF receptor superfamily member 11b.

Figure 2

Assessment of the clinical response
to immune checkpoint inhibitor therapy. (A) In the gene chip array
cohort of osteosarcoma, differences in TIDE algorithm scores of
CSF1R between the high- and low-expression groups in the TARGET
dataset were assessed (Wilcoxon rank sum test). (B) In the RNA-seq
data of osteosarcoma, differences in TIDE algorithm scores of CSF1R
between the high- and low-expression groups in the TARGET dataset
were assessed (Wilcoxon rank sum test). (C) The sensitivity of
immune checkpoint between the high- and low-expression groups of
CSF1R in the TARGET dataset (gene chip array data in osteosarcoma).
(D) The sensitivity of immune checkpoint between the high- and
low-expression groups of CSF1R in the TARGET dataset (RNA-seq data
in osteosarcoma). **P<0.01 and ***P<0.001. ns, not
significant; CSF1R, colony-stimulating factor-1 receptor; G1,
high-expression group; G2, low-expression group; RNA-seq,
Ribonucleic Acid-sequencing; TARGET, Therapeutically Applicable
Research To Generate Effective Treatments; TIDE, Tumor Immune
Dysfunction and Exclusion.

Figure 3

Single-cell sequencing analysis of
CSF1R. (A) UMAP plot showing annotated cell categories after
dimension reduction in the GSE162454 dataset. (B) UMAP plot showing
the expression distribution of CSF1R in different cell categories.
(C) Violin plot showing the results of CSF1R expression
distribution in different cell categories from UMAP data analysis.
(D) Violin plot representing relative expression levels of CSF1R
for different cell types in 6 patients with osteosarcoma. CD4Tconx,
CD4+ conventional T cells; CD8Tex, CD8+ exhausted T cells;
mono/macro, monocytes/macrophages; CSF1R, colony-stimulating factor
1 receptor; N.S., not significant.

Figure 4

Cell viability (%) of MG63 and Saos-2
cells treated with pexidartinib (via PLX3397) and sotuletinib.

Figure 5

Sensitization of the MG63 and Saos-2
cell lines to a CSF1R inhibitor. (A) Dose-dependent cytotoxic
effects of linarin on MG63 and Saos-2 osteosarcoma cell viability.
(B) Dose-dependent cytotoxic effects of saikosaponin C on MG63 and
Saos-2 osteosarcoma cell viability. (C) Dose-dependent cytotoxic
effects of sarsasapogenin on MG63 and Saos-2 osteosarcoma cell
viability. (D) Molecular docking of the binding between
sarsasapogenin and the CSF1R protein. (E) Visual molecular docking
of the binding acid residues between sarsasapogenin and CSF1R
protein. (F) The expression levels of CSF1R as detected by western
blot analysis in MG63 and Saos-2 cells after treatment with
sarsasapogenin. *P<0.05. CSF1R, colony-stimulating factor-1
receptor.

Figure 6

Molecular dynamics simulation
analyses. (A) RMSD curves of sarsasapogenin with the CSF1R protein.
(B) The Rg analysis of sarsasapogenin with the CSF1R protein. (C)
RMSF curves of sarsasapogenin with the CSF1R protein. (D) The SASA
of sarsasapogenin with CSF1R protein. (E) H-bond number of
sarsasapogenin with the CSF1R protein. CSF1R, colony-stimulating
factor-1 receptor; RMSD, root mean square deviation; Rg, radius of
gyration; RMSF, root mean square fluctuation; SASA,
solvent-accessible surface area.

Figure 7

Relative binding free energies of
sarsasapogenin-CSF1R complexes. (A) Binding free energy component
analysis of sarsasapogenin with the CSF1R protein. (B) Key amino
acid energy analysis of sarsasapogenin with the CSF1R protein.
CSF1R, colony-stimulating factor-1 receptor. VDWAALs, Van der
Waals; MM/PBSA, molecular mechanics Poisson-Boltzmann surface
area.
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Zhang H, Wu S, Luo D and Guo J: Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources. Oncol Lett 31: 263, 2026.
APA
Zhang, H., Wu, S., Luo, D., & Guo, J. (2026). Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources. Oncology Letters, 31, 263. https://doi.org/10.3892/ol.2026.15618
MLA
Zhang, H., Wu, S., Luo, D., Guo, J."Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources". Oncology Letters 31.6 (2026): 263.
Chicago
Zhang, H., Wu, S., Luo, D., Guo, J."Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources". Oncology Letters 31, no. 6 (2026): 263. https://doi.org/10.3892/ol.2026.15618
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Spandidos Publications style
Zhang H, Wu S, Luo D and Guo J: Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources. Oncol Lett 31: 263, 2026.
APA
Zhang, H., Wu, S., Luo, D., & Guo, J. (2026). Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources. Oncology Letters, 31, 263. https://doi.org/10.3892/ol.2026.15618
MLA
Zhang, H., Wu, S., Luo, D., Guo, J."Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources". Oncology Letters 31.6 (2026): 263.
Chicago
Zhang, H., Wu, S., Luo, D., Guo, J."Computational discovery of novel colony‑stimulating factor‑1 receptor as a potential therapeutic biomarker in osteosarcoma and a novel inhibitor from herbal sources". Oncology Letters 31, no. 6 (2026): 263. https://doi.org/10.3892/ol.2026.15618
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